HK40006110A

HK40006110A - Methods and compositions for treating non-erk mapk pathway inhibitor-resistant cancers

Info

Publication number: HK40006110A
Application number: HK19129653.2A
Authority: HK
Inventors: S‧萨哈; D‧威尔施; G‧德克雷斯森佐; J‧J‧罗伊克斯
Original assignee: 生物医学谷探索股份有限公司
Priority date: 2016-05-20
Filing date: 2017-05-22
Publication date: 2020-05-15

Description

Methods and compositions for treating non-ERK MAPK pathway inhibitor resistant cancers

Cross Reference to Related Applications

This application claims the benefit of U.S. patent application serial No. 15/161,137 filed on month 5 and 20 of 2016, which is a continuation-in-part application of PCT international application number PCT/US2014/071749 filed on month 12 and 19 of 2014, where PCT international application number PCT/US2014/071749 claims the benefit of U.S. patent application serial No. 61/919,551 filed on month 12 and 20 of 2013, all of which are incorporated herein by reference in their entirety as if fully set forth herein.

Technical Field

The invention provides, inter alia, methods, pharmaceutical compositions, and kits for treating or ameliorating the effects of a cancer in a subject that is refractory or resistant to non-ERK MAPK pathway inhibitor therapy.

Incorporation by reference of sequence listing

This application contains references to amino acid and/or nucleic acid sequences that have been filed concurrently herewith as sequence listing text file "0398850 pct.txt" with file size 351KB created on day 5/20 of 2016. The above sequence listing is herein incorporated by reference in its entirety in accordance with 37c.f.r. § 1.52(e) (5).

Background

Drug inhibitors targeting components of the mitogen-activated protein kinase (MAPK) signaling pathway show clinical efficacy in a variety of cancers, particularly those with mutations in the BRAF protein kinase. Both RAF and MEK inhibitors are approved as single agents for use in late metastatic BRAF mutant melanoma. BRAF and MEK inhibitor activity, alone or in combination, is unpredictable in other cancers, but has promising efficacy in BRAF mutant thyroid and lung cancers, with only mild activity in BRAF mutant colorectal cancers.

As with other targeted therapies, the pattern of disease response to RAF and MEK inhibitors appears to be influenced by the inherent genetic heterogeneity present in cancer using drugs. For example, it has been shown that certain genetic alterations, including PTEN and other changes that activate the PI3K cell growth signaling pathway, can predict poor initial response and/or relatively rapid progression in BRAF mutant melanoma treated with the RAF inhibitor vemurafenib. Also, direct mutations in the locus of the MEK gene appear to occur in tumors that progress after BRAF, MEK or combination drug therapy. Several other examples from RAS and RAF gene amplification and splicing mutations indicate that acquired drug resistance occurs when oncogenic pleiotropic effects are encountered under selective pressure for targeted drug therapy.

In view of the foregoing, there is a need for new targeted agents that would ideally inhibit multiple nodes of oncogenic pathways, and also be effective in combination by inducing a selective pressure burden that exceeds the adaptability of multiple cancer genomes. The present application is directed to meeting these and other needs.

Summary of The Invention

One embodiment of the invention is a method for treating or ameliorating the effects of a cancer that is refractory or resistant to non-ERK MAPK pathway inhibitor therapy in a subject. The method comprises administering to the subject an effective amount of BVD-523 or a pharmaceutically acceptable salt thereof.

Another embodiment of the invention is a method of treating or ameliorating the effects of cancer in a subject. The method comprises the following steps:

(a) identifying a patient having a cancer that becomes refractory or resistant to a BRAF inhibitor therapy, a MEK inhibitor therapy, or both BRAF and MEK inhibitor therapy; and

(b) administering to a subject having the refractory or resistant cancer an effective amount of an ERK inhibitor that is BVD-523 or a pharmaceutically acceptable salt thereof.

Another embodiment of the invention is a method of treating or ameliorating the effects of a cancer that is refractory or resistant to BRAF inhibitor therapy, MEK inhibitor therapy, or both in a subject. The method comprises administering to the subject an effective amount of BVD-523 or a pharmaceutically acceptable salt thereof.

Another embodiment of the invention is a method of identifying a subject with cancer who will benefit from therapy with an ERK inhibitor. The method comprises the following steps:

(a) obtaining a biological sample from a subject; and

(b) screening the sample to determine whether the subject has one or more of the following markers:

(i) the transition between the RAF isoforms is carried out,

(ii) upregulation of Receptor Tyrosine Kinase (RTK) or NRAS signaling,

(iii) reactivation of mitogen-activated protein kinase (MAPK) signaling,

(iv) the presence of a mutation that activates the activation of MEK,

(v) amplification of the mutant BRAF is carried out,

(vi) the up-regulation of STAT3 is,

(vii) mutations in the allosteric pocket of MEK that either directly block binding of the inhibitor to MEK or result in constitutive MEK activity,

wherein the presence of one or more of the markers confirms that the cancer in the subject is refractory or resistant to treatment with a BRAF and/or MEK inhibitor and that the subject would benefit from treatment with an ERK inhibitor that is BVD-523 or a pharmaceutically acceptable salt thereof.

Another embodiment of the invention is a pharmaceutical composition for treating or ameliorating the effects of a cancer in a subject that is refractory or resistant to non-ERK MAPK pathway therapy. The composition comprises a pharmaceutically acceptable carrier or diluent and an effective amount of BVD-523 or a pharmaceutically acceptable salt thereof.

Another embodiment of the invention is a kit for treating or ameliorating the effects of a cancer that is refractory or resistant to non-ERK MAPK pathway therapy in a subject. The kit comprises any pharmaceutical composition according to the invention, packaged with instructions for its use.

Another embodiment of the invention is a method for inhibiting phosphorylation of RSK in a cancer cell that is refractory or resistant to a non-ERK MAPK pathway inhibitor. The method comprises contacting the cancer cells with an effective amount of BVD-523 or a pharmaceutically acceptable salt thereof for a time sufficient to inhibit RSK phosphorylation in the cancer cells.

Another embodiment of the invention is a method of treating a subject with unresectable or metastatic BRAF600 mutation positive melanoma comprising administering to the subject 600mg BID of BVD-523 or a pharmaceutically acceptable salt thereof.

Another embodiment of the invention is a composition comprising 600mg BVD-523 or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier, adjuvant or vehicle for use in the treatment of a subject with unresectable or metastatic BRAF600 mutation positive melanoma.

Brief Description of Drawings

This patent or application document contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the office upon request and payment of the necessary fee.

Fig. 1A-1C show the progression of dose escalation studies in human malignant melanoma cell lines (a375 cells) at month 1. Various treatments (trametinib (MEK inhibitor type 2), dabrafenib (BRAF inhibitor) and BVD-523(ERK1/2 inhibitor)) are shown as markers.

Fig. 2A-2H show the results of proliferation assays that track changes in sensitivity to increasing agents at month 1. The various treatments (trametinib, dabrafenib, BVD-523, and paclitaxel) are shown as markers at the top of the graph. The headings on the right side of the figure show the various types of cells generated by the dose escalation study. For example, "dabrafenib" refers to cells that have been treated with the highest dose of dabrafenib from month 1 of the dose escalation study. Parental refers to control cells that have not been treated with drug. Fig. 2A, 2C, and 2G were normalized to the control, while fig. 2D, 2F, and 2H show the raw data.

Fig. 3A-3D show the progression of dose escalation studies in a375 cells at month 2. The various treatments (trametinib, dabrafenib and BVD-523) are indicated by the markers.

Fig. 4A-4H show the results of proliferation assays that track changes in sensitivity to increasing agents at month 2. The various treatments (trametinib, dabrafenib, BVD-523, and paclitaxel) are shown as markers at the top of the graph. The headings on the right side of the figure show the various types of cells generated by the dose escalation study. For example, "dabrafenib" refers to cells that have been treated with the highest dose of dabrafenib since month 2 of the dose escalation study. Parental refers to control cells that have not been treated with drug. Fig. 4A, 4C, and 4G were normalized to the control, while fig. 4D, 4F, and 4H show the raw data.

Fig. 5A-5H show only parental and BVD-523 cell line data from fig. 4A-4H. Various treatments (trametinib, dabrafenib, BVD-523, and paclitaxel) are indicated as markers. Fig. 5A, 5C, and 5G were normalized to the control, while fig. 5D, 5F, and 5H show the raw data.

Fig. 6A-6D show the progression of dose escalation studies in human malignant cell lines at month 3 (a375 cells). The various treatments (trametinib, dabrafenib and BVD-523) are indicated by the markers.

Figure 7 is a histogram showing the proliferation assay results applied to cells grown in DMSO control wells from the dose escalation assay.

Fig. 8A-8D are a set of line graphs showing proliferation assays at month 3 of the study. The various treatments (trametinib, dabrafenib, BVD-523, and paclitaxel) are shown as markers at the top of the graph. The headings on the right side of the figure show the various types of cells generated by the dose escalation study. For example, "dabrafenib" refers to cells treated with the highest dose of dabrafenib starting at month 3 of the dose escalation study. Parental refers to control cells that have not been treated with drug.

Fig. 9A-9D show only parental, dabrafenib and BVD-523 cell line data from fig. 8A-8D.

Figure 10A is a dose matrix showing% inhibition of trametinib/dabrafenib combination in a375 cells using Alamar Blue cell viability assay. Figure 10B is a dose matrix showing trametinib/dabrafenib combinations over Bliss overdose. Fig. 10C and 10D show% viability of dalafenib and trametinib single agent treatment in a375 cells using Alamar Blue cell viability assay versus control treated with DMSO alone. Figure 10E shows the% viability of the combination of dabrafenib and trametinib in a375 cells using the Alamar Blue cell viability assay versus the control treated with DMSO alone.

FIG. 11A is a dose matrix showing% inhibition of trametinib/dabrafenib combination in A375 cells using the CellTiter-Glo cell viability assay. Figure 11B is a dose matrix showing that the trametinib/dabrafenib combination exceeded the Bliss excess. Fig. 11C and 11D show% viability of dabrafenib and trametinib single agent treatment in a375 cells using the CellTiter-Glo cell viability assay versus a control treated with DMSO alone. FIG. 11E shows the% viability of the combination of dabrafenib and trametinib in A375 cells using the CellTiter-Glo cell viability assay versus the control treated with DMSO only.

FIG. 12A is a dose matrix showing% inhibition of BVD-523/dabrafenib combination in A375 cells using Alamar Blue cell viability assay. FIG. 12B is a dose matrix showing BVD-523/dabrafenib combination in excess of Bliss. FIGS. 12C and 12D show% viability of Dalafinib and BVD-523 single agent treatment in A375 cells relative to a control treated with DMSO only using the Alamar Blue cell viability assay. FIG. 12E shows the% viability in A375 cells of the combination of dabrafenib and BVD-523 using Alamar Blue cell viability assay versus control treated with DMSO only.

FIG. 13A is a dose matrix showing% inhibition of BVD-523/dabrafenib combination in A375 cells using the CellTiter-Glo cell viability assay. FIG. 13B is a dose matrix showing BVD-523/dabrafenib combination in excess of Bliss. FIGS. 13C and 13D show% viability of Dalafinib and BVD-523 single agent treatment in A375 cells relative to a control treated with DMSO only using the CellTiter-Glo cell viability assay. FIG. 13E shows the% viability of the combination of dabrafenib and BVD-523 in A375 cells using the CellTiter-Glo cell viability assay versus the DMSO only control.

FIG. 14A is a dose matrix showing% inhibition of trametinib/BVD-523 combination in A375 cells using Alamar Blue cell viability assay. FIG. 14B is a dose matrix showing trametinib/BVD-523 in combination over Bliss overdose. FIGS. 14C and 14D show% viability of BVD-523 and trametinib single agent treatment in A375 cells relative to a control treated with DMSO only using the Alamar Blue cell viability assay. FIG. 14E shows the% viability of BVD-523 and trametinib combination in A375 cells using Alamar Blue cell viability assay versus control treated with DMSO only.

FIG. 15A is a dose matrix showing% inhibition of the combination trametinib/BVD-523 in A375 cells using the CellTiter-Glo cell viability assay. FIG. 15B is a dose matrix showing trametinib/BVD-523 combination over Bliss overdose. FIGS. 15C and 15D show% survival in A375 cells using the CellTiter-Glo cell viability assay versus control treated with DMSO only, BVD-523 and trametinib single agent treatment. FIG. 15E shows the% viability of BVD-523 and trametinib combination in A375 cells using the CellTiter-Glo cell viability assay versus the control treated with DMSO only.

Fig. 16A-16D are a set of images showing Western blot analysis of MAPK signaling in a375 cells after 4 hours of treatment with BVD523, dabrafenib (Dab) and Tram (Tram) at various concentrations (in nM). Unless otherwise stated, each lane was loaded with 40 μ g total protein. In this experiment, duplicate samples were collected. Fig. 16A and 16B show the results of duplicate samples. Similarly, fig. 16C and 16D also show the results for duplicate samples. In fig. 16A and 16B, pRSK1 has a relatively weak signal in a375 cells compared to other markers. Different pRSK1-S380 antibodies from CellSignaling (catalog No. 11989) were tested, but no detectable signal was given (data not shown). In FIGS. 16C and 16D, pCRAF-338 gives the minimum signal.

FIGS. 17A-17D are a set of images showing Western blot analysis of MAPK signaling in human colorectal cancer cell lines (HCT116 cells) after 4 hours of treatment with various concentrations (in nM) of BVD-523, dabrafenib (Dab) and trametinib (Tram). Unless otherwise stated, each lane was loaded with 40 μ g total protein. In this experiment, duplicate samples were collected. Fig. 17A and 17B show the results of duplicate samples. Similarly, fig. 17C and 17D also show the results for duplicate samples. In FIGS. 17A and 17B, pRSK1 levels appeared to be very low in HCT116 cells, and pCRAF-338 signals were also weak in FIGS. 17C and 17D.

Fig. 18A-18D are a set of images showing Western blot analysis of cell cycle and apoptosis signaling in a375 melanoma cells after 24 hours of treatment with BVD-523 ("BVD 523"), trametinib ("tram"), and/or dabrafenib ("Dab") at various concentrations (in nM) as markers. Each lane was loaded with 50 μ g total protein unless otherwise stated. In this experiment, duplicate samples were collected. Fig. 18A and 18B show the results of duplicate samples. Similarly, fig. 18C and 18D also show the results for duplicate samples. In FIGS. 18A and 18B, there is no band corresponding significantly to the size of cleaved PARP (89 kDa).

Fig. 19 shows that BVD-523 can treat acquired resistance to targeted drugs in vivo. Patient-derived line ST052C was isolated from BRAFV600E melanoma patients progressing after 10 months of treatment with MAPK pathway-directed therapy. Ex vivo treatment, ST052C showed acquired cross-resistance to 50mg/kg BID of dabrafenib. Meanwhile, BVD-523 was effective as a single agent at 100mg/kg BID in ST 052C.

Fig. 20 is a flow chart showing a dose escalation protocol used herein.

Figure 21 shows a schematic of the mitogen-activated protein kinase (MAPK) pathway.

FIGS. 22A-22E show the results of a single agent proliferation assay. Proliferation results are shown for treatment with BVD-523 (FIG. 22A), SCH772984 (FIG. 22B), dabrafenib (FIG. 22C), trametinib (FIG. 22D) and paclitaxel (FIG. 22E).

FIGS. 23A-23O show the results for the combination of BVD-523 and dabrafenib. FIG. 23A shows a dose matrix showing combined inhibition (%) in RKO parental cells. Fig. 23B-23C show the results of the combined single agent proliferation assay of fig. 23A. Fig. 23D shows the combined Loewe excess in fig. 23A, and fig. 23E shows the combined Bliss excess in fig. 23A. FIG. 23F shows a dose matrix showing combined inhibition (%) in RKO MEK1(Q56P/+) -clone 1 cells. Fig. 23G-fig. 23H show the results of the combined single agent proliferation assay in fig. 23F. Fig. 23I shows the Loewe excess combined in fig. 23F, and fig. 23J shows the Bliss excess combined in fig. 23F. FIG. 23K shows a dose matrix showing combined inhibition (%) in RKO MEK1(Q56P/+) -clone 2 cells. Fig. 23L-fig. 23M show the results of the combined single agent proliferation assay in fig. 23K. Fig. 23N shows the combined Loewe excess in fig. 23K, and fig. 23O shows the combined Bliss excess in fig. 23K.

Fig. 24A-24O show the results of the combination of SCH772984 and dabrafenib. FIG. 24A shows a dose matrix showing combined inhibition (%) in RKO parental cells. Fig. 24B-fig. 24C show the results of the combined single agent proliferation assay of fig. 24A. Fig. 24D shows the combined Loewe excess in fig. 24A, and fig. 24E shows the combined Bliss excess in fig. 24A. FIG. 24F shows a dose matrix showing combined inhibition (%) in RKO MEK1(Q56P/+) -clone 1 cells. Fig. 24G-fig. 24H show the results of the combined single agent proliferation assay in fig. 24F. Fig. 24I shows the Loewe excess combined in fig. 24F, and fig. 24J shows the Bliss excess combined in fig. 24F. FIG. 24K shows a dose matrix showing combined inhibition (%) in RKO MEK1(Q56P/+) -clone 2 cells. Fig. 24L-fig. 24M show the results of the combined single agent proliferation assay in fig. 24K. Fig. 24N shows the combined Loewe excess in fig. 24K, and fig. 24O shows the combined Bliss excess in fig. 24K.

Fig. 25A-25O show results for combinations of trametinib and dabrafenib. FIG. 25A shows a dose matrix showing combined inhibition (%) in RKO parental cells. Fig. 25B-fig. 25C show the results of the combined single agent proliferation assay of fig. 25A. Fig. 25D shows the combined Loewe excess in fig. 25A, and fig. 25E shows the combined Bliss excess in fig. 25A. FIG. 25F shows a dose matrix showing combined inhibition (%) in RKO MEK1(Q56P/+) -clone 1 cells. Fig. 25G-fig. 25H show the results of the combined single agent proliferation assay in fig. 25F. Fig. 25I shows the Loewe excess combined in fig. 25F, and fig. 25J shows the Bliss excess combined in fig. 25F. FIG. 25K shows a dose matrix showing combined inhibition (%) in RKO MEK1(Q56P/+) -clone 2 cells. Fig. 25L-fig. 25M show the results of the combined single agent proliferation assay in fig. 25K. Fig. 25N shows the combined Loewe excess in fig. 25K, and fig. 25O shows the combined Bliss excess in fig. 25K.

Fig. 26A shows the combined Lowe volumes tested. Figure 26B shows the combined Bliss volumes tested. Figure 26C shows the synergy score for the tested combinations.

Figures 27A-27I show changes in MAPK and effector pathway signaling in MEK acquired resistance. Syngeneic RKO parent and MEK1(Q56P/+) cells were treated with compounds for 4 or 24 hours and then immunoblotted with the indicated antibodies. Dabrafenib is a BRAF inhibitor and trametinib is a MEK inhibitor. FIG. 27A shows increased signaling in RKO MEK1(Q56P/+) cells. FIGS. 27B-27C show the results of 4 hours treatment in experiment 1 (see example 7) in RKO parental (27B) and RKO MEK1(Q56P/+) (27C) cells. FIGS. 27D-27E show the results of 4 hour treatment in experiment 2 (see example 7) in RKO parental (27D) and RKO MEK1(Q56P/+) (27E) cells. FIGS. 27F-27G show the results of 4 hour treatment in experiment 2 (see example 7) in RKO parental (27F) and RKO MEK1(Q56P/+) (27G) cells. FIGS. 27H-27I show a summary of the results in RKO parent (27H) and RKO MEK1(Q56P/+) (27I) cells.

FIGS. 28A-28E show the results of the combination of BVD-523 and SCH 772984. Fig. 28A shows a dose matrix showing combined inhibition (%) in a375 cells. Fig. 28B-28C show the results of the combined single agent proliferation assay of fig. 28A. Fig. 28D shows the Loewe excess combined in fig. 28A. FIG. 28E shows the combined Bliss excess in FIG. 28A.

FIGS. 29A-29F show the discovery and characterization of the novel ERK1/2 inhibitor BVD-523 (ulixentinib). FIG. 29A shows that BVD-523 exhibits inhibition in a reversible ATP competition. This is achieved by inhibiting the IC of ERK2 as the ATP concentration increases₅₀A linear increase in value is demonstrated as shown in fig. 29B. FIG. 29C shows a representative plot (plot) of the dose-response curve, and FIG. 29D shows the IC₅₀Plot over time. FIG. 29E shows the binding of BVD-523 to ERK2 and phospho-ERK 2(pERK2) compared to the negative control protein p 38. FIG. 29F shows the binding of BVD-523 to ERK2 compared to the ERK inhibitors SCH772984 and pyrazolyl pyrroles.

Fig. 30A-fig. 30D show that BVD523 inhibits cell proliferation and enhances caspase 3 and caspase 7 activity in vitro. Fig. 30A shows that BVD-523 shows preferential activity in cells with MAPK pathway mutations, as defined by the presence of mutations in RAS family members and RAF. In addition, as shown in FIG. 30B, BVD-523 blocked sensitive cell line in G1 phase of cell cycle. FIG. 30C shows that BVD-523 induces A375, WM266 and LS411 after 72 hours of exposureThe concentration and time dependence of caspase activity increased in the N cancer cell lines. FIG. 30D shows MAPK pathway and effector protein trafficking at BRAF^V600EAcute (4 hours) and prolonged (24 hours) BVD-523 treatment in mutant A375 cells.

FIGS. 31A-31C show the antitumor activity of BVD-523 in vivo. BVD-523 monotherapy inhibited tumor growth in A375 (FIG. 31A) and Colo205 cell line xenograft models (FIG. 31B) ((R))^aP<0.0001, compared to vehicle control; CPT-11 was administered only on days 14 and 18). Abbreviations: BID, twice daily; CMC, carboxymethyl cellulose; QD, daily; Q4D, once every 4 days. FIG. 31C shows that increased phosphorylation of ERK1/2 correlates with BVD-523 concentration in Colo205 xenografts.

FIG. 32A shows the signaling effect of ERK1/2 inhibitors. Using RPPA, the effect on proteins was measured in cell lines (A375, AN3Ca, Colo205, HCT116, HT29 and MIAPaca2) after treatment with the ERK1/2 inhibitor BVD-523(BVD), Vx11e (Vx), GDC-0994(GDC) or SCH722984 (SCH). FIG. 32B shows that the ERK inhibitors BVD-523, GDC-0994 and Vx11e have different effects on phospho-ERK (ERK 1/2T 202Y 204) compared to SCH 722984; phospho-RSK (p90 RSK 380) and cyclin D1 were tested for inhibition by ERK inhibitors. Abbreviations: BRAFi, BRAF inhibitors; MEKi, MEK inhibitor. FIG. 32C shows a western blot assay of cell and nuclear fractions from RKO cell line after treatment with BVD-523, trametinib, SCH722984, or dabrafenib. Histone H3 (nuclear localization protein) and HSP90 (cytoplasmic localization protein) were included as positive controls to confirm that the nuclear and cytoplasmic fractions were properly enriched; the nuclear fraction has high H3 and the cytoplasmic fraction has higher HSP 90.

FIG. 33 shows that ERK inhibitors BVD-523, Vx11, GDC-0994 and SCH772984(SCH) show cell-line dependent changes in phospho-ATK levels. Abbreviations: DMSO, dimethylsulfoxide.

FIGS. 34A-34D show that BVD-523 shows activity in a resistance model of BRAF/MEK inhibition. Shows BRAF following exposure to increasing concentrations of drug^V600EApparent resistance to BVD-523, dabrafenib or trametinib in A375 cells. A set of strict "criteria" was applied to determine when the dose could be increased to ensure that the kinetics of resistance between treatments were comparable. See example 1. For IC₅₀The multiple of (d) indicates time; each point on the plotted line represents a change in culture medium or cell distribution. FIG. 34A shows that acclimating cells to growth in the presence of BVD-523 is more challenging than using dabrafenib or trametinib. Fig. 34B shows that BVD-523 sensitivity was retained in a375 cells cultured to obtain resistance to combined BRAF (dabrafenib) + MEK (trametinib) inhibition. In FIG. 34C, cells were treated with compounds for 96 hours and CellTiter-Viability was assessed. Due to MEK1^Q56PIn BRAF (dabrafenib) and MEK (trametinib) inhibitors^V600EBVD-523 activity was retained in RKO cells. FIG. 34D shows BRAK^V600EBVD-523 inhibition of pRSK in the mutant cell line RKO in MEK1^Q56PMaintained in the presence of MEK1^Q56PConferring resistance to inhibition by MEK and BRAF. Knock-in of KRAS mutant allele into SW48 cell line significantly reduced sensitivity to the MEK inhibitors trametinib and semetinib (selumetinib), while retaining relative sensitivity to BVD-523.

Fig. 35A shows BVD-523 in vivo activity in xenografts from vemurafenib (vemurafenib) relapsed patients. The mean tumor volumes (. + -. SEM) of BVD-523100 mg/kg BID alone, dabrafenib 50mg/kg BID alone, and BVD-523100 mg/kg BID plus dabrafenib 50mg/kg BID are shown. Abbreviations: BID, twice daily; SEM, standard error of mean.

Fig. 36A-36D show the benefit of combined BVD-523 and BRAF inhibition. FIGS. 36A-36B show the initial volume at tumor of 75-144mm³A375BRAF of^V600EBVD-523 plus dabrafenib in a xenograft model of a melanoma cell line compared to treatment with either agent aloneThe combination shows excellent antitumor activity. FIGS. 36C-36D show the results from the larger tumor volume at the start of dosing (700-800 mm)³) Similar data of the same model. Plots of mean tumor growth (left panel) and Kaplan-Meier survival (right panel) are presented for each study. Abbreviations: BID, twice daily; QD, once daily.

Figure 37A shows that in SW48 colorectal cells engineered with KRAS alleles, the response to paclitaxel was unchanged compared to controls. FIG. 37B shows the combined interaction between BVD-523 and Vemurafenib, evaluated using the LoeweAdditity and Bliss Independence models using an 8 × 10 concentration matrix and analyzed using the Horizon's Chalice, Bioinformatics Software. By showing that calculated excess inhibition exceeds the expectation of addition in the dose matrix as a heat map, and by reporting a quantitative "synergy score" based on the Loewe model, Chalice can identify potential synergistic interactions. The results indicate that the interaction between BVD-523 and vemurafenib is at least additive and in some cases carries BRAF^V600ESynergistic in the mutant melanoma cell line. FIG. 37C shows that BVD-523 in combination with dabrafenib significantly delayed BRAF at A375^V600EOnset of acquired resistance in melanoma cells. Temporal acquisition of resistance in response to increasing concentrations of dabrafenib alone or in combination with BVD-523 or trametinib was evaluated. Strict criteria apply as to when the dose can be increased to ensure that the adaptation kinetics are comparable between treatments. See example 1.

FIG. 38 shows that BVD-523 inhibits ex vivo PMA-stimulated phosphorylation of RSK1/2 in human whole blood. The average of the BVD-523 concentration data set is indicated by (-). For each concentration of BVD-523, n is 20. Abbreviations: PBMC, peripheral blood mononuclear cells; RSK, ribosomal S6 kinase.

FIG. 39A shows steady-state BVD-523 pharmacokinetics (cycle 1, day 15). Red dotted line represents EC₅₀200ng/mL HWB. Abbreviations: AUC, area under the curve; BID, twice daily; c_maxMaximum concentration; EC (EC)₅₀50% maximum effectiveConcentration; HWB, human whole blood; SD, standard deviation. FIG. 39B shows the pharmacodynamic inhibition of ERK phosphorylation by BVD-523 in human whole blood. Abbreviations: BID, twice daily; pRSK, phospho-RSK; RSK, ribosomal S6 kinase.

Fig. 40A shows the optimal radiographic response in patients treated with BVD-523. All patients with disease as measured by recistv1.1 who received a dose ≧ 1 dose of study treatment and had >1 in-treatment tumor assessment (25/27; 2 scans not receiving two target lesions). Responses were measured as the sum of the longest diameters of each target lesion from baseline. The dose displayed is the dose that the patient received in response. The dashed line represents the threshold for the partial response according to RECIST v 1.1. Abbreviations: CRC, colorectal cancer; NET, neuroendocrine tumor; NSCLC, non-small cell lung cancer; NSGCT, non-seminal germ cell tumor; PNET, pancreatic NET; PTC, papillary thyroid carcinoma; RECIST v1.1, response assessment criteria for solid tumors version 1.1; SLD, sum of maximum diameters. Fig. 40B shows a computed tomography scan of confirmed partial response in 61 year old patients with BRAF mutant melanoma treated with BVD-523.

Figure 41 shows tumor response and tumor progression. A swimmer plot (swimmer plot) showing that response to treatment with BVD-523 can assess tumor response, tumor progression, and treatment duration in patients is shown. The origin of the vertical axis corresponds to the randomization date or the reference start date. Analysis expiration date: 12 months and 1 day 2015. Abbreviations: BID, twice daily.

Detailed Description

As used herein, the terms "treat," "treatment," "treat," and grammatical variations thereof refer to subjecting an individual subject to a protocol, regimen, process, or remedy in which a physiological response or result is desired in the subject (e.g., patient). In particular, the methods and compositions of the invention can be used to slow the development of disease symptoms or delay the onset of a disease or condition, or stop the progression of disease development. However, because each subject being treated may not respond to a particular treatment protocol, regimen, procedure, or remedy, treatment does not require that a desired physiological response or result be achieved in each subject or population of subjects (e.g., patient population). Thus, a given subject or population of subjects (e.g., a population of patients) may fail to respond or respond insufficiently to treatment.

As used herein, the terms "ameliorate," "alleviating," and grammatical variants thereof refer to reducing the severity of a disease symptom in a subject.

As used herein, a "subject" is a mammal, preferably a human. In addition to humans, mammalian species within the scope of the present invention include, for example, farm animals, domestic animals, laboratory animals, and the like. Some examples of farm animals include cattle, pigs, horses, goats, and the like. Some examples of domestic animals include dogs, cats, and the like. Some examples of laboratory animals include primates, rats, mice, rabbits, guinea pigs, and the like.

In the present invention, BVD-523 corresponds to a compound according to formula (I):

BVD-523 can be synthesized according to published methods, for example, U.S. Pat. No. 7,354,939. The enantiomer of BVD-523 and racemic mixtures of the two enantiomers are also included in the scope of the present invention. BVD-523 is an ERK1/2 inhibitor whose mechanism of action is believed to be, for example, unique and distinct from certain other ERK1/2 inhibitors (e.g., SCH772984) and the pyrimidine structures used by hatzivasssiliou et al (2012). For example, other ERK1/2 inhibitors (e.g., SCH772984) inhibit the autophosphorylation of ERK (Morris et al, 2013), while BVD-523 allows autophosphorylation of ERK while still inhibiting ERK. (see, e.g., FIG. 18).

As used herein, the words "resistant" and "refractory" are used interchangeably. "resistant" to non-ERK MAPK pathway inhibitor therapy refers to a decrease in the efficacy of a non-ERK MAPK inhibitor in treating cancer.

As used herein, a "non-ERK MAPK inhibitor" refers to any substance that decreases the activity, expression or phosphorylation of a protein or other member of the MAPK pathway (which results in a decrease in cell growth or an increase in cell death), except for an ERK1/2 inhibitor. As used herein, "ERK 1/2 inhibitors" refers to those substances that (i) interact directly with ERK1 and/or ERK2, e.g., by binding to ERK1/2 and (ii) reduce the expression or activity of ERK1 and/or ERK2 protein kinases. Thus, inhibitors acting upstream of ERK1/2, such as MEK inhibitors and RAF inhibitors, are not ERK1/2 inhibitors according to the invention (but they are non-ERK MAPK inhibitors). Non-limiting examples of ERK1/2 inhibitors according to the invention include AEZS-131(Aeterna Zentaris), AEZS-136(Aeterna Zentaris), BVD-523(BioMed Valley Discovieres, Inc.), SCH-722984(Merck & Co.), SCH-772984(Merck & Co.), SCH-900353(MK-8353) (Merck & Co.), pharmaceutically acceptable salts thereof, and combinations thereof.

Briefly, with respect to the ERK1/2 module (light purple box) in fig. 21, the MAPK 1/2 signaling cascade is activated by ligand binding to Receptor Tyrosine Kinases (RTKs), briefly, with respect to the ERK1/2 module in fig. 21 (light purple box), the activated receptor recruits and phosphorylates the adaptor proteins Grb2 and SOS, which then interact with and cause activation of membrane-bound gtpase Ras, in its activated GTP-bound form, Ras recruits and activates RAF kinases (a-RAF, B-RAF and C-RAF/RAF-1) the activated RAF kinase activates MAPK 1/2(MKK 874 1/2), which in turn catalyzes the phosphorylation of threonine and tyrosine residues in the activation sequence Thr-Glu-Tyr of ERK1/2, with respect to the JNK/p kinase module (MAP 338 in fig. 21), upstream of MAPK 38, e.g. MEKK 364, mtk 38, and tyrosine residues in MAP 38, MAP 38-38, MAP 38, and MAP 38, including MAP 38.

Non-limiting examples of non-ERK MAPK pathway inhibitors according to the present invention include RAS inhibitors, RAF inhibitors (e.g., inhibitors of a-RAF, B-RAF, C-RAF (RAF-1)), MEK inhibitors, and combinations thereof. Preferably, the non-ERK MAPK pathway inhibitor is a BRAF inhibitor, a MEK inhibitor, and combinations thereof.

As used herein, "RAS inhibitors" refers to those substances that (i) interact directly with RAS, e.g., by binding to RAS and (ii) decrease RAS expression or activity. Non-limiting exemplary RAS inhibitors include, but are not limited to, farnesyl transferase inhibitors (e.g., tipifarnib and lonafarnib), farnesyl-containing small molecules (e.g., salirasib and TLN-4601), DCAI, as disclosed by Maurer (Maurer et al, 2012), Kobe0065 and Kobe2602, as disclosed by Shima (Shima et al, 2013), HBS 3(Patgiri et al, 2011), and AIK-4 (alllinky).

As used herein, "RAF inhibitors" refer to those substances that (i) interact directly with RAF, e.g., by binding RAF and (ii) decrease the expression or activity of RAF, e.g., a-RAF, B-RAF and C-RAF (RAF-1). Non-limiting exemplary RAF inhibitors, including BRAF inhibitors, include:

compound 7(Li et al),

compound 9(in the same way as above),

compound 10(in the same way as above),

compound 13(in the same way as above),

compound 14(in the same way as above),

compound 15(in the same way as above),

compound 16(in the same way as above),

compound 18(in the same way as above),

compound 19(in the same way as above),

compound 20(in the same way as above),

compound 21(in the same way as above),

compound 22(in the same way as above),

compound 23(in the same way as above),

compound 24(in the same way as above),

compound 25(in the same way as above),

compound 26(in the same way as above),

compound 27(in the same way as above),

compound 28(in the same way as above),

compound 30(in the same way as above),

compound 31(in the same way as above),

compound 32(in the same way as above),

compound 33(in the same way as above),

compound 34(in the same way as above),

compound 35(in the same way as above),

compound 36(in the same way as above),

compound 37(in the same way as above),

compound 38

(in the same way as above),

compound 39(in the same way as above),

compound 40(in the same way as above),

AAL881 (noval); AB-024(Ambit Biosciences), ARQ-736(ArQule), ARQ-761(ArQule), AZ628(Axon Meschem BV), BeiGene-283(BeiGene), BIIB-024(MLN 2480) (Sunesis & Takeda), b-RAF inhibitor (Sareum), BRAF kinase inhibitor (Selexagen Therapeutics), BRAF siRNA 313(tacaccagcaagctagatgca) and 523(cctatcgttagagtcttcctg) (Liu et al, 2007), CTT239065 (cancers institute), Dalafenib (GSK2118436), DP-4978 (Decipherara Pharmaceutica), HM-95573(Hanmi), GDC-0879(Genentech), GW-5074(Sigma Aldrich), ISIS 5132(Novartis), L947750 (Merck), Pharma (LERTI), LERARTN (AOXbax 5568), Pluronic (Plurex-32068), Plurex-32068 (Plurex-32068), Plurex-47265 (Plurex), RO5126766 (Hoffmann-LaRoche), SB 590885(GlaxoSmithKline), SB699393(GlaxoSmithKline), sorafenib (OnyxPharmaceutics), TAK 632(Takeda), TL-241(Teligene), Verofibrib (RG7204 or PLX4032) (Daiichi Sankyo), XL-281(Exelixis), ZM-336372(AstraZeneca), pharmaceutically acceptable salts thereof, and combinations thereof.

As used herein, "MEK inhibitor" refers to those substances that (i) interact directly with MEK, e.g., by binding to MEK, and (ii) reduce the expression or activity of MEK. Thus, inhibitors acting upstream of MEK, such as RAS inhibitors and RAF inhibitors, are not MEF inhibitors according to the present invention. Non-limiting examples of MEK inhibitors include anthrax toxin, antroquinol (antroquinonol) (Golden Biotechnology), ARRY-142886(6- (4-bromo-2-chloro-phenylamino) -7-fluoro-3-methyl-3H-benzimidazole-5-carboxylic acid (2-hydroxy-ethoxy) -amide) (Array BioPharma), ARRY-438162(Array BioPharma), AS-1940477(Astellas), AS-703988(Merck KGaA), bentamaimod (Merck KGaA), BI-847325 (Boehrer Ingelheim), E-6201(Eisai), GDC-0623(Hoffmann-La Roche), GDC-0973 (cobimetinib) (Hoffmann-La), L783277(Merck), the lethal factor of the Alxon (Biomethyloxynaphthalene-094-MEK) and MEK-094' -lethal factor (Biomethyloxynaphthalene-PD) (Biomethyloxy-PD-094-PD), and MEK-3559 (Biomethyloxy-L-H-L-5-A) -ketone) (Pfizer), PD 184352(CI-1040) (Pfizer), PD-0325901(Pfizer), pimasetib (Santera pharmaceuticals), RDEA119(Ardea biosciences/Bayer), refametinib (AstraZeneca), RG422(Chugai Pharmaceutical Co.), RO092210(Roche), RO4987655(Hoffmann-La Roche), RO 51766 (Hoffmann-La Roche), Semetinib (AZD6244) (AstraZeneca), SL327(Sigma), TAK-733(Takeda), Trimetinib (Japan Tobaco), U0126(1, 4-diamino-2, 3-dicyano-1, 4-bis (2-aminophenylthio) butadiene) (Sigma), WX-554 (Wilkeex), Yopex J (Mittopoco), polypeptides 2010, and pharmaceutically acceptable salts thereof.

In one aspect of this embodiment, substantially all phosphorylation of ribosomal s6 kinase (RSK) is inhibited following administration of BVD-523 or a pharmaceutically acceptable salt thereof. As used herein in the context of RSK phosphorylation, "substantially all" refers to a reduction of greater than 50%, preferably greater than 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% reduction.

In another aspect of this embodiment, the cancer has MAPK activity. As used herein, having "MAPK activity" means that the protein downstream of ERK is still active, even though the protein upstream of ERK may be inactive. Such cancer may be a solid tumor cancer or a hematologic cancer.

In the present invention, cancer includes solid and hematologic cancers. Non-limiting examples of solid cancers include adrenocortical carcinoma, anal carcinoma, bladder carcinoma, bone cancer (e.g., osteosarcoma), brain cancer, breast cancer, carcinoid, carcinoma (carcinoma), cervical cancer, colon cancer, endometrial cancer, esophageal cancer, extrahepatic bile duct cancer, Ewing family of cancers, extracranial germ cell cancer, eye cancer, gallbladder cancer, stomach cancer, germ cell tumor, gestational trophoblastic tumor, head and neck cancer, hypopharynx cancer, islet cell cancer, kidney cancer, large intestine cancer, larynx cancer, leukemia, lip and oral cancer, liver tumor/cancer, lung tumor/cancer, lymphoma, malignant mesothelioma, Merkel cell cancer, mycosis fungoides, myelodysplastic syndrome, myeloproliferative disorders, nasopharyngeal carcinoma, neuroblastoma, oral cancer, oropharyngeal cancer, osteosarcoma, epithelial ovarian carcinoma, germ cell cancer, pancreatic cancer, sinus and nasal cavity cancer, parathyroid gland cancer, penile cancer, pituitary cancer, plasma cell tumors, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, transitional cell carcinoma of the renal pelvis and ureter, salivary gland cancer, Sezary syndrome, skin cancer (such as cutaneous t-cell lymphoma, Kaposi's sarcoma, mast cell tumor and melanoma), small intestine cancer, soft tissue sarcoma, stomach cancer, testicular cancer, thymoma, thyroid cancer, urinary tract cancer, uterine cancer, vaginal cancer, vulval cancer and nephroblastoma.

Examples of hematologic cancers include, but are not limited to, leukemias, such as adult/childhood acute lymphocytic leukemia, adult/childhood acute myelogenous leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia, lymphomas, such as aids-related lymphoma, cutaneous T-cell lymphoma, adult/childhood hodgkin ' S lymphoma, mycosis fungoides, adult/childhood non-hodgkin ' S lymphoma, primary central nervous system lymphoma, S-zary syndrome, cutaneous T-cell lymphoma, and waldenstrom ' S macroglobulinemia, as well as other proliferative diseases, such as chronic myeloproliferative diseases, langerhans cell histiocytosis, multiple myeloma/plasma cell tumors, myelodysplastic syndrome, and myelodysplastic/myeloproliferative tumors.

Preferably, the cancer is selected from the group consisting of colorectal cancer, breast cancer, pancreatic cancer, skin cancer and endometrial cancer. More preferably, the cancer is melanoma.

In another aspect of this embodiment, the method further comprises administering to the subject at least one additional therapeutic agent effective to treat or ameliorate the effects of cancer. The additional therapeutic agent may be selected from the group consisting of an antibody or fragment thereof, a cytotoxic agent, a toxin, a radionuclide, an immunomodulator, a photoactive therapeutic agent, a radiosensitizer, a hormone, an anti-angiogenic agent, and a combination thereof.

As used herein, "antibody" includes naturally occurring immunoglobulins as well as non-naturally occurring immunoglobulins, including, for example, single chain antibodies, chimeric antibodies (e.g., humanized murine antibodies) and heteroconjugate antibodies (e.g., bispecific antibodies). Fragments of antibodies include those that bind antigen (e.g., Fab ', F (ab')₂Fab, Fv and rIgG). See also, e.g., Pierce Catalog and Handbook, 1994-; kuby, j., Immunology, 3rd ed., w.h.freeman&Co., New York (1998). The term antibody also includes bivalent or bispecific molecules, diabodies, triabodies and tetrabodies. The term "antibody" also includes polyclonal and monoclonal antibodies.

Examples of therapeutic antibodies useful in the present invention include rituximab (Rituxan), cetuximab (Erbitux), bevacizumab (Avastin), and Ibritumomab (Ibritumomab) (Zevalin).

Cytotoxic agents according to the present invention include DNA damaging agents, antimetabolites, antimicrotubule agents, antibiotic agents, and the like. DNA damaging agents include alkylating agents, platinum-based agents, intercalating agents, and DNA replication inhibitors. Non-limiting examples of DNA alkylating agents include cyclophosphamide, mechlorethamine, uramustine, melphalan, chlorambucil, ifosfamide, carmustine, lomustine, streptozotocin, busulfan, temozolomide, pharmaceutically acceptable salts, prodrugs and combinations thereof. Non-limiting examples of platinum-based agents include cisplatin, carboplatin, oxaliplatin, nedaplatin, satraplatin, triplatin tetranitrate, pharmaceutically acceptable salts, prodrugs, and combinations thereof. Non-limiting examples of intercalating agents include doxorubicin, daunorubicin, idarubicin, mitoxantrone, pharmaceutically acceptable salts, prodrugs thereof, and combinations thereof. Non-limiting examples of DNA replication inhibitors include irinotecan, topotecan, amsacrine, etoposide phosphate, teniposide, pharmaceutically acceptable salts, prodrugs, and combinations thereof. Antimetabolites include folate antagonists such as methotrexate and premethopterin, purine antagonists such as 6-mercaptopurine, dacarbazine, and fludarabine, and pyrimidine antagonists such as 5-fluorouracil, arabinosyl cytosine, capecitabine, gemcitabine, decitabine, pharmaceutically acceptable salts, prodrugs, and combinations thereof. Anti-microtubule agents include, but are not limited to, vinca alkaloids, paclitaxel (paclitaxel) Docetaxel (docetaxel: (b))) And ixabepilone (b)). Antibiotic agents include, but are not limited to, actinomycin, anthracycline, valrubicin, epirubicin, bleomycin, mithramycin, mitomycin, pharmaceutically acceptable salts, prodrugs and combinations thereof.

Non-limiting examples of inhibitors of the PI 3/Akt Pathway include A-674563(CAS # -73-2), AGL 2263, AMG-319 (Ampen, Thousandoando, Calca), AS- (5-benzo [1,3] dioxol-5-ylmethylene-thiazolidine-2, 4-dione), AS- (5- (2, 2-difluoro-benzo [1,3] dioxol-5-ylmethylene) -thiazolidine-2, 4-dione), AS-605240 (5-quinoxaline-6-methylene-1, 3-thiazolidine-2, 4-dione), AT7867 (CAS-00-1), benzimidazole series, genetech (Roche Holdings dibasic, Cas Sanwich Francisco, BML 257(CAS # 325), CAS # 120 (CAS #, CAS # WO 7, Californike et al, Californike # WO 1, Californike # WO 7, Californike-K), Californike-A-7, Californike, California-7, Californike, Californik, Californike, California-7, Californike, California-7, Californik, California-7, Californike, Californik, California-7, Californik, California-7, Californik, California-7, Californike, Californik, California-7, Californik, California-7, Californik, California-7, California-inhibitors, Californik, California-7, Californik, California-K, Californik, California-7, Californik, California-inhibitors, Californik, California-I, Californik, California-Californik, California-7, Californik, California-Californik, California-K, California-Californik, California-7, Californik, California-K, Californik, California-Californik, California.

In the present invention, the term "toxin" refers to an antigenic poison or venom of plant or animal origin. One example is diphtheria toxin or a portion thereof.

In the present invention, the term "radionuclide" refers to a radioactive substance that is administered to a patient, for example, intravenously or orally, after which it permeates into the target organ or tissue through the patient's normal metabolism, where it delivers local radiation for a short period of time. Examples of radionuclides include, but are not limited to, I-125, At-211, Lu-177, Cu-67, I-131, Sm-153, Re-186, P-32, Re-188, In-114m, and Y90.

In the present invention, the term "immunomodulator" refers to a substance that alters an immune response by enhancing or reducing the ability of the immune system to produce antibodies or primed cells that recognize and react with the antigen from which their production was initiated. The immunomodulator may be a recombinant, synthetic or natural agent, and includes cytokines, corticosteroids, cytotoxic agents, thymosins and immunoglobulins. Some immunomodulators occur naturally in the body, and some of them are useful in pharmacological formulations. Examples of immunomodulators include, but are not limited to, granulocyte colony-stimulating factor (G-CSF), interferon, imiquimod and cell membrane fractions from bacteria, IL-2, IL-7, IL-12, CCL3, CCL26, CXCL7, and synthetic cytosine phosphate guanosine (CpG).

In the present invention, the term "light-activated therapeutic agent" refers to compounds and compositions that become active upon exposure to light. Certain examples of light activated therapeutic agents are disclosed in, for example, U.S. patent application serial No. 2011/0152230 a1, "photoactive metal nitrosyl groups for blood pressure regulation and cancer therapy.

In the present invention, the term "radiosensitizer" refers to a compound that makes tumor cells more sensitive to radiotherapy. Examples of radiosensitizers include (misonidazole), metronidazole, tirapazamine (tirapazamine), and sodium trans-crocetinate.

In the present invention, the term "hormone" refers to a substance released by cells in one part of the body that affects cells in another part of the body. Examples of hormones include, but are not limited to, prostaglandins, leukotrienes, prostacyclin, thromboxanes, amylin, anti-mullerian hormone, adiponectin, adrenocorticotropic hormone, angiotensinogen, angiotensin, vasopressin, atrial natriuretic peptide (atriopeptin), brain natriuretic peptide, calcitonin, cholecystokinin, adrenocorticotropic hormone releasing hormone, enkephalin, endothelin, erythropoietin, follicle stimulating hormone, galanin, gastrin, ghrelin (ghrelin), glucagon, gonadotropin releasing hormone, growth hormone releasing hormone, human chorionic gonadotropin, human placental prolactin, growth hormone, inhibin, insulin, growth regulators, leptin, lipotropin (liptropin), luteinizing hormone, melanocyte stimulating hormone, motilin, orexin (orexin), oxytocin, pancreatic polypeptide, parathyroid hormone, prolactin, prolactin releasing hormone, relaxin, renin, secretin, somatostatin, thrombopoietin, thyroid stimulating hormone, testosterone, dehydroepiandrosterone, androstenedione, dihydrotestosterone, aldosterone, estradiol, estrone, estriol, cortisol, progesterone, calcitriol, and calcifediol.

Some compounds interfere with the activity of or prevent the production of certain hormones. These hormonal interfering compounds include, but are not limited to, tamoxifen (b), (c), (d) Anastrozole (a)) Letrozole (C)) And fulvestrant (). These compounds are also within the meaning of hormones in the present invention.

As used herein, an "anti-angiogenic" agent refers to a substance that reduces or inhibits the growth of new blood vessels, such as an inhibitor of Vascular Endothelial Growth Factor (VEGF) and an inhibitor of endothelial cell migration anti-angiogenic agents include, but are not limited to, 2-methoxyestradiol, angiostatin, bevacizumab, cartilage-derived angiostatic factor, endostatin, IFN- α -12, itraconazole, linamide (linomide), platelet factor-4, prolactin, SU5416, suramin, tasquinimod, tecogalan, tetrathiomolybdate, thalidomide, thrombospondin, TNP-470, ziv-aflibercept, pharmaceutically acceptable salts, prodrugs, and combinations thereof.

Another embodiment of the invention is a method for treating or ameliorating the effects of cancer in a subject. The method comprises the following steps:

(a) identifying a subject having a cancer that becomes refractory or resistant to a BRAF inhibitor therapy, a MEK inhibitor therapy, or both BRAF and MEK inhibitor therapy; and

Suitable and preferred subjects are as disclosed herein. In this embodiment, the method may be used to treat the cancer disclosed above. According to the invention, the cancer has MAPK activity.

In one aspect of this embodiment, identifying a subject having a cancer refractory or resistant to BRAF and/or MEK inhibitor therapy comprises:

(a) obtaining a biological sample from a subject; and

(b) screening the sample to determine whether the subject has become resistant to an inhibitor therapy selected from the group consisting of a BRAF inhibitor therapy, a MEK inhibitor therapy, and a combination thereof.

In the present invention, biological samples include, but are not limited to, blood, plasma, urine, skin, saliva, and biopsy samples. The biological sample is obtained from the subject by conventional procedures and methods known in the art.

Preferably, screening for cancers refractory or resistant to BRAF inhibitor therapy can include, for example, identifying (i) a switch between RAF isoforms, (ii) upregulation of RTK or NRAS signaling, (iii) reactivation of mitogen-activated protein kinase (MAPK) signaling, (iv) the presence of MEK activating mutations, and combinations thereof.

Switching between RAF isoforms can occur in subjects with acquired resistance to BRAF inhibitor therapy. To detect this switch, BRAF inhibitor resistant tumor cells can be recovered from the patient and assayed for ERK and phospho-ERK levels by Western blotting in the presence of BRAF inhibitors. Comparison with BRAF inhibitor-sensitive cells treated with BRAF inhibitors can reveal higher levels of phospho-ERK in BRAF inhibitor resistant tumor cells, which means that a switch has occurred in which another RAF isoform phosphorylates ERK but not BRAF. Confirmation of which RAF isoform has taken over may involve sh/siRNA mediated knockdown of ARAF and CRAF, respectively, in BRAF inhibitor resistant cells exposed to a BRAF inhibitor, followed by subsequent Western blotting for ERK and phospho-ERK levels. For example, if ara knockdown in BRAF inhibitor resistant cells exposed to a BRAF inhibitor still results in high levels of phospho-ERK, it indicates that CRAF has taken over to phosphorylate ERK. Likewise, if CRAF is knocked down in BRAF inhibitor resistant cells exposed to BRAF inhibitors and ERK is still highly phosphorylated, it means that ARAF has taken over ERK phosphorylation. RAF isotype switching may also involve the simultaneous knockdown of ARAF and CRAF in BRAF inhibitor resistant cells in the presence of BRAF inhibitors, effectively blocking all RAF-mediated phosphorylation. The resulting decrease in ERK phosphorylation indicates that BRAF inhibitor resistant cells have the ability to switch between RAF isoforms to phosphorylate ERK (villanuva et al, 2010).

The up-regulation of RTK or NRAS signaling may also be responsible for BRAF inhibitor resistance the detection may for example first involve the use of Western blot protocols with phosphate-specific antibodies to analyse the activation of the downstream RAF effector MEK1/2 and ERK1/2 if BRAF inhibitor resistant cells show high levels of activation of these proteins in the presence of BRAF inhibitors, it may be that RTK or NRAS up-regulation is responsible for it.

Detection of reactivation of MAPK signaling in BRAF inhibitor-resistant cells may indicate another alternative mechanism of BRAF inhibitor resistance. COT and C-RAF have been shown to be upregulated in the background of BRAF V600E exposed to BRAF inhibitors. Quantitative real-time RT-PCR can, for example, show increased COT expression in BRAF inhibitor resistant cells in the presence of BRAF inhibitors. Furthermore, sh/siRNA mediated knockdown of COT in BRAF inhibitor resistant cells in the presence of BRAF inhibitors may reduce the viability of BRAF inhibitor resistant cells, suggesting that these particular cells may be sensitive to COT inhibition and/or combination BRAF inhibitor/MEK inhibitor treatment (Johannessen et al, 2010).

Reactivation of MAPK signaling can also be accomplished in the context of BRAF inhibitor resistance by activating mutations in MEK 1. Massively parallel sequencing of the target of genomic DNA from BRAF inhibitor resistant tumors can reveal activating mutations in MEK1, such as C121S, G128D, N122D, and Y130, among others. Other silent mutations in MEK1 can be analyzed by expressing specific mutations, for example, in BRAF inhibitor sensitive cell lines such as a 375. Determining the level of growth inhibition in these cells upon exposure to a BRAF inhibitor can indicate whether the MEK1 mutation results in resistance to BRAF inhibitory therapy. To confirm this finding, Western blots of elevated levels of phospho-ERK 1/2 in cells ectopically expressing the MEK1 mutation may indicate that the MEK1 mutation allows BRAF inhibitor resistant tumors to bypass BRAF and promote phosphorylation of ERK by MEK1 (Wagle et al, 2011).

Screening for cancers that are refractory or resistant to MEK inhibitor therapy according to the present invention may include, for example, identifying (i) amplification of mutant BRAF, (ii) STAT3 upregulation, (iii) mutations in the allosteric pocket of MEK that directly block inhibitor binding to MEK or result in constitutive MEK activity, and combinations thereof.

Amplification of mutant BRAF can cause MEK inhibitor resistance. MEK inhibitor resistance is often associated with high levels of phosphorylated ERK and MEK in the presence of MEK inhibitors, which can be assessed, for example, by Western blotting. Amplification of mutant BRAF in MEK inhibitor resistant cell lines can be detected by, for example, Fluorescent In Situ Hybridization (FISH) or quantitative PCR of genomic DNA from resistant cell lines. Confirmation that BRAF expansion is the primary reason for MEK inhibitor resistance may require the use of BRAF-targeted sh/siRNA in resistant cells. If a significant decrease in MEK or ERK phosphorylation is observed, BRAF amplification may be a suitable target for further therapeutic approaches. (Corcoran et al, 2010).

Identification of STAT3 upregulation can indicate that a particular tumor sample is resistant to MEK inhibitor therapy. Genome-wide expression profiling may reveal that the STAT3 pathway is upregulated in tumors. Other techniques, such as Western blotting for phospho-STAT 3 and real-time qPCR for STAT pathway-related genes JAK1 and IL6ST, can reveal upregulated STAT 3. Further demonstration that STAT3 upregulation could cause MEK inhibitor resistance in particular samples may include the use of sh/siRNA against STAT3 in the samples followed by appropriate Western blots for MEK and ERK activation as well as phospho-STAT 3 and total STAT 3. Growth inhibition studies may show that STAT3 knock-down previously sensitizes MEK inhibitor resistant cells to MEK inhibition. Similar effects can be seen if the sample is exposed to a STAT3 inhibitor, such as JSI-124. It was further confirmed that STAT3 upregulation is likely to be responsible for MEK inhibitor resistance in particular tumors from Western blots expressing BIM (including BIM-EL, BIM-L, and BIM-SL). BIM expression leads to MEK inhibitor-induced apoptosis, and STAT3 upregulation therefore reduces BIM levels. STAT3 is known to regulate expression of miR17-92, which inhibits BIM expression. Upregulated STAT3 can lead to higher levels of miR17-92, which will reduce BIM levels and promote resistance to MEK inhibition. Thus, real-time qPCR at miR17-92 levels can also help to assess whether STAT3 upregulation causes MEK inhibition resistance in particular samples. (Dai et al, 2011).

Mutations in the allosteric pocket of MEK that can directly block the binding of inhibitors to MEK or result in constitutive MEK activity can be detected by the methods disclosed below. Emery and colleagues (Emery et al, 2009) and Wang and colleagues (Wang et al, 2011) have previously identified such mutations. Other mutations may affect MEK1 codons located within or adjacent to the N-terminal negative regulatory helix, such as P124L and Q56P. (same as above).

Methods for identifying mutations in nucleic acids (e.g., the MEK genes identified above) are known in the art. Nucleic acids can be obtained from a biological sample. In the present invention, biological samples include, but are not limited to, blood, plasma, urine, skin, saliva, and biopsy samples. The biological sample is obtained from the subject by conventional procedures and methods known in the art.

Non-limiting examples of methods for identifying mutations include PCR, sequencing, hybrid capture, in-solution capture, molecular inversion probes, Fluorescence In Situ Hybridization (FISH) assays, and combinations thereof.

Various sequencing methods are known in the art. These include, but are not limited to, Sanger sequencing (also known as dideoxy sequencing) and various sequencing-by-synthesis (SBS) methods, as disclosed, for example, in Metzker 2005, sequencing by hybridization, sequencing by ligation (e.g., WO 2005021786), sequencing by degradation (e.g., U.S. Pat. nos. 5,622,824 and 6,140,053), and Nanopore sequencing (which is commercially available from Oxford Nanopore Technologies, UK). In deep sequencing techniques, a given nucleotide in a sequence is read multiple times during the sequencing process. Deep sequencing techniques are disclosed, for example, in U.S. patent publication No. 20120264632 and international patent publication No. WO 2012125848.

PCR-based methods for detecting mutations are known in the art and employ PCR amplification, wherein each target sequence in a sample has a corresponding unique pair of sequence-specific primers. For example, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method allows rapid detection of mutations after amplification of genomic sequences by PCR. Mutations were distinguished by digestion with specific restriction endonucleases and identified by electrophoresis. See, for example, Ota et al, 2007. Mutations can also be detected using real-time PCR. See, for example, international application publication No. WO 2012046981.

Hybrid capture methods are known in the art and are disclosed, for example, in U.S. patent publication No. 20130203632 and U.S. patent nos. 8,389,219 and 8,288,520. These methods are based on selective hybridization of a target genomic region to a user-designed oligonucleotide. Hybridization can be to oligonucleotides immobilized on high or low density microarrays (capture on arrays), or to a solution of oligonucleotides modified with a ligand (e.g., biotin), which can then be immobilized to a solid surface, such as a bead (capture in solution).

Molecular Inversion Probe (MIP) technology is known in the art and is disclosed, for example, in Absalan et al, 2008. The method uses MIP molecules, which are specific "padlock" probes for genotyping (Nilsson et al, 1994). MIP molecules are linear oligonucleotides comprising specific regions, universal sequences, restriction sites and Tag (index) sequences (16-22 bp). The MIPs hybridize directly around the target genetic marker/SNP. The MIP method also uses a number of "padlock" probe sets that hybridize in parallel to genomic DNA (Hardenbol et al, 2003). In the case of perfect matches, genomic homology regions are joined by inversion in configuration (as indicated by the technical name) and generation of circular molecules. After the first restriction, all molecules were amplified with universal primers. The amplicons were limited again to ensure short fragments for hybridization on the microarray. The resulting short fragments are tagged and hybridized to the cTag (complementary strand of the index) on the array by Tag sequences. After formation of the Tag-cTag duplex, a signal is detected.

Tables 1, 2 and 3 below show the SEQ ID nos of representative nucleic acid and amino acid sequences of wild-type BRAF, N-RAS and MEK1 from various animals in the sequence listing. These sequences are useful in methods of identifying subjects having mutant BRAF, N-RAS and MEK1 genotypes.

TABLE 1 BRAF sequences

TABLE 2N-RAS sequence

TABLE 3 MEK1 sequence

In another aspect of this embodiment, the method further comprises administering at least one additional therapeutic agent, preferably an inhibitor of the PI3K/Akt pathway, as disclosed herein.

Another embodiment of the invention is a method for treating or ameliorating the effects of cancer in a subject that is refractory or resistant to BRAF inhibitor therapy, MEK inhibitor therapy, or both. The method comprises administering to the subject an effective amount of BVD-523 or a pharmaceutically acceptable salt thereof.

Suitable and preferred subjects are as disclosed herein. In this embodiment, the method can be used to treat the cancers disclosed above, including those having the mutational backgrounds, resistance profiles, and MAPK activities identified above. Methods for identifying such mutations are also described above.

In another aspect of this embodiment, the method further comprises administering to the subject at least one additional therapeutic agent, preferably an inhibitor of the PI3K/Akt pathway, as disclosed herein.

Another embodiment of the invention is a method for identifying a subject with cancer who will benefit from therapy with an ERK inhibitor. The method comprises the following steps:

(a) obtaining a biological sample from a subject; and

(i) the transition between the RAF isoforms is carried out,

(ii) up-regulation of RTK or NRAS signaling,

(iii) reactivation of mitogen-activated protein kinase (MAPK) signaling,

(iv) the presence of a mutation that activates the activation of MEK,

(v) amplification of the mutant BRAF is carried out,

(vi) the STAT3 is up-regulated,

(vii) mutations in the allosteric pocket of MEK that directly block the binding of inhibitors to MEK or that result in constitutive MEK activity,

wherein the presence of the one or more markers confirms that the cancer in the subject is refractory or resistant to BRAF and/or MEK inhibitor therapy, and that the subject would benefit from therapy with an ERK inhibitor that is BVD-523 or a pharmaceutically acceptable salt thereof.

Suitable and preferred subjects are as disclosed herein. In this embodiment, the method can be used to identify subjects with the cancers disclosed above, including those having the mutational backgrounds, resistance profiles, and MAPK activities identified above. Methods for identifying such mutations are also described above.

In one aspect of this embodiment, the method further comprises administering BVD-523 or a pharmaceutically acceptable salt thereof to the subject having one or more of the markers. Preferably, the method further comprises administering to the subject having one or more of the markers at least one additional therapeutic agent, preferably an inhibitor of the PI3K/Akt pathway, as disclosed herein.

Suitable and preferred subjects and types of non-ERK MAPK pathway inhibitor therapy are disclosed herein. In this embodiment, the pharmaceutical composition may be used to treat the cancers disclosed above, including those having the mutational backgrounds, resistance profiles, and MAPK activities identified above. Methods for identifying such mutations are also described above.

In one aspect of this embodiment, the pharmaceutical composition further comprises at least one additional therapeutic agent, preferably an inhibitor of the PI3K/Akt pathway, as disclosed herein.

The kit may also include suitable storage containers, e.g., ampoules, vials, tubes, and the like, for each pharmaceutical composition and other reactant (e.g., buffer, balanced salt solution, and the like) for administration of the pharmaceutical composition to the subject. The pharmaceutical composition and other reactants may be present in the kit in any convenient form, for example in solution or powder form. The kit may further comprise a packaging container, optionally with one or more compartments, for containing the pharmaceutical composition and other optional reagents.

Suitable and preferred subjects and types of non-ERK MAPK pathway inhibitor therapy are disclosed herein. In this embodiment, the kit can be used to treat the cancers disclosed above, including those having the mutational backgrounds, resistance profiles, and MAPK activities identified herein. Methods for identifying such mutations are described above.

In one aspect of this embodiment, the kit further comprises at least one additional therapeutic agent, preferably an inhibitor of the PI3K/Akt pathway, as disclosed herein.

Another embodiment of the invention is a method for inhibiting phosphorylation of RSK in a cancer cell that is refractory or resistant to a non-ERK MAPK pathway inhibitor. The method comprises contacting the cancer cell with an effective amount of BVD-523 or a pharmaceutically acceptable salt thereof for a time sufficient to inhibit RSK phosphorylation in the cancer cell. In this embodiment, "contacting" refers to bringing BVD-523 or a pharmaceutically acceptable salt thereof, and optionally one or more additional therapeutic agents, into close proximity with the cancer cell. This can be achieved using conventional techniques for delivering drugs to mammals, or in an in vitro situation by, for example, providing BVD-523 or a pharmaceutically acceptable salt thereof and optionally other therapeutic agents to the culture medium in which the cancer cells are located. Ex vivo, the contacting may be performed by, for example, providing BVD-523 or a pharmaceutically acceptable salt thereof and optionally other therapeutic agents to the cancer tissue.

Suitable and preferred types of non-ERK MAPK pathway inhibitors are disclosed herein. In this embodiment, affecting cancer cell death can be achieved in cancer cells with various mutational backgrounds, resistance profiles, and MAPK activities as described above. Methods for identifying such mutations are also described above.

The methods of this embodiment, which may be performed in vitro, ex vivo, or in vivo, may be used to effect cancer cell death by, for example, killing cancer cells in cells of the cancer types disclosed herein.

In one aspect of this embodiment, RSK phosphorylation is inhibited by greater than 50%. In another aspect of this embodiment, RSK phosphorylation is inhibited by greater than 75%. In another aspect of this embodiment, RSK phosphorylation is inhibited by greater than 90%. In another aspect of this embodiment, RSK phosphorylation is inhibited by greater than 95%. In another aspect of this embodiment, RSK phosphorylation is inhibited by greater than 99%. In another aspect of this embodiment, RSK phosphorylation is inhibited by 100%.

In another aspect of this embodiment, the cancer cell is a mammalian cancer cell. Preferably, the mammalian cancer cells are obtained from a mammal selected from the group consisting of humans, primates, farm animals, and domestic animals. More preferably, the mammalian cancer cell is a human cancer cell.

In another aspect of this embodiment, the contacting step comprises administering BVD-523 or a pharmaceutically acceptable salt to the subject from which the cancer cells are obtained.

In the present invention, an "effective amount" or "therapeutically effective amount" of a compound or composition disclosed herein is an amount of the compound or composition that, when administered to a subject, is sufficient to produce the beneficial or desired results as described herein. Effective dosage forms, modes of administration, and dosages may be determined empirically, and making such determinations is within the skill of the art. It is understood by those skilled in the art that dosage will vary with the route of administration, rate of excretion, duration of treatment, the nature of any other drug administered, the age, size and species of the mammal (e.g., human patient), and similar factors well known in the medical and veterinary arts. Generally, a suitable dosage of a compound or composition according to the invention will be that amount of such composition which is the lowest dose effective to produce the desired effect. An effective dose of a compound or composition of the invention may be administered as two, three, four, five, six or more sub-doses, administered separately at appropriate intervals throughout the day.

Suitable non-limiting examples of dosages of BVD-523 and other anti-cancer agents disclosed herein are from about 1mg/kg to about 2400mg/kg per day, such as from about 1mg/kg to about 1200mg/kg per day, from 75mg/kg to about 300mg/kg per day, including from about 1mg/kg to about 100mg/kg per day. Other representative dosages of these agents include about 1mg/kg, 5mg/kg, 10mg/kg, 15mg/kg, 20mg/kg, 25mg/kg, 30mg/kg, 35mg/kg, 40mg/kg, 45mg/kg, 50mg/kg, 60mg/kg, 70mg/kg, 75mg/kg, 80mg/kg, 90mg/kg, 100mg/kg, 125mg/kg, 150mg/kg, 175mg/kg, 200mg/kg, 250mg/kg, 300mg/kg, 400mg/kg, 500mg/kg, 600mg/kg, 700mg/kg, 800mg/kg, 900mg/kg, 1000mg/kg, 1100mg/kg, 1200mg/kg, 1300mg/kg, 1400mg/kg, 1500mg/kg, 1600mg/kg, 1700mg/kg, 1800mg/kg, 1900mg/kg, 2000mg/kg, 2100mg/kg, 2200mg/kg and 2300 mg/kg. Effective doses of BVD-523 and other anti-cancer agents disclosed herein may be administered as two, three, four, five, six or more sub-doses, administered separately at appropriate intervals throughout the day.

BVD-523, other inhibitors and various other anti-cancer agents disclosed herein, or the pharmaceutical compositions of the present invention, may be administered in any desired and effective manner: for oral ingestion, or as an ointment or drops for topical administration to the eye, or parenteral or other administration in any suitable manner, e.g., intraperitoneal, subcutaneous, topical, intradermal, inhalation, intrapulmonary, rectal, vaginal, sublingual, intramuscular, intravenous, intraarterial, intrathecal, or intralymphatic. In addition, BVD-523, other inhibitors, and various other anti-cancer agents disclosed herein, or the pharmaceutical compositions of the present invention, may be administered in combination with other therapies. If desired, BVD-523, other inhibitors, and various other anti-cancer agents disclosed herein, or the pharmaceutical compositions of the present invention, may be coated or otherwise protected from the stomach or other secretions.

The pharmaceutical compositions of the present invention comprise one or more active ingredients in admixture with one or more pharmaceutically acceptable diluents or carriers and optionally one or more other compounds, drugs, ingredients and/or materials. Regardless of the route of administration chosen, the agents/compounds of the present invention are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those skilled in the art. See, for example, Remington, The Science and Practice of Pharmacy (21 st edition, Lippincott Williams and Wilkins, philiadelphia, PA).

Pharmaceutically acceptable diluents or carriers are well known in The art (see, e.g., Remington, The science and Practice of Pharmacy (21 st edition, Lippincott Williams and Wilkins, Philadelphia, PA) and The National Formulary (American Pharmaceutical Association, Washington, d.c.) and include sugars (e.g., lactose, sucrose, mannitol, and sorbitol), starch, cellulose preparations, calcium phosphates (e.g., dicalcium phosphate, tricalcium phosphate, and calcium hydrogen phosphate), sodium citrate, water, aqueous solutions (e.g., saline, sodium chloride injection, ringer's injection, dextrose and sodium chloride injection lactide, lactated ringer's injection), alcohols (e.g., ethanol, propanol, and benzyl alcohol), polyols (e.g., glycerol, propylene glycol, and polyethylene glycol), organic esters (e.g., ethyl oleate and triglycerides), biodegradable polymers (e.g., poly-polyethylene glycol), biodegradable polymers (e.g., ethyl oleate and triglycerides), biodegradable polymers (e.g., polyethylene glycol-polyethylene glycol, poly (ortho esters) and poly (anhydrides)), an elastomer base, liposomes, microspheres, oils (e.g., corn, germ, olive, castor, sesame seed and peanut), cocoa butter, waxes (e.g., suppository waxes), paraffins, silicones, talc, silicates, and the like. Each pharmaceutically acceptable diluent or carrier for use in the pharmaceutical compositions of the present invention must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the subject. Diluents or carriers suitable for the selected dosage form and intended route of administration are well known in the art, and acceptable diluents or carriers for the selected dosage form and method of administration can be determined using ordinary skill in the art.

Optionally, the pharmaceutical compositions of the present invention may contain other ingredients and/or materials commonly used in pharmaceutical compositions. Such ingredients and materials are well known in the art and include (1) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (2) binders such as carboxymethyl cellulose, alginate, gelatin, polyvinyl pyrrolidone, hydroxypropyl methyl cellulose, sucrose and acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, sodium starch glycolate, croscarmellose sodium and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorbents, such as quaternary ammonium compounds; (7) wetting agents, such as cetyl alcohol and glycerol monostearate; (8) absorption accelerators such as kaolin and bentonite; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol and sodium lauryl sulfate; (10) suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitol esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth; (11) a buffering agent; (12) excipients, such as lactose, milk sugar (milk sugar), polyethylene glycol, animal and vegetable fats, oils, waxes, paraffins, cocoa butter, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc, salicylates, zinc oxide, aluminum hydroxide, calcium silicate and polyamide powders; (13) inert diluents such as water or other solvents; (14) a preservative; (15) a surfactant; (16) a dispersant; (17) controlled release or absorption delaying agents, such as hydroxypropylmethylcellulose, other polymer matrices, biodegradable polymers, liposomes, microspheres, aluminum monostearate, gelatin and waxes; (18) an opacifying agent; (19) an adjuvant; (20) a wetting agent; (21) emulsifying and suspending agents; (22) solubilizers and emulsifiers such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, oils (in particular cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols, and fatty acid esters of sorbitan; (23) propellants, such as chlorofluorocarbons and volatile unsubstituted hydrocarbons, such as butane and propane; (24) an antioxidant; (25) agents that make the formulation isotonic with the blood of the intended recipient, such as sugars and sodium chloride; (26) a thickener; (27) coating materials, such as lecithin; (28) sweetening agents, flavouring agents, colouring agents, perfuming agents and preserving agents. Each such ingredient or material must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject. Suitable ingredients and materials for the selected dosage form and intended route of administration are well known in the art, and acceptable ingredients and materials for the selected dosage form and method of administration can be determined using ordinary skill in the art.

Pharmaceutical compositions of the invention suitable for oral administration may be in the form of capsules, cachets, pills, tablets, powders, granules, solutions or suspensions of aqueous or non-aqueous liquids, oil-in-water or water-in-oil liquid emulsions, elixirs or syrups, lozenges, boluses, dry lozenges or pastes. These formulations may be prepared by methods known in the art, for example, by conventional pan coating, mixing, granulating, or lyophilizing processes.

Solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules and the like) can be prepared, for example, by mixing the active ingredient with one or more pharmaceutically acceptable diluents or carriers and optionally one or more fillers, extenders, binders, humectants, disintegrating agents, solution retarding agents, absorption enhancers, wetting agents, absorbents, lubricants and/or colorants. Solid compositions of a similar type may be employed as fillers in soft and hard-filled gelatin capsules using suitable excipients. Tablets may be prepared by compression or moulding, optionally containing one or more accessory ingredients. Compressed tablets may be prepared using suitable binders, lubricants, inert diluents, preservatives, disintegrating agents, surface active agents or dispersing agents. Molded tablets may be prepared by molding in a suitable machine. Tablets and other solid dosage forms, such as dragees, capsules, pills, and granules, can optionally be scored or prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may also be formulated to provide slow or controlled release of the active ingredient therein. They may be sterilized by filtration, for example, through a bacteria-retaining filter. These compositions may also optionally contain opacifying agents and may be such that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally in a delayed manner. The active ingredient may also be in the form of microcapsules.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. The liquid dosage form may contain suitable inert diluents commonly used in the art. In addition to inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents. The suspension may contain a suspending agent.

Pharmaceutical compositions of the invention for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more active ingredients with one or more suitable non-irritating diluents or carriers which are solid at room temperature but liquid at body temperature and will therefore melt in the rectum or vaginal cavity and release the active compound. Pharmaceutical compositions of the invention suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such pharmaceutically acceptable diluents or carriers as are known in the art to be appropriate.

Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, drops and inhalants. The active agent/compound may be mixed under sterile conditions with a suitable pharmaceutically acceptable diluent or carrier. Ointments, pastes, creams and gels may contain excipients. Powders and sprays can contain excipients and propellants.

Pharmaceutical compositions of the invention suitable for parenteral administration may comprise one or more agents/compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain suitable antioxidants, buffers, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents. Proper fluidity can be maintained, for example, by the use of coating materials, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. These pharmaceutical compositions may also contain suitable adjuvants such as wetting agents, emulsifying agents and dispersing agents. It may also be desirable to include isotonic agents. Furthermore, the absorption of the injectable pharmaceutical form may be prolonged by the inclusion of an agent that delays absorption.

In some cases, in order to prolong the effect of a drug (e.g., a pharmaceutical formulation), it is desirable to slow its absorption from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material which is poorly water soluble.

The rate of absorption of the active agent/drug then depends on its rate of dissolution, which in turn depends on crystal size and crystal form. Alternatively, delayed absorption of the parenterally administered agent/drug may be achieved by dissolving or suspending the active agent/drug in an oily vehicle. Injectable depot forms can be prepared by forming a matrix of microcapsules of the active ingredient in a biodegradable polymer. Depending on the ratio of active ingredient to polymer and the nature of the particular polymer used, the rate of release of the active ingredient can be controlled. Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues. The injectable material may be sterilized, for example, by filtration through a bacterial-retaining filter.

The formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried condition requiring only the addition of the sterile liquid diluent or carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the type described above.

The present invention provides treatment of cancers that are refractory or resistant to non-ERK MAPK pathway inhibitor therapy and discloses combinations that are shown to potentiate the effects of ERK inhibitors. In this context, the applicant has also shown that combinations of different ERK inhibitors also have a synergistic effect. Thus, it is expected that the effect of the combinations described herein may be further improved by the use of one or more additional ERK inhibitors. Accordingly, some embodiments of the invention include one or more additional ERK inhibitors.

The invention also provides a method of treating a subject with unresectable or metastatic BRAF600 mutation positive melanoma comprising administering to the subject 600mg BID of BVD-523 or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the mutation is BRAF^V600EAnd (4) mutation.

The present invention also provides a composition comprising 600mg BVD-523 or a pharmaceutically acceptable salt thereof and optionally a pharmaceutically acceptable carrier, adjuvant or vehicle for use in the treatment of a subject with unresectable or metastatic BRAF600 mutation positive melanoma.

The following examples are provided to further illustrate the process of the present invention. These examples are illustrative only and are not intended to limit the scope of the present invention in any way.

Examples

Example 1

Materials and methods

Cancer cell lines were maintained in cell culture under standard media and serum conditions. For dose escalation studies, a375 cells were split, grown to approximately 40-60% confluence, and then treated with the initial dose of the specified drug. Table 4 shows a summary of the incremental drug treatments.

TABLE 4 summary of incremental processing

Single reagent dose escalation was performed based on Little et al, 2011 and is summarized in fig. 20. Cells were then grown until 70-90% were pooled and distributed. The split ratio remained as "normal" as possible and reasonably consistent between treatments (e.g., a minimum of 50% of the normal split ratio of the parent). The medium was refreshed every 3-4 days. When the cells again reach about 40-60% confluence, the dose is escalated. If a window of 40-60% is missed, the cells are again split and dosing begins once they reach 40-60% confluence. Similarly, the medium was refreshed every 3-4 days. The process is repeated as necessary (fig. 20).

For single agent treatment, the initial concentration and dose increase were performed as follows: to approximate IC₅₀Initially, the dose is doubled in small increments, or slightly, for the first 4-5 doses, increased in the same increments for the next 4 doses, and then the concentration of subsequent doses increased by 1.5 times.

For the combined treatment, the initial concentrations and dose increases were performed as follows: at about IC of each compound₅₀Beginning at half (the combined assay indicates that this would result in an inhibition range of about 40-70%), increasing by a single agent (i.e., making an initial doubling, then increasing the same increment in the next 4 doses, then a 1.5-fold increase in concentration). Table 5 shows the expected dose increase using these regimens.

TABLE 5 predicted dose escalation-month one

Clonal resistant cell populations are derived from resistant cell banks by limiting dilution.

Changes in sensitivity to the increasing agent are followed at appropriate time intervals (e.g., monthly, although time depends on the sufficient number of cells available) using a proliferation assay. For proliferation assays, cells were seeded at 3000 cells per well in 96-well plates in drug-free DMEM media containing 10% FBS and allowed to adhere overnight before compound or vehicle controls were added. Compounds were prepared from DMSO stocks to give final concentration ranges as shown in figures 2A-2H. The final DMSO concentration was constant at 0.1%. Test compounds were incubated with cells at 37 ℃ and 5% CO₂Incubate for 96 hours in a humid atmosphere. Alamar blue 10% (v/v) was then added and incubated for 4 hours and the fluorescent products were detected using a BMG FLUOstar plate reader. Mean medium-only background values were derived and the data were analyzed using the 4-parameter logistic equation in GraphPad Prism. Paclitaxel was used as a positive control.

Proliferation assays at month 1 beginning on day 28 using cells grown at the concentrations of each agent shown in table 6.

TABLE 6 initial concentration of drug used in proliferation assay-month 1

Cell lines	Dab	Tram	BVD-523
				Parent strain	-	-	-
Tram	-	2nM	-
				Dab	15nM	-	-
BVD-523	-	-	0.48μM
				Tram+Dab	5nM	1nM	-
Dab+BVD-523	7.5nM	-	0.24μM
				Tram+BVD-523	-	1nM	0.16μM

Proliferation assays at month 2 beginning on day 56 were performed using cells grown at the concentrations of each agent shown in table 7.

TABLE 7 initial concentration of drug used in proliferation assay-month 2

Cell lines	Dab	Tram	BVD-523
				Parent strain	-	-	-
Tram	-	8nM	-
				Dab	127nM	-	-
BVD-523	-	-	0.8μM
				Tram+Dab	10nM	2nM	-
Dab+BVD-523	12.5nM	-	0.4μM
				Tram+BVD-523	-	2nM	0.32μM

At the end of the 3 month increment period, cultures were maintained at the highest concentration for 2 weeks prior to the last round of proliferation assay and possible single cell cloning. Since the proliferation assay/single cell cloning requires actively proliferating cells, the spare culture was also kept at a lower concentration for treatments where cells proliferate very slowly at the highest concentration or only recently increased (table 8). For BVD-523 treatment, where the cells appear to stop growing almost completely and appear particularly fragile at the highest concentration (1.8. mu.M), the culture was maintained at lower concentrations for a period of 2 weeks.

TABLE 8 treatment details of 2 weeks incubation at fixed concentration

The proliferation assay at month 3 used cells grown at the concentrations of each agent shown in table 9.

TABLE 9 initial concentration of drug used in proliferation assay-month 3

Cell lines	Dab	Tram	BVD-523
				Parent strain	-	-	-
Tram	-	160nM	-
				Dab	3.2μM	-	-
BVD-523	-	-	1.2μM
				Tram+Dab	80nM	16nM	-
Dab+BVD-523	28nM	-	0.9μM
				Tram+BVD-523	-	2.5nM	0.4μM

For the combination studies, a375 cells (ATCC) were seeded at a cell density of 3000 cells/well in triplicate 96-well plates in DMEM plus 10% FBS and allowed to adhere overnight before the test compound or vehicle control was added. The combination was tested using a 10x8 dose matrix with a final DMSO concentration of 0.2%. Followed by a 96 hour assay incubation period, followed by the addition of Alamar blue 10% (v/v) and incubation for 4 hours, followed by reading on a fluorescence plate reader. After reading Alamar blue, the medium/Alamar blue mixture was flicked and 100. mu.l of CellTiter-Glo/PBS (1:1) was added and the plates were treated according to the manufacturer's (Promega) instructions. Media background alone values were subtracted prior to analysis of the data. The Bliss adduction model was then applied.

Briefly, equation C is used_blissThe predicted fractional inhibition values for the combined inhibition were calculated as a + B- (a × B), where a and B are the fractional inhibition obtained for drug a alone or drug B alone at a specific concentration. If the combination of the two drugs is exactly additive, C_blissIs the expected fractional inhibition. Subtraction of C from the experimentally observed fractional inhibition value_blissValue, give an 'excess over Bliss' value. An excess over Bliss of greater than 0 indicates synergy, while values less than 0 indicate antagonism. Excess over Bliss was plotted as heat map ± SD.

Single and combined data are also presented as dose-response curves generated in GraphPad Prism (plotted using% viability relative to DMSO-only treated control).

For the combination study of interest, the Alamar blue viability assay was performed as described above for the combination study. Alternatively, a Caspase-Glo 3/7 assay was performed. Briefly, HCT116 cells were seeded in white 96-well plates in McCoy's5A plus 10% FBS in triplicate at a cell density of 5000 cells/well. A375 cells were seeded at 5000 cells/well in DMEM plus 10% FBS. Cells were allowed to adhere overnight before the test compound or vehicle control was added. The final concentration of DMSO was 0.2%, and 800nM staurosporine was included as a positive control. Assay incubation periods of 24 and 48 hours were used. Then, Caspase-3/750% (v/v), the plates were mixed on an orbital shaker for 5 minutes and incubated at room temperature for 1 hour before reading on a plate reader. Media background alone values were subtracted prior to analysis of the data.

For differential scanning fluorometry, SYPRO orange (5,000 Xsolution, Invitrogen) was diluted (1:1,000) in buffer solution (10mM HEPES, 150mM NaCl, pH 7.5) to (1:1,000). HisX6 tagged proteins including inactive ERK2, active ERK2(ppeRK2) or p38 α at a final concentration of 1 μ M. Compounds in protein/dye solution and 100% DMSO were added to the wells (2% v/v final DMSO concentration) to reach the desired final concentration, mixed and placed in an RT-PCR instrument.

For the Ki assay of ERK1, activated ERK1(10nM) was combined with various concentrations of compounds in 2.5% (v/v) DMSO at 30 ℃ in 0.1M HEPES buffer (pH 7.5), 10mM MgCl₂2.5mM phosphoenolpyruvate, 200. mu.M Nicotinamide Adenine Dinucleotide (NADH), 150. mu.g/mL pyruvate kinase, 50. mu.g/mL lactate dehydrogenase and 200. mu.M Erktide peptide for 10 minutes. The reaction was initiated by the addition of 65 μ M ATP. Monitoring the reduced absorbance (340nm) and determining the IC₅₀As inhibitorsAs a function of formulation concentration.

For the Ki determination of ERK2, the inhibitory activity of BVD-523 on ERK2 was determined by radiometric assay, with final concentrations of 100mM HEPES (pH 7.5), 10mM MgCl₂1mM Dithiothreitol (DTT), 0.12nM ERK2, 10. mu.M Myelin Basic Protein (MBP) and 50. mu.M³³P-gamma-ATP. All reaction components except ATP and MBP were premixed and aliquoted (33 μ Ι _) into 96-well plates. Stock solutions of compounds in DMSO were used to prepare up to 500-fold dilutions; to each well was added a 1.5 μ L aliquot of DMSO or inhibitor in DMSO. By adding substrate³³The reaction was initiated with P-gamma-ATP and MBP (33. mu.L). After 20 min, the reaction was quenched with 20% (w/v) trichloroacetic acid (TCA) (55. mu.L) containing 4mM ATP, transferred to GF/B filter plates and washed 3 times with 5% (w/v) TCA. Add Ultimate Gold^TMAfter scintillation with 50. mu.L of scintillator, the samples were counted in a PackardTopcount. Ki values were determined from activity versus concentration titration curves by fitting the data to an equation of competitive tight binding inhibition kinetics using Prism software version 3.0.

IC for ERK2₅₀Assay, activity was determined by standard coupled enzyme assay. The final concentrations were as follows: 0.1MHEPES (pH 7.5), 10mM MgCl₂1mM DTT, 2.5mM phosphoenolpyruvate, 200. mu.M NADH, 50. mu.g/mL pyruvate kinase, 10. mu.g/mL lactate dehydrogenase, 65. mu.M ATP, and 800. mu.M peptide (ATGPLSPGPFGRR). All reaction components except ATP were premixed with ERK and aliquoted into assay plate wells. BVD-523 in DMSO was introduced into each well, keeping the DMSO concentration constant per well. The BVD-523 concentration ranged from 500 times per drop. The assay plate was incubated in the plate reading chamber of a spectrophotometer (molecular device) for 10 minutes at 30 ℃ and then the reaction was initiated by the addition of ATP. Monitoring the change in absorbance at 340nm as a function of time; the initial slope corresponds to the reaction rate. Fitting the rate versus concentration of the BVD-523 titration curve to the equation of competitive tight binding inhibition kinetics to determine Ki values or 3 parameter fitting to determine IC using Prism software version 3.0₅₀。

For apoptosis assays, cells are plated in individual wells2×10⁴Individual cells were seeded in 96-well plates and allowed to adhere overnight or grown to 50% confluence. Cells were treated with serial dilutions of BVD-523 in culture medium (final volume 200. mu.L, concentration range 4-0.25. mu.M) and CO at 37 ℃₂Incubate in incubator for 48 hours. Cells were washed with 100. mu.L of PBS and 60. mu.L of radioimmunoprecipitation assay buffer (50mM Tris-HCl, pH 8.0, 150mM NaCl, 1.0% [ w/v ]]NP-40，0.5％[w/v]1% of sodium deoxycholate [ w/v ]]SDS), and then incubated at 4 ℃ for 10 minutes to lyse the cells. A30- μ L aliquot of the lysate was added to 100 μ L of caspase assay buffer (120mM HEPES, 12mM EDTA, 20mM dithiothreitol, 12.5 μ g/mL AC-DEVD-AMC caspase substrate) and incubated at room temperature for 4 hours to overnight. The plates were read in a fluorimeter (excitation wavelength 360nm, emission wavelength 460 mm). The remaining 30 μ L of lysate was analyzed for total protein content using the BioRad protein assay kit (sample to working reagent ratio of 1: 8). The final normalized caspase activity was derived as fluorescent units per μ g protein and converted to a fold increase in caspase activity compared to DMSO control.

To measure anti-tumor activity in a375 xenografts, xenografts were initiated with a375 cells maintained by serial subcutaneous transplantation in female athymic nude mice. Each test mouse received a375 tumor fragments (1 mm) subcutaneously implanted on the right side³). Once the tumor reaches the target size (80-120 mm)³) Animals were randomly assigned to treatment and control groups and drug treatment was initiated.

To evaluate BVD-523 monotherapy, BVD-523 in 1% (w/v) carboxymethylcellulose (CMC) was administered orally (p.o.) at BID doses of 5, 25, 50, 100, or 150 mg/kg. Temozolomide was administered orally as a positive reference compound at 75 or 175mg/kg once daily (QD) for a total of five treatments (QD × 5).

The efficacy of BVD-523 in combination with dabrafenib was evaluated in mice randomized into 9 groups of 15 and 1 group of 10 (i.e. group 10). Oral administration of dabrafenib at 50 or 100mg/kg QD and oral administration of BVD-523 at 50 or 100mg/kg BID, alone and in groupsAnd (4) merging until the study is finished; vehicle-treated and temozolomide-treated (150mg/kg QD × 5) controls were also included. Combined dosing was stopped on day 20 to monitor tumor regrowth. Animals were monitored individually and when each tumor reached 2000mm³At the end-point volume, or on the last day (day 45) (whichever comes first), euthanasia was performed and the median Time To End (TTE) was calculated. The combination was also evaluated in the late A375 model, where 228-³Larger tumors in the range. Here, the mice were randomly divided into 1 group of 14 (group 1) and 4 groups of 20 (groups 2 to 5). Administration was initiated on day 1 with dabrafenib plus BVD-523(25mg/kg dabrafenib +50mg/kg BVD-523 or 50mg/kg dabrafenib +100mg/kg BVD-523), with BID being administered orally for each agent until the end of the study. The study included a 50-mg/kg dabrafil and 100-mg/kg BVD-523 monotherapy group and a vehicle treated control group. Tumors were measured twice weekly. Combined dosing was stopped on day 42 to monitor tumor regrowth to the end of the study (day 60). Treatment outcome was determined by% TGD, defined as the percentage increase in median TTE of treated versus control mice, with differences between groups analyzed by log rank survival analysis. For TGI analysis, the initial (i) and final (f) tumor measurements based on the following formulas were used to calculate and report% TGI values for each treatment (T) group versus control (C): % TGI ═ 1-Tf-Ti/Cf-C. Mice were also monitored for CR and PR responses. Animals with CR at the end of the study were additionally classified as TFS.

To measure BVD-523 activity in Colo205 xenografts, human Colo205 cells were cultured in RPMI 1640 supplemented with 10% (v/v) Fetal Bovine Serum (FBS), 100 units/mL penicillin, 100 μ g/mL streptomycin (Invitrogen) and 2mM L-glutamine. Cells were cultured for less than 4 passages prior to implantation. On day 0, female athymic nude mice (19-23g) were injected subcutaneously with 2X10⁶Individual Colo205 cells were directed to the right dorsal axillary region.

Will have an approximate tumor volume of 200mm³The mice were randomly divided into 6 experimental groups. A vehicle control, i.e., 1% CMC (w/v), was prepared weekly. Suspending BVD-523 in 1% (w/v) CMC at the desired concentrationAnd homogenized on ice at 6,500rpm for 50 minutes. BVD-523 suspensions were prepared weekly and administered orally with BID at total daily doses of 50, 100, 150 and 200mg/kg (n-12/group) on an 8 or 16 hour dosing schedule for 13 days. Vehicle control (n-12) was administered using the same dosing regimen. CPT-11 was administered as a positive reference compound (n-12). Each 1mL of CPT-11 injection contained 20mg of irinotecan, 45mg of sorbitol, and 0.9mg of lactic acid. CPT-11 was administered intraperitoneally every 4 days at 100 mg/kg/day for 2 consecutive doses.

To measure the ERK1/2 isotopically-tagged internal standard (ITIS) mass spectra in Colo205 xenografts, frozen tumors were lysed in 10 volumes of ice-cold lysis buffer (10mM TRIS-HCl, pH 8.0, 10mM MgCl₂，1％(v/v)Triton X-100，Complete^TMProtease inhibitor cocktail [ Roche, Cat # 1836170]Phosphatase inhibitor cocktail I [ Sigma, cat # P-2850]Phosphatase inhibitor cocktail II Sigma Cat No. 5726]And benzonase [ Novagen Cat No. 70664]) And (4) cracking. The lysate was clarified by centrifugation (100,000 Xg, 4 ℃, 60 min) and the supernatant adjusted to 2 mg/mL with lysis buffer. ERK1 was immunoprecipitated using agarose coupling and pan-anti-ERK 1(Santa Cruz Biotechnology Cat. No. sc-93ac) antibody. Immunoprecipitated proteins were resolved by SDS-PAGE and stained with SYPRO Ruby (Invitrogen), and ERK bands were excised by razor. Gel sections were cut at 300. mu.L of 20mM NH₄HCO₃Washed, cut into small pieces, and placed in a Page Eraser Tip (The New Group catalog number SEM 0007). The gel fragments were reduced and alkylated prior to trypsinization. The tryptic fraction was separated in 75. mu.L of 50% (v/v) acetonitrile, 0.2% (v/v) trifluoroacetic acid and the resulting sample was concentrated to 0-10. mu.L in a SpeedVac.

For ITIS analysis, digested samples were spiked with atom-tagged peptide standards and fractionated phosphorylation was quantified by coupled liquid chromatography-tandem Mass Spectrometry (MS). Using a Rheos 2000 binary pump from Flux Instruments (providing nanoscale flow after 1:750 min.), an LC packing insetsil nano pre-packed column (C18, 5mm,30mm ID. times.1 mm) and a New Objective PicoFrit AQUASIL analytical column (C18, 5mm, 75/15mm ID. times.10 cm) (which can also be used as an electrospray ionization (ESI) emitter) were used for nanocapillary chromatography. An Applied Biosystem API 3000 mass spectrometer coupled to a nano ESI source was used for MS analysis. An internally fabricated gas nozzle connected to an atomizing gas source is used to help stabilize the nanoflow spray. Data were obtained in a Multiple Reaction Monitoring (MRM) mode: the atomizing gas is 3; the curtain gas is 7; the collision gas is 5; the ion injection voltage is 2150V, and the outlet potential is 10V; Q1/Q3 resolution is low/unit; and the dwell time for all MRM channels is 65 milliseconds. All raw MS data was processed using a combination of the analysis software suite from Applied biosystems and custom tools.

To evaluate drug sensitivity in the cell line model for acquired resistance, drug sensitivity was evaluated in 96 hour proliferation assays for increasing doses of a375 cells and isogenic RKO cells. RKO isogenic cells (containing 10% [ v/v% ]]McCoy's5A for FBS) or dose-escalated a375 cells (DMEM containing 10% FBS) were seeded into 96-well plates and allowed to adhere overnight before compound or vehicle controls were added. Note that increasing doses of a375 cells were seeded in the absence of inhibitor. Compounds were prepared from 0.1% (v/v) DMSO stock to give the final concentrations indicated. Test compounds were incubated with cells at 37 ℃ in 5% CO₂Incubation was carried out in a humid atmosphere for 96 hours. For RKO cells, CellTiter-Reagents (Promega) and luminescence detected using a BMG FLUOstar plate reader. For the A375 assay, Alamar blue (ThermoFisher) 10% (v/v) was added and incubated for 4 hours, followed by detection of fluorescent products using BMG FLUOstar. The average medium-only background values were subtracted and the data were analyzed using the 4-parameter logistic equation in GraphPad Prism.

IC of ERK1 was measured in a final reaction volume of 25. mu.L₅₀And (4) measuring. ERK1 (human) (5-10mU) and25mM Tris (pH 7.5), 0.02mM ethylene glycol tetraacetic acid, 250. mu.M peptide, 10mM magnesium acetate and gamma-³³P-ATP (specific activity approximately 500cpm/pmol, concentrated as required) was incubated together. The reaction was initiated by the addition of Mg ATP. After 40 min incubation at Room Temperature (RT), the reaction was stopped by adding 5. mu.L of a 3% (w/v) phosphoric acid solution. Then, 10 μ L of the reaction was spotted on a P30 filter pad, washed 3 times in 75mM phosphoric acid for 5 minutes, then once in methanol, then dried and scintillation counted.

RKO MEK1Q56P isogenic cells were generated from Horizon Discovery (Cambridge, UK; # HD106-019) using a recombinant AAV-mediated gene targeting strategy. Briefly, rAAV viruses were produced after transfection of appropriate targeting and helper vectors in HEK293T cells, purified using AAV purification kit (Virapur, San Diego, USA) and titrated using qPCR. Parental homozygous RKO cells (homozygous wild type for MEK 1) were then infected with rAAV virus and clones that had integrated the selection cassette were identified by G418 selection and amplified. Correctly targeted clones were identified by PCR and sequencing, which were heterozygous for the MEK1Q56P point mutation knock-in single isogene.

Isogenic SW48 cell lines heterozygous for the knock-in mutant KRAS (De Roock et al, 2010, JAMA, 304, 1812-. For the proliferation assay, cells were seeded into 96-well plates in McCoy's5A medium supplemented with 10% FBS and allowed to adhere overnight before compound or vehicle controls were added. Test compounds were incubated with cells at 37 ℃ in 5% CO₂Incubate under atmosphere for 96 hours. Viability was then assessed using Alamar blue.

A proprietary KinaseProfiler assay was performed at Upstate Discovery and a radioactivity assay similar to that used by Davies et al was used to describe the selectivity of BVD-523 for a panel of 70 kinases.

Drug sensitivity assays were performed as part of the drug sensitivity genomics in cancer projects using high throughput screening as previously described (Yang et al, 2013).

For Western blot analysis, A375 cells were seeded onto 10cm dishes in Dulbecco's modified Eagle's medium plus 10% (v/v) FBS. Cells were allowed to adhere overnight before the test compound or vehicle was added. For experiments using RKO cells, these cells were seeded in 6-well plates or 10cm dishes with McCoy's5A + 10% (v/v) FBS. The cells are then treated at the desired concentration and duration. Cells were harvested by trypsinization, pelleted and snap frozen. Lysates were prepared with RIPA buffer supplemented with a mixture of protease and phosphatase inhibitors (Roche), clarified by centrifugation at 11,000rpm for 10 minutes, and quantified by the bicinchoninic acid assay. Samples were resolved by SDS-PAGE, blotted onto polyvinylidene difluoride membranes and probed with antibodies against the indicated targets (i.e., pRB [ Ser780], Cat No. 9307; CCND1, Cat No. ab 6152; BCL-xL, Cat No. 2762; PARP, Cat No. 9542; DUSP6, Cat No. 3058S).

For Reverse Phase Protein Assay (RPPA), A375, MIAPaCa-2, HCT116, Colo205, HT-29 and AN3Ca cells (ATCC) were plated at 80% confluence, allowed to recover overnight (MIAPaCa-2 cells plated at 30% confluence and allowed to recover for 3 days), and then treated with 10 μ M of each compound (i.e., BVD-523, SCH722984, GDC-0994 or Vx-11e) for 6 hours at 37 ℃. Control wells were treated with 0.1% (v/v) DMSO for 6 hours prior to generation of cell lysates. The samples were then analyzed using reverse phase protein microarray technology (Theranostics Health).

For analysis of pERK IHC in Colo205 xenografts, xenograft tumors were treated overnight in 70% to 100% graded ethanol, washed in two changes of xylene, infiltrated with paraffin, and embedded in paraffin blocks. Then, 5- μm sections were cut and placed on positively charged glass slides and baked at 60 ℃ for at least 30 minutes, but not longer than 1 hour. Individual sections from each animal and dose group were probed with an anti-phospho-p 42/p44MAPK antibody (pERK [1:100], CST; Cat. No. 9101; batch No. 16), counterstained with hematoxylin, and then microscopically analyzed using a Zeiss Axioplan 2 microscope. Isotype controls (rabbits, Zymed laboratories, cat # 08-6199, lot # 40186458) were used as negative controls.

For FACS analysis, cells were scraped and pelleted at 1,500rpm for 5 minutes, then resuspended in 1mL of buffer and frozen at-70 ℃. The frozen cells were thawed and centrifuged again, then resuspended in 0.25mL of buffer a (trypsin in spermine tetrahydrochloride decontamination buffer) for 10 minutes to disaggregate the cell mass and digest the cell membrane and cytoskeleton. Buffer B (trypsin inhibitor and ribonuclease I in buffer, 0.2mL) was added for 10 minutes in the dark. The resulting DNA stained nuclei were filtered and analyzed by FACS. Based on the presence of n and 2n DNA (or higher) levels, the histograms were analyzed to determine the proportion of cells in the G1, S and G2/M phases of the cell cycle.

To measure in vitro combinatorial activity, five thousand G-361 cells were seeded into triplicate 96-well plates containing McCoy's5A and 10% (v/v) FBS and allowed to adhere overnight. The vemurafenib/BVD-523 combination was tested using a 10x8 dose matrix. The compounds were incubated with cells at 37 ℃ in 5% CO₂Incubate in a humid atmosphere for 72 hours. CellTiter-Glo reagent was added according to the manufacturer's instructions and luminescence was detected using an MBG FLUOstar plate reader. Dose matrix interactions were determined by Loewe additivity and Bliss independence models using Horizon's Chalice combinatorial analysis software.

For compound resistance to be developed in vitro by dose escalation, A375 parent cells (ATCC CRL-1619) were grown to-40-60% confluence in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% heat-inactivated FBS and penicillin/streptomycin, and then treated with initial doses of BVD-523, trametinib, or dabrafenib, either alone or in combination, with IC for each compound₅₀Or slightly lower than the IC of each compound₅₀The concentration of (c); for combination studies, the initial dose was the IC of each compound₅₀Half of that. Growing the cells until about 70-90% are pooled and distributed; the medium was refreshed every 3-4 days. When the cells again reach about 40-60% confluence, the doses were performed in the same increments (equal to the starting concentration)Increasing, then increasing the concentration 1.5 fold, then further increasing by 2 fold if the cells continue to adapt rapidly. (e.g., the first six doses of ascending dabrafenib are: 5, 10, 15, 20, 25, and 37.5 nM). The process is repeated as necessary.

The cell viability assay of fig. 30A was performed by resazurin (Alamar blue) metabolic assay after 5 days in drug in whole serum under high glucose conditions. Cells were seeded at approximately 15% -50% confluence in 384 well microtiter plates in medium containing 10% FBS and penicillin/streptavidin plus glucose (18-25 mM). The optimal number of cells for each cell line was determined to optimize growth during drug administration. For adherent cell lines, after overnight incubation, cells were treated with 9 concentrations of each compound (2-fold dilution series) using a liquid handling robot and returned to the incubator for assay at 96 hour time points. For suspension cell lines, cells were treated with compound immediately after plating and returned to the incubator for a 96 hour time point. Cells were then stained with 55 μ g/ml resazurin (Sigma) prepared in glutathione free medium for 4 hours. For Resazurin, the fluorescence signal intensity was quantified using a fluorescence plate reader at an excitation and emission wavelength of 535/595 nm. Strict quality control measures were performed on all screening plates. The effect on cell viability was measured and a curve fitting algorithm was applied to the raw data set to obtain a multi-parameter description of drug response, including half maximal Inhibitory Concentration (IC)₅₀)。IC₅₀IC in μ M₅₀Natural logarithm of (LN _ IC)₅₀(ii) a EXP Return to IC in μ M₅₀). Allowing extrapolation of IC in cases where very high values are produced₅₀. By bonding the IC if necessary₅₀Values were limited to the maximum test concentration (and low values of the minimum test concentration), limiting the data to the range of concentrations tested.

Efficacy testing for BVD-523 in patient-derived xenografts (AT052C) representative of those from BRAF that have become clinically refractory to vemurafenib^V600EMelanoma in the patient. Tumor fragments were harvested from host animals and implanted into immunodeficient mice. TheThe mean tumor volume at the start of the study was approximately 170mm³At this point, animals were randomized into 4 groups, including the control group (1% [ v/v ])]Oral administration of CMC BID X31) and 3 treatment groups (BVD-523[100mg/kg ]]Dalafinil [50 mg/kg)]Or BVD-523/dabrafenib [100/50mg/kg]N is 10/group); all therapeutic agents were administered orally and on a BID X31 schedule.

IC for inhibition of PMA stimulated RSK1 phosphorylation by BVD-523 in human whole blood samples₅₀Determination of IC of BVD-523 inhibiting PMA-stimulated RSK1 phosphorylation in 10 healthy donors (age 22-61 years)₅₀Values, where an 8-point concentration curve was used, ranged from 10 μ M to 5nM BVD-523. The control consisted of 3 unstimulated and 3 PMA-stimulated samples per donor. phosphorus-RSK (pRSK) and total RSK levels were determined and data were calculated using pRSK/RSK levels for each sample.

30 ml of blood was drawn from each donor into a heparin sodium evacuated blood collection tube. To each of 22 2-mL microtubes per donor was added 1mL of whole blood. Microtubules are labeled with donor numbers (1 to 10) and subsequent treatment names: "A" for PMA stimulation only (maximal), "B" for BVD-523-containing samples receiving PMA stimulation; and "C" indicates an unstimulated sample (minimum). Dimethyl sulfoxide (DMSO) was added to all tubes of groups a and C to a final concentration of 0.1%. The sample was then gently shaken at room temperature.

BVD-523(10mM in 100% DMSO) was serially diluted 3-fold into 100% DMSO. These serially diluted BVD-523 samples in 100% DMSO were then diluted 10-fold in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum and penicillin/streptomycin/glutamine, and 10. mu.L of each of these working solutions was added per mL of blood for each of the indicated BVD-523 concentrations. BVD-523 at each concentration was run in duplicate, with two 1mL blood samples each, to give 16 total samples for a complete 8-point concentration curve. The sample was then gently shaken at room temperature for at least 2 hours but no longer than 3 hours.

Human whole blood samples in groups a and B of all donors were stimulated with PMA at a final concentration of 100nM for 20 min at room temperature. The samples in group C were not treated with PMA but were shaken and treated as all other samples.

After completion of PMA treatment of each sample, peripheral blood mononuclear cells were isolated from human whole blood. 1mL of blood from each sample was gently spread onto 0.75mL of room temperature Histopaque 1077 in a 2mL microcentrifuge tube. The samples were centrifuged in an Eppendorf microcentrifuge for 2 minutes at 16,000 Xg. The interface and upper layer were removed and added to a tube containing 1mL of cold Dulbecco's Phosphate Buffered Saline (DPBS). These samples were then centrifuged at 16,000 Xg for 30 seconds to pellet the cells. The buffer supernatant was removed by aspiration and the pellet was resuspended in 1mL of cold DPBS. The pellet from each sample was then re-pelleted as described above. The buffer was removed by aspiration and the pellet was lysed as shown below.

Complete lysis buffer consisted of Meso Scale Discovery Tris lysis buffer, 1 xhalt protease inhibitor cocktail, 1X phosphatase inhibitor cocktail 2, 1X phosphatase inhibitor cocktail 3, 2mM phenylmethanesulfonyl fluoride and 0.1% sodium dodecyl sulfate. The lysis buffer was kept on ice and each sample set was freshly prepared. The final cell pellet was lysed by adding 120 μ L of complete lysis buffer. The sample was vortexed until the cell pellet disappeared and then snap frozen on dry ice. Samples were stored at-20 ℃ before measurement of pRSK and total RSK by ELISA.

For pRSK ELISA (PathScan), thawed lysates were combined with sample diluent (provided in ELISA kit) in a 1:1 combination: mu.L of lysate was added to 120. mu.L of sample diluent in a round bottom 96 well plate. This combination was then transferred to pRSK microwells at 100 μ Ι _ per well. For total RSK ELISA (PathScan), 20 μ L of lysate that had been diluted 1:1 in sample diluent was further diluted in 200 μ L of sample diluent in a round bottom 96 well plate. The combination was then transferred to all RSK microwells at 100 μ L per well. The plates were sealed with plate seals and incubated at 4 ℃ for 16 to 18 hours, which was the time shown to produce the best detection of the target protein. Both ELISAs were developed according to the kit instructions.

If the patient has incurable, histologically confirmed metastatic or advanced malignancy; ECOG performance status is 0 or 1; adequate kidney, liver, bone marrow and heart function; and life expectancy ≥ 3 months, then a patient of ≥ 18 years of age is eligible for participation. Patients may receive up to 2 antegrade chemotherapies for their metastatic disease. Exclusion criteria were known uncontrolled brain metastases; gastrointestinal disorders that may impair study drug absorption; a history or current signs/risk of retinal vein occlusion or central serous retinopathy; and concurrent therapy with drugs known as CYP1a2, strong inhibitors of CYP2D6 and CYP3a4, or strong inducers of CYP3a 4. All participants provided informed consent prior to the initiation of any study procedure.

Patients receiving at least one dose of BVD-523 were included in the analysis using SAS (version 9.3) software. The expiration date is 2016, 12, month, and 1. This study was registered in clinical trials. gov, and was numbered NCT 01781429.

The present invention provides data from an open label, multicenter phase I study to evaluate the safety, pharmacokinetics and pharmacodynamics of increasing doses of BVD-523 in patients with advanced malignancies. The dosing regimen was combined with an accelerated titration and a standard cohort 3+3 dose escalation regimen, which were used together to identify the MTD and RP2D of BVD-523 in patients with advanced solid tumors. Between 1 and 6 patients in each treatment cohort were assigned to receive higher oral doses of BVD-523 in the 21 day cycle in order of the BID schedule (12 hour interval), starting with a dose of 10mg BID. BVD-523 was administered continuously BID over a 21 day period at the following doses: 10mg (n ═ 1); 20mg (n ═ 1); 40mg (n ═ 1); 75mg (n-1); 150mg (n ═ 1); 300mg (n-4); 600mg (n-7); 750mg (n-4); and 900mg (n ═ 7).

Patients received oral doses of BID until disease progression, unacceptable toxicity or clinical observation that met another exit criterion. Dose escalation in individual patient groups occurred in up to 100% increments until 1 patient experienced grade 2 toxicity (not including alopecia or diarrhea). The cohort was then expanded to at least 3 patients per cohort, with subsequent dose escalation increments decreasing from up to 100% to up to 50%. When at least 1 patient in the 3 patient cohort experienced DLT, up to 3 additional patients were treated at this dose level. When more than 1 DLT occurs in ≦ 6 patients, the dose level is defined as the non-tolerated dose and dose escalation is stopped. Patient internal dose escalation is allowed, provided that the patient receiving the highest current dose has been observed for at least 3 weeks, and that less than 2 of 6 patients at a given dose are reported for dose limiting side effects. Patients experiencing DLT or unacceptable toxicity discontinue their treatment until toxicity returns to grade ≦ 1. Recovery of BVD-523 treatment was then initiated at the lower dose level of the next test or at a 20% to 30% dose reduction (compared to capsule dose).

The primary objective of the phase I study was to determine the safety and tolerability of BVD-523 by determining dose-limiting toxicity, MTD and RP 2D. Secondary goals include the determination of the pharmacokinetic profile of BVD-523 in patients with advanced malignancies, as well as studies to assess any preliminary clinical effects on tumor response by physical or radiological examination using RECIST v 1.1. The exploration objective includes evaluating pharmacodynamic marker (biomarker) measurements and studies¹⁸Preliminary clinical effects of F-FDG-PET assessment on tumor response, as shown.

To determine MTD, DLT and RP2D, MTD was defined as the highest dose cohort, where < 33% of patients experienced a BVD-523-associated DLT in the first 21 days of treatment. DLT is defined as BVD-related toxicity at 21 days prior to treatment, resulting in grade 4 hematological toxicity lasting >1 day; grade 3 hematologic toxicity with complications (e.g., thrombocytopenia with bleeding); grade 3 non-hematologic toxicities, except untreated nausea, vomiting, constipation, pain, and rash (these become DLT if AE persists despite adequate treatment); or treatment was discontinued for more than 3 days during cycle 1 (or cycle 2 was not entered for >7 days) due to BVD-523-associated toxicity.

RP2D may be as high as the MTD and is determined in discussions with clinical researchers, medical monitors, and sponsors. Included in the rationale underlying RP2D are observations related to pharmacokinetics, pharmacodynamics, and any cumulative toxicity observed after multiple cycles.

With respect to safety assessments, AE is defined as any adverse medical event in a patient administered a pharmaceutical product that is not necessarily causal to BVD-523 and is encoded using the MedDRA encoding dictionary. SAE is any adverse medical event that occurs at any dose that results in death, life-threatening, requiring hospitalization or prolonging existing hospitalization, or that results in persistent or significant disability/disability or congenital abnormalities/birth defects. The severity of AE was graded according to the national cancer institute adverse event general term criteria, scoring scale, version 4.

Safety assessments were made at baseline on days 8, 15, 22, 29, 36, and 43, and every 3 weeks or after if clinically indicated in patients who continued treatment. Each evaluation included physical examination and clinical laboratory studies. The electrocardiogram is repeated if clinically significant and at the discretion of the investigator. Researchers judge whether an AE is relevant to the study drug and follow up until resolution or stability, or the AE is judged to no longer be clinically significant.

For pharmacokinetic analysis, the pharmacokinetic population consisted of patients receiving at least one dose of BVD-523 and having evaluable pharmacokinetic data for plasma and/or urine. Blood samples were then collected prior to dosing at 0.5 (+ -5 min), 1 (+ -5 min), 2 (+ -10 min), 4 (+ -10 min), 6 (+ -10 min), 8 (+ -10) and 12 (+ -2 h) hours after morning dose on day 1 (visit 2; baseline/treatment initiation) and day 15 (visit 4; steady state). On day 22, prior to dose administration, final blood samples were collected for pharmacokinetic analysis. Urine samples were collected on day 1 and day 15 at intervals of 1 to 6 hours and 6 to 12 ± 2 hours before and after dosing. Plasma and urine samples were analyzed for BVD-523 and metabolites using a validated LC/MS/MS method. Standard pharmacokinetic parameters were obtained in a non-compartmental manner using Phoenix WinNonlin (Pharsight). The relationship between dose and exposure was calculated using standard least squares regression analysis.

For pharmacodynamic confirmation of BVD-523 against target inhibition, targeted ERK inhibition of BVD-523 was determined by detecting pRSK as a target biomarker in human whole blood samples obtained from advanced solid tumor patients (N-27) who received different doses of BVD-523(10-900mg BID) during the phase I study. The activity of BVD-523 was expressed as the percentage activity of PMA-stimulated blood (pRSK) incubated with BVD-523 from 4 time points (pre-baseline, 4 hours post-baseline, pre-15 and 4 hours post-15 days).

To measure anti-tumor response, tumor measurements based on physical examination occurred at baseline and on the first day of each treatment cycle. CT and other evaluations were performed every 2 to 3 cycles. Evaluation results according to RECIST v 1.1: CR is defined as the disappearance of all target lesions; PR is defined as the total reduction in the longest diameter of the target lesion of greater than or equal to 30% with reference to the baseline measurement; stable disease is defined as neither sufficient contractile competence for PR nor sufficient increased competence as progressive disease, measured at baseline. Before the first BVD-523 dose and on day 15 (visit 4)¹⁸F-FDG-PET scanning for tumor observation¹⁸F-glucose uptake was used to assess metabolic response.

Example 2

Dose escalation and proliferation assay-first month

Dose escalation progression-first month

A375 cells were dose-escalated using BVD-523, dabrafenib and trametinib as single agents or combinations. During the first month, the dose is increased in small increments. In addition to the significant decrease in growth rate, the cells generally tolerated the dose escalation well, and are scheduled to escalate the dose more aggressively at month 2 using larger increments. Figures 1A-1C show the 1 st month progression of the dose escalation study.

Proliferation assay results-first month

Proliferation assays were performed to assess the response of the incremental cell lines versus the parental cell line to BVD-523, dabrafenib, and trametinib treatments.

Figures 2A-2H show the results of the normalized and original proliferation assay from month 1 of the study. Note that the difference in maximum signal in DMSO control between different treatments (fig. 2D, 2F, and 2H) indicates differential growth rates between treatments. These differences may affect the response of the cell line to the inhibitor in the proliferation assay.

Table 10 shows the IC at month 1 of the study₅₀And (4) data.

TABLE 10-IC₅₀Data-month 1

Par ═ parental cell line

It was earlier suggested that cells grown in the presence of dabrafenib or trametinib, whether as single agents or in combination, at increasing doses, exhibited a reduced response to both agents in the proliferation assay.

In the early stage of month 2, the growth rate of cells in the dabrafenib-only treatment was significantly increased relative to the early stage of month 1. This increased the rate of progression and indicated that resistance became apparent.

Example 3

Dose escalation and proliferation assay-second month

Dose escalation progression-second month

The second month of the study found that most of the treatments entered the initial mild escalation phaseThe phase in which the dose is increased in larger increments (1.5 fold). The single agents of dabrafenib and trametinib increased most rapidly, and the concentration of cell growth corresponded to 100x parental cell IC₅₀(FIG. 3A and FIG. 3B). The single agent of BVD-523 progressed more slowly than dabrafenib and trametinib (FIG. 3C). See figure 3D for comparison of single agent increments. The increased BVD-523 cells had a more "fragile" appearance and there were a greater number of floating cells compared to the increasing populations of dabrafenib and trametinib.

The combined agent delivery progressed more slowly than the single agent treatment. The BVD-523/trametinib combination is particularly effective in preventing cell progression.

Proliferation assay results-second month

Proliferation assays of single agent-escalated dabrafenib and trametinib cell populations show modest changes in dose response curves, suggesting that additional escalation periods would be advantageous for further enrichment of resistant cells. Interestingly, in the proliferation assay, there is evidence that cells exposed to BVD-523 do not grow well after inhibitor withdrawal, which may indicate an addictive level.

Fig. 4A-4H show the results of the normalized and original proliferation assay from month 2 of the study. Note that the difference in maximum signal in DMSO control between different treatments (fig. 4D, 4F, and 4H) indicates differential growth rates between treatments. These differences may affect the response of the cell line to the inhibitor in the proliferation assay.

Fig. 5A-5H show the results of the normalized and raw proliferation assay from month 2 of the study, focusing only on the parental and BVD-523 line data.

Table 11 shows the IC at month 2 of the study₅₀And (4) data. Relative IC₅₀Determined by 4-parameter curve fitting in Prism.

TABLE 11-IC₅₀Data-month 2

Par ═ parental cell line

Example 4

Dose escalation and proliferation assay-month three

Dose escalation progression-third month

Figures 6A-6C show single and combined agent escalation at month 3 of the study. Figure 6D shows a comparison of single agent increments.

Proliferation assay results-month three

Figure 7 shows growth assessment during proliferation assay in DMSO control wells. Fig. 8A-8D show the results of the study at month 3. FIGS. 9A-9D show the results of month 3 of the study, focusing on single treatment cell lines.

Table 12 shows the IC at month 3 of the study₅₀And (4) data. Relative IC₅₀Determined by 4-parameter curve fitting in Prism. The IC of cell lines that were escalated with trametinib was not determined due to lack of growth during the assay₅₀Value (ND: not performed).

TABLE 12 IC₅₀Data-month 3

Par ═ parental cell line

Figure 19 shows single and combination agent escalation at month 3 of the study. The obtained cellsLineage variants can have greater IC of these agents in dabrafenib or trametinib than in parental a375 cells₅₀In the presence of a concentration 100 times higher than that of the growth medium. In contrast, cell lines resistant to BVD-523 can only be kept lower than the parental IC₅₀At a concentration 10 times higher. Sensitivity tests indicate that the dalafinib or trametinib resistant cell line remains relatively sensitive to BVD-523; IC of BVD-523 in resistant cell lines₅₀"Shift" increasing ratio of corresponding IC after Dalafinil or trametinib treatment₅₀The increase is more moderate. Likewise, when compared to dabrafenib or trametinib treatment, the IC concentration is its IC in the parental a375 line₅₀When a 10-fold higher BVD-523 treatment of resistant cell lines was performed, more complete inhibition of cell growth was observed. Overall, the pattern of resistance and cross-sensitivity suggests that BVD-523 may remain effective in the case of acquired resistance.

Example 5

Combined results of the study

As expected, a375 cells carrying the BRAF (V600E) mutation were sensitive to dabrafenib. Single reagent IC calculated using Alamar blue compared to those derived using CellTiter-Glo (FIGS. 11A-11E, 13A-13E, and 15A-15E)₅₀The values (fig. 10A-10E, 12A-12E, and 14A-14E) are typically slightly lower for dabrafenib and BVD-523. Published IC of dabrafenib and trametinib in a 72-hour CellTiter-Glo assay₅₀Values were 28 + -16 nM and 5 + -3 nM, respectively (Greger et al, 2012; King et al, 2013) -single agent results reported herein are consistent with these values. There is some evidence that all treatments have a window of synergy. The variation between triplicates was small, however, there was some evidence of marginal effects that might explain the significantly enhanced growth observed in some treatments versus the no-drug control (e.g., particularly evident in the trametinib/BVD-523 combination). This makes interpretation of the Bliss analysis more challenging, as it may lead to artificial enhancement of the level of synergy in certain treatments.

The combinatorial assay was repeated on a375 cells. The single agent BVD-523, trametinib and dabrafenib potency are consistent with those reported in previous studies disclosed herein.

Taken together, the combined data show that MEK and BRAF resistant cells can be overcome by treatment with the ERK inhibitor BVD-523.

Example 6

Marker and effector function of BVD-523 altering MARK kinase Activity

For Western blot studies, HCT116 cells (5X 10)⁶) The cells were plated in 10cm dishes in McCoy's5A plus 10% FBS. A375 cells (2.5X 10)⁶) The cells were plated in 10cm dishes in DMEM plus 10% FBS. Cells were allowed to adhere overnight before the indicated amount of test compound (BVD-523) or vehicle control was added. Prior to isolation of whole cell protein lysates, cells were treated for 4 or 24 hours, as described below. Cells were harvested by trypsinization, pelleted and snap frozen. Lysates were prepared with RIPA (radioimmunoprecipitation assay) buffer, clarified by centrifugation and quantified by a bicinchoninic acid assay (BCA) assay. 20-50 μ g of protein was resolved by SDS-PAGE electrophoresis, blotted onto PVDF membrane and probed using antibodies detailed in Table 13 (for 4 hour treatment) and Table 14 (for 24 hour treatment) below.

TABLE 13 details of the antibodies

TABLE 14 details of antibodies

Fig. 16A-16D, 17A-17D, and 18A-18B show Western blot analysis of cells treated with BVD-523 at various concentrations as follows: 1) MAPK signaling component in a375 cells after 4 hours; 2) periodic and apoptotic signaling in 24-hour a375 cells treated with varying amounts of BVD-523; 3) MAPK signaling in HCT-116 cells treated for 4 hours. The results indicate that both urgent and long-term treatment of RAF and RAS mutant cancer cells with BVD-523 in vitro affect both substrate phosphorylation and effector targets of ERK kinase. The BVD-523 concentration required to induce these changes is typically in the low micromolar range.

Several variations of specific active markers are notable. First, the abundance of the slow-migrating isoform of ERK kinase increased after BVD-523 treatment; moderate changes can be observed acutely and can be increased after prolonged treatment. Although this may indicate an increase in enzymatically active, phosphorylated form of ERK, it is still noteworthy that various proteins directly and indirectly regulated by ERK remain "off" after BVD-523 treatment. First, the RSK1/2 protein shows reduced phosphorylation at residues that are strictly dependent on ERK for protein modification (T359/S363). Secondly, BVD-523 treatment induced a complex change in MAPK feedback phosphatase DUSP 6: slow migrating protein isoforms decreased after the emergency treatment, while total protein levels were greatly reduced after the long-term BVD-523 treatment. These two findings are consistent with reduced activity of ERK kinase, which controls DUSP6 function through post-translational and transcriptional mechanisms. Overall, although the cellular form of ERK that is generally considered active is increased, it appears that cellular ERK enzyme activity may be completely inhibited following an emergency or long-term treatment with BVD-523.

Consistent with these observations, changes in effector genes requiring MAPK pathway signaling were required following treatment with BVD-523. The G1/S cell cycle organelle (appaatus) is regulated at the post-translational and transcriptional levels by MAPK signaling and cyclin-D1 protein levels are greatly reduced after prolonged BVD-523 treatment. Similarly, gene expression and protein abundance of apoptotic effectors generally required intact MAPK signaling and the total level of Bim-EL increased after prolonged BVD-523 treatment. However, as described above, PARP protein lysis and increased apoptosis were not noted in the a375 cell background; this suggests that other factors may influence whether changes in BVD-523/ERK dependent effector signaling translate into deterministic events such as cell death and cell cycle arrest.

Consistent with the cellular activity of BVD-523, marker analysis indicates that ERK inhibition alters multiple molecular signaling events in cancer cells, rendering them susceptible to both reduced cell proliferation and survival.

In summary, fig. 16A-16D, 17A-17D, and 18A-18D show that BVD-523 inhibits the MAPK signaling pathway, and may be more favorable compared to RAF or MEK inhibition in this case.

Finally, the nature of BVD-523 may make it a preferred agent for use as an ERK inhibitor compared to other agents with similar activity. Kinase inhibitor drugs are known to show unique and specific interactions with their enzyme targets, and drug efficacy is strongly influenced by the direct inhibition pattern and susceptibility to adaptive changes that occur after treatment. For example, inhibitors of ABL, KIT, EGFR and ALK kinases are only effective when their cognate targets are present in either an active or inactive configuration. Also, some of these inhibitors are uniquely sensitive to secondary gene mutations or post-translational adaptive changes in protein targets. Finally, RAF inhibitors show different potency towards RAF kinases present in certain protein complexes and/or subcellular localisation. In summary, since ERK kinases are similarly known to exist in a wide variety of, variable and complex biochemical states, it seems likely that BVD-523 might interact with and inhibit these targets in a different and highly preferred manner than other agents.

Example 7

Effect of BVD-523 and baseline ERK BRAF and MEK inhibitors on viability and MARK signaling

Single reagent proliferation assay

Cells were seeded in 96-well plates at the densities shown in table 15, in McCoy's5A containing 10% FBS, and allowed to adhere overnight before compound or vehicle controls were added. Compounds were prepared from DMSO stocks to give the desired final concentrations. The final DMSO concentration was constant at 0.1%. Test compounds were incubated with cells at 37 ℃ and 5% CO₂Incubate for 96 hours in a humid atmosphere. Addition of CellTiter-Reagents (Promega, Madison, WI) and detection of luminescence using a BMG FLUOstar plate reader (BMG Labtech, Ortenberg, Germany). The mean medium-only background values were subtracted and the data were analyzed using a 4-parameter logistic equation in GraphPad Prism (GraphPad Software, La Jolla, CA).

Combinatorial proliferation assays

Cells were seeded in triplicate 96-well plates at the densities indicated in table 15, in McCoy's5A containing 10% FBS, and allowed to adhere overnight before the test compound or vehicle control was added. The combinations were tested using a 10x8 dose matrix. The final DMSO concentration was constant at 0.2%.

Test compounds were CO-administered with cells at 37 ℃ and 5% CO₂Incubate for 96 hours in a humid atmosphere. Cells were stained with Hoechst staining and fluorescence was detected as described above. The average medium-only background values were subtracted and the data analyzed.

Using Chalice^TMCombined analysis software (Horizon Discovery Group, Cambridg)e, MA), the combined interaction of the dose matrices is determined by Loewe additivity and Bliss independence models, as described in the user manual (available in the book. horizontal and coverage. com/book-port/documentation/analyzer/home. jsp). Synergy was determined by comparing experimentally observed inhibition levels for each combination point with the expected value of additivity, derived from single agent responses along the matrix edges. Potential synergistic interactions were identified by showing calculated excess inhibition over what was predicted to be additive in the dose matrix as a heat map, and by quantifying the 'synergy score' based on the Loewe model report. Single reagent data from a combinatorial assay plate are presented as in Charice^TMThe dose-response curve generated in (a).

TABLE 15 cell line seeding Density

Western blot

Cells were seeded in 6-well plates (experiment 1) or 10cm petri dishes (experiment 2) at the densities shown in table 15, in McCoy's5A containing 10% FBS, and allowed to adhere overnight before compound or vehicle controls were added. Test compounds were added and CO-incubated with cells at 37 ℃ and 5% CO₂Incubate in humid atmosphere for 4 or 24 hours. Cells were harvested by trypsinization, pelleted by centrifugation and snap frozen on dry ice.

Lysates were prepared using RIPA buffer (50mM Tris-hydrochloride, pH 8.0; 150mM sodium chloride; 1.0% Igepal CA-630 (NP-40); 0.5% sodium deoxycholate; 0.1% sodium dodecyl sulfate; 1 × complete EDTA-free protease inhibitor cocktail (Roche, Nutley, NJ; cat 05892791001); 1xphos STOP phosphatase inhibitor cocktail (Roche Nutley, NJ; cat.04683907001)) and clarified by centrifugation at 11,000rpm for 10 minutes in a tabletop centrifuge.

According to the manufacturer's instructions (Pierce)^TMBCA ProteinAssay Kit; thermo Scientific, Waltham, MA; catalog No. 23225) quantitated total protein in the lysate by BCA assay, in sample buffer (NuPAGE LDS sample buffer; (Invitrogen, Carlsbad, CA; cat. NP0007)) and stored at-80 ℃.

Equal amounts of protein (40. mu.g) were resolved on NuPAGE 4-12% Bis-Tris gels (Invitrogen, Carlsbad, Calif.; Cat. No. WG1402BOX) and blotted onto PVDF membrane using an iBlot gel transfer stack (Invitrogen, Carlsbad, Calif.; Cat. No. IB4010-01) on an iBlot gel transfer apparatus (Invitrogen Carlsbad, Calif.) according to the manufacturer's instructions.

The blots were probed using the antibodies and blocking conditions detailed in table 16. Use of Pierce^TMWestern blot substrate ECL2 Western blot (Thermo Scientific, Waltham, MA; Cat. No. 80196) Western blots were developed and imaged using a FluorChem MWestern blot imager (ProteinSimple, San Jose, Calif.).

TABLE 16 antibodies and Western blot conditions

The MEK1(Q56P) mutation exemplifies a class of clinically relevant MEK1/2 activating mutations known to upregulate the MAPK pathway and drive acquired resistance to BRAF or MEK inhibitors.

The study used a pair of RKO BRAF (V600E) cell lines, isogenic to the presence or absence of the MEK1(Q56P) activation mutation, to evaluate the effect of the activating MEK mutation in response to the novel ERK inhibitor BVD-523 versus other benchmark MAPK inhibitors.

By using CellTiter-Cellular ATP levels were quantified to assess the effect on cell viability. Single reagent assays demonstrated that double mutant BRAF (V600E) inhibition of MEK1(Q56P) cells against baseline clinical BRAF (e.g., dabrafenib) or MEK (e.g., trametinib) inhibitors showed significantly reduced sensitivity relative to parental BRAF (V600E) cells, demonstrating that this isogenic model is useful for clinically reproducing the acquired resistance known to be associated with such mutations (table 17).

TABLE 17 Single reagent IC₅₀Value of

n.d. ═ not determined, only partial dose responses were obtained

In contrast, the response to BVD-523 was identical in both parental and double mutant cells, indicating that BVD-523 is insensitive to this mechanism of acquired resistance.

These results were identical in two independently derived double mutant BRAF (V600E): MEK1(Q56P) cell line clones, confirming that these differences in response to the control parental cells were specifically correlated with the presence of the MEK1 mutation and not with unrelated clone artifacts (fig. 22A-fig. 22E). Similar results were also observed with a second mechanistically different baseline ERK inhibitor (SCH772984), supporting the notion that these observations are specifically related to inhibition of ERK rather than due to off-target effects.

Charice from Horizon using the Loewe additive or Bliss independent model^TMCombinatorial analysis software, the effect of BVD-523 in combination with BRAF inhibitors (e.g. dabrafenib) was also assessed across the concentration matrix in these cell lines (fig. 23-fig. 23O and fig. 24A-fig. 24O). Calculated excess inhibition was then shown to exceed that expected for the dose matrix as a heatmap as additive, and the 'volume fraction' was calculated to show whether the overall response to the combination was synergistic (positive), antagonistic (negative) or additiveIs evaluated for the presence of potential synergistic interactions.

The results show that the BVD-523: Dalanib combination is mainly additive in both the parental and mutant cell lines. In contrast, the combination of MEK inhibitor (trametinib) garrafenib, although mainly additive in the parental cell line, showed strong synergy in the double mutant BRAF (V600E): MEK1(Q56P) cell line (fig. 25A-fig. 25O). The Loewe volume, Bliss volume and synergy score for the test combinations are shown in tables 18-20, respectively, and are shown graphically in fig. 26A-26C.

TABLE 18 Loewe volume

TABLE 19 Bliss volume

TABLE 20 synergistic Scoring

Significant effects on the MAPK pathway were assessed by Western blotting. Basal ERK phosphorylation levels (DMSO samples) were significantly upregulated in the MEK1(Q56P) expression line relative to the parent, further confirming that this isogenic model faithfully recapitulates the expected phenotype of expression of the MEK activation acquired resistance mutation.

In parental BRAF (V600E) RKO cells, a reduced level of RSK1/2 phosphorylation was observed following an emergency treatment with pharmacologically active concentrations of RAF, MEK and ERK kinase inhibitors. In contrast, the isogenic double mutant BRAFV600E, MEK1Q56P cells did not show reduced RSK phosphorylation after BRAF or MEK inhibitor treatment, while BVD-523 was still effective at similar concentrations (FIG. 27A-FIG. 27I). The dashed line indicates that the trametinib-treated sample (plus the matched DMSO control) and the blot are from separate experiments on BRAFi and BVD-523-treated samples.

Following prolonged inhibitor treatment, changes in effector gene signaling consistent with the cytostatic pattern were observed. In parental RKO lines, a decrease in phosphorylated pRB levels was observed following prolonged MEK and ERK inhibitor treatment. At pRB regulatory levels, the MEK1 mutant system appears to be insensitive to treatment with low concentrations of MEK inhibitor, while higher concentrations remain effective. Crucially, the efficacy of BVD-523 against pRB activity did not appear to be strongly affected by MEK mutations. Surprisingly, RAF inhibitor treatment did not affect pRB status despite effective inhibition of upstream signaling in the parental and MEK mutant context.

Taken together, these results show that BVD-523 is not susceptible to MEK activation mutations, such as MEK1(Q56P), driven acquired resistance. Furthermore, they propose that the interaction between BVD-523 and BRAFi (e.g. dabrafenib) in the combination is additive regardless of the presence or absence of MEK activating mutations.

Example 8

Combinatorial interactions between ERK inhibitors

RAF mutant melanoma cell line a375 cells were cultured in DMEM containing 10% FBS and seeded into triplicate 96-well plates at an initial density of 2000 cells per well. The combined interaction between the ERK inhibitors BVD-523 and SCH772984 was analyzed after 72 hours as described in example 4 above. CellTiter-Reactivity (Promega, Madison, Wis.) was assayed and BMG FLUOstar was usedLuminescence was detected by a plate reader (BMGLAbtech, Ortenberg, Germany).

Visualization of the Loewe and Bliss 'excess inhibition' heat maps indicated that the combination of BVD-523 and SCH772984 was predominantly additive and had windows of potential synergy in the mid-range dose (fig. 28A-28E).

Taken together, these results indicate that the interaction between BVD-523 and SCH772984 is at least additive and in some cases synergistic.

Example 9

Targeting the MAPK signaling pathway in cancer: promising activity using a novel selective ERK1/2 inhibitor BVD-523(Ulixertinib)

The therapeutic strategies for cancer have evolved from classical cytotoxicity-based approaches to agents that counteract the effects of genetic foci that drive aberrant signaling essential to tumor proliferation and survival. For example, in addition to the constitutively active mutations of pathway components such as RAS and BRAF (Gollob et al, 2006), the ERK module of the mitogen-activated protein kinase (MAPK) signaling cascade (RAS-RAF-MEK-ERK) (Cargnello and Rouxx 2011) may be involved in several receptor tyrosine kinases (e.g., EGFR and ErbB-2). Genetic alterations in RAS or BRAF lead to rapid tumor growth, increased cell survival and resistance to apoptosis through aberrant activation of ERK signaling (pourikoks et al, 2011, Corcoran et al, 2010, Nazarian et al, 2010, Shi et al, 2014, Wagle et al, 2011). Activating mutations in RAS family members KRAS and NRAS account for approximately 30% of all human cancers, with particularly high incidence in pancreatic (Kanda et al, 2012) and colorectal (Arrington et al, 2014) cancers. Constitutively activating mutations in the BRAF gene normally encoding valine at amino acid 600 are observed in melanoma, thyroid cancer, colorectal cancer, and non-small cell lung cancer (Hall et al, 2014). Cancers that carry genetic mutations that result in changes in the downstream components ERK and MEK have also been reported (Ojesina et al, 2014, Arcila et al, 2015). Alterations that activate the MAPK pathway are also common in the case of resistance to targeted therapies (Groenendijk et al, 2014). Thus, targeting the MAPK pathway terminal major kinase (ERK1/2) is a promising strategy for tumors with this pathway activation alteration (e.g., BRAF, NRAS, and KRAS).

Three MAPK pathway-targeted drugs have been approved by the united states Food and Drug Administration (FDA) for single agent therapy with BRAF^V600Mutant unresectable or metastatic cutaneous melanoma: the BRAF inhibitors vemurafenib and dabrafenib and the MEK inhibitor trametinib. In addition, a combination of dabrafenib and trametinib was also approved for this indication (queiro et al, 2015 and Massey et al, 2015). Additional MEK inhibitor cobimetinib (cobimetinib) was approved for use in this indication as part of the combination regimen with BRAF inhibitor. Clinical experience with these drugs has demonstrated that the MAPK pathway is a therapeutic target. In BRAF^V600In phase III trials of mutant melanoma patients, the single agents vemurafenib and dabrafenib showed excellent response rates (approximately 50% versus 5-19%) and median progression-free survival (PFS, 5.1-5.3 months versus 1.6-2.7 months) over cytotoxic chemotherapy (dacarbazine) (Chapman et al, 2011 and Hauschild et al, 2012). Furthermore, clinical applications with BRAF-plus MEK targeting therapy have demonstrated that simultaneous targeting of different nodes in the MAPK pathway can enhance the magnitude and duration of response. The first-line use of BRAF plus MEK-targeting agents (dabrafenib/trametinib or cobitinib/vemurafenib) further improved median overall survival compared to single agent BRAF inhibition (Robert et al, 2015, Long et al, 2015, Larkin et al, 2014). Thus, for having BRAF^V600Mutant metastatic melanoma patients, combined BRAF-/MEK-targeted therapy is a valuable treatment option.

Although improvements in clinical outcome were observed with the BRAF-/MEK-inhibitor combination therapy, the long-lasting benefit was limited by acquired resistance and subsequent eventual development of disease progression, with a median PFS range of about 9-11 months. (Robert et al, 2015, Long et al 2015, Larkin et al 2014 andand Flaherty et al, 2012). The genetic mechanism of acquired resistance to inhibition by a single agent, BRAF, has been extensively studied and resistance mechanisms identified to include splice variants of BRAF (pourikakos et al, 2011), BRAF^V600EAmplification (Corcoran et al, 2010), MEK mutation (Wagle et al, 2014), NRAS mutation and RTK activation (Nazarian et al, 2010 and Shi et al, 2014). Resistance mechanisms in the case of BRAF-/MEK-inhibitor combination therapies began to emerge and reflected the mechanism of BRAF single agent resistance (Wagle et al, 2014 and Long et al, 2014). These genetic events share the ability to reactivate ERK signaling. Indeed, reactivated MAPK pathway signaling measured by ERK transcriptional targets is common in tumor biopsies from BRAF inhibitor resistant patients (Rizos et al, 2014). Furthermore, ERK1/2 reactivation is observed without a resistance inheritance mechanism (Carlino et al, 2015). Thus, seeking to achieve sustained clinical benefit has led researchers to focus on evaluating other agents that target the downstream MAPK component, ERK 1/2. Inhibition of ERK may provide important clinical benefit to patients with acquired resistance to BRAF/MEK inhibition. ERK family kinases have shown promise as therapeutic targets in preclinical cancer models, including those resistant to BRAF or MEK inhibitors (Morris et al, 2013 and hatzivasssiliou et al, 2012). However, the potential use of this ERK1/2 inhibitor extends beyond the acquired resistance of melanoma.

Targeting ERK1/2 is a rational strategy in any tumor type carrying a known driver of MAPK, not only in patients with relapse BRAF/MEK treatment. Since ERK1 and ERK2 are located downstream of this pathway, they represent a particularly attractive therapeutic strategy in the MAPK cascade, which can avoid upstream resistance mechanisms. Herein, preclinical features of BVD-523(ulixertinib) in MAPK pathway dependent cancer models including drug first treatment and BRAF/MEK therapy acquired resistance models are reported. The results of the phase I dose discovery study of BVD-523 are included as a companion publication to this journal. See examples 17-24.

In the present invention, BVD-523 is shown to be an effective, highly selective, reversible small molecule ATP competitive inhibitor of ERK1/2 with in vitro and in vivo anti-cancer activity.

BVD-523 (ulixentinib) was identified and characterized as a novel reversible ATP-competitive ERK1/2 inhibitor with high efficacy and ERK1/2 selectivity. BVD-523 caused a reduction in proliferation and an increase in caspase activity, most pronounced in cells carrying MAPK (RAS-RAF-MEK) pathway mutations. In vivo BRAF^V600EIn xenograft studies, BVD-523 showed dose-dependent growth inhibition and tumor regression. Interestingly, BVD-523 inhibited phosphorylation of the target substrate, despite increased phosphorylation of ERK 1/2. BVD-523 also exhibits anti-tumor activity in an acquired resistance model to single agent and combination BRAF/MEK targeted therapies. When BVD-523 is used in combination with BRAF inhibition, BRAF has also been demonstrated^V600E-synergistic antiproliferative effect in a xenograft model of a mutant melanoma cell line. These studies suggest that BVD-523 is promising as a therapeutic approach for ERK-dependent cancers, including those in which the tumor is acquired resistant to other therapies targeting nodes upstream of the MAPK pathway.

Example 10

Discovery and preliminary characterization of novel ERK1/2 inhibitor BVD-523(Ulixertinib)

Following extensive optimization of the initially identified clues using high-throughput small molecule screening (Aronov et al, 2009), a novel Adenosine Triphosphate (ATP) competitive ERK1/2 inhibitor BVD-523(ulixertinib) was identified (fig. 29A). BVD-523 is a potent ERK inhibitor with a Ki of 0.04. + -. 0.02nM for ERK 2. It appears to be a reversible competitive inhibitor of ATP because of the IC of ERK2 inhibition₅₀Values increased linearly with increasing ATP concentration (fig. 29B and 29C). For incubation times of 10 minutes or more, IC₅₀It remained almost constant, indicating a rapid equilibrium and association of BVD-523 with ERK2 (FIG. 29D). BVD-523 is also a tight binding inhibitor of recombinant ERK1 (Rudolph et al, 2015) showing Ki<0.3nM。

Binding of BVD-523 to ERK2 was demonstrated using calorimetric studies and compared to data generated using the ERK inhibitors SCH772984 and pyrazolyl pyrrole (Arovov et al, 2007). all compounds bind and stabilize inactive ERK2 at increased concentrations as shown by positive Δ Tm values (fig. 29E). the 10 to 15 degree change in Δ Tm observed using BVD-523 and SCH-772984 is consistent with compounds with low nanomolar binding affinity (Fedorov et al, 2012). BVD-523 shows strong binding affinity to phosphorylated active ERK2(pERK2) and inactive ERK2 (fig. 29F). stronger affinity to rk2 is observed compared to inactive ERK 2. BVD-523 does not interact with negative control protein p38 α MAP kinase (fig. 29F).

In addition to ERK1 and ERK2, BVD-523 showed excellent ERK1/2 kinase selectivity based on a biochemical counter screen against 75 kinases. ATP concentration approximately equal to K in all assays_m. Kinases inhibited by greater than 50% by 2 μ M BVD-523 were retested to produce K_iValue (or apparent K)_i(ii) a Table 21). 12 of the 14 kinases have<K of 1. mu.M_i. The selectivity of BVD-523 to ERK2, with the exception of ERK1, was for all kinases tested>7000 fold Ki with inhibition of ERK1<0.3nM (10 fold). BVD-523 is therefore a highly potent and selective inhibitor of ERK 1/2.

Table 21-BVD-523 shows selectivity for ERK1 and ERK2 kinases.

Example 11

BVD-523 preferentially inhibits cell proliferation and enhances caspase-3/7 activity in vitro in cancer cell lines with MAPK pathway activating mutations

BVD-523 cell activity was evaluated in a set of about 1,000 cancer cell lines of different lineages and genetic background (fig. 30A and table 22). Cell lines are classified as MAPK wild-type (wt) or mutant according to the absence or presence of mutations in RAS family members and BRAF. Although some MAPK-wt cell lines are sensitive to BVD-523, BVD-523 typically preferentially inhibits proliferation in cells with MAPK pathway alterations.

Next, the effect of BVD-523 treatment on the growth and survival of sensitive cells was characterized. After 24 hours of treatment with BVD-523 at 500nM or 2000nM, in BRAF^V600EFluorescence Activated Cell Sorting (FACS) analysis on the mutant melanoma cell line UACC-62. The treated cells arrested in a concentration-dependent manner in the G1 phase of the cell cycle (fig. 30B).

In addition, caspase-3/7 activity was analyzed as a measure of apoptosis in various human cancer cell lines. Concentration and cell line dependent increase of caspase 3/7 was observed after 72 hours of treatment with BVD-523 (FIG. 30C). BVD-523 treatment in carrier BRAF^V600The mutant MAPK activated cell line subset (a375, WM266 and LS411N) resulted in significant caspase-3/7 induction. This is consistent with previous observations regarding preferential inhibition of BVD-523 proliferation in MAPK pathway-mutant cancer cell lines (fig. 30A).

To further characterize the mechanism of action and effect on BVD-523-induced signaling, BRAF treated at BVD-523^V600ELevels of various effector proteins and MAPK-related proteins were assessed in mutant a375 melanoma cells (fig. 30D). After 4 and 24 hours of BVD-523 treatment, phospho-ERK 1/2 levels increased in a concentration-dependent manner. Although an outstanding concentration-dependent increase in pERK1/2 was observed with treatment with 2. mu. MBVD-523, phosphorylation of the ERK1/2 target RSK1/2 decreased at both 4 and 24 hours, consistent with sustained inhibition. Total protein levels of DUSP 6(a distant marker of ERK1/2 activity) also attenuated at 4 and 24 hours. After 24 hours of treatment with BVD-523, the apoptosis marker BIM-EL increased in a dose-dependent manner, while cyclin D-1 and pRB decreased at 2. mu.M. All effects are consistent with targeted ERK1/2 inhibition.

Example 12

BVD-523 in BRAF^V600EIn vivo antitumor Activity in mutant cancer cell line xenograft model

Based on our in vitro findings that BVD-523 reduces proliferation and induces apoptosis in a concentration-dependent manner, BVD-523 was administered by oral gavage to demonstrate its in vivo anti-tumor activity in a MAPK/ERK pathway-dependent model. Xenograft models using melanoma (cell line A375) and colorectal cancer (cell line Colo205), both carrying BRAF^V600EAnd (4) mutation.

BVD-523 efficacy was compared to the control cytotoxic alkylating agent temozolomide 14 days after treatment in a375 cell line xenografts. BVD-523 showed significant dose-dependent antitumor activity starting from 50mg/kg twice daily (BID) (fig. 31A). Doses of 50 and 100mg/kg BID significantly attenuated tumor growth with 71% (P ═ 0.004) and 99% (P <0.001), respectively, Tumor Growth Inhibition (TGI). 7 Partial Remissions (PR) were noted in the 100mg/kg BID group; no withdrawal response was found in any other group. The observed efficacy was superior to temozolomide, which when administered at 75 and 175mg/kg resulted in a moderate dose-dependent TGI of 34% (P >0.05) and 78% (P ═ 0.005), respectively.

In addition, BVD-523 showed anti-tumor efficacy in the Colo205 human colorectal cancer cell line xenograft model (FIG. 31B). BVD-523 again showed significant dose-dependent tumor regression at doses of 50, 75 and 100mg/kg BID, with mean tumor regression T/Ti (T ═ end of treatment, Ti ═ start of treatment) of-48.2%, -77.2% and-92.3%, respectively (all P < 0.0001). No regression was observed at the lowest dose of BVD-523(25mg/kg BID); however, significant tumor growth inhibition was observed with 25.2% T/C (T ═ treatment, C ═ control) (P < 0.0001). Although poorly tolerated, irinotecan (CPT-11), a positive control chemotherapeutic, showed significant antitumor activity with a T/C inhibiting Colo205 tumor growth of 6.4% (P < 0.0001). However, CPT-11 was not as effective as BVD-523 at 50, 75 or 100mg/kg BID doses, even at its maximum tolerated dose in mice.

To establish a relationship between pharmacokinetics and pharmacodynamics, BVD-523 plasma concentrations were compared to pERK1/2 levels measured in tumors by immunohistochemistry and isotopically labeled internal standard mass spectrometry over a 24 hour period following a single 100mg/kg oral dose of BVD-523 (fig. 31C). In untreated tumors, phosphorylation of ERK1/2 was low (0 hours). After treatment with BVD-523, ERK1/2 phosphorylation steadily increased from 1 hour post-dose to a maximum level of 8 hours post-dose, and then returned to pre-dose levels after 24 hours. This increase in pERK1/2 was correlated with BVD-523 drug plasma concentrations. The in vivo observation of the increase in pERK1/2 treated with BVD-523 was consistent with the earlier in vitro findings (FIG. 30D).

Example 13

Although ERK1/2 phosphorylation was increased, BVD-523 caused ERK1/2 substrate inhibition

To examine the effect of BVD-523 on signaling relative to other known ERK1/2 inhibitors (SCH772984, GDC-0994 and Vx-11e) (Morris et al, 2013 and Liu et al, 2015), a large scale reverse protein array (RPPA) of approximately 40 proteins was used for multiple cell lines sensitive to ERK inhibition. Cell lines with common alterations in BRAF and RAS were assayed: BRAF^V600EMutant system a375, Colo205 and HT 29; KRAS^G12C-mutant cell line MIAPACa-2; KRAS^G13D-the mutant cell line HCT 116; and with atypical HRAS^F82LMutant AN3 Ca. Changes in protein levels are shown as percent changes compared to the Dimethylsulfoxide (DMSO) -treated parental control (figure 32A and table 23). In addition to phosphorylation of ERK1/2 (pERK1/2[ ERK1/2-T202, -Y204)]) In addition, all ERK inhibitors elicit qualitatively similar protein effects; SCH7722984 inhibited pERK1/2 in all cell lines, while BVD-523, GDC-0994 and Vx-11e significantly increased pERK 1/2. Phosphate ion of the proximal and distal targets of pERK1/2, respectivelyp90RSK (pRSK1) and cyclin D1 were similarly inhibited by all inhibitors tested, regardless of the degree of ERK1/2 phosphorylation (fig. 32B). These independent findings for BVD-523 are consistent with studies showing that phosphorylation of the ERK1/2 substrate RSK1/2 is still inhibited, despite the significantly elevated pERK1/2 by Western blotting in A375 cells (FIG. 32D), in addition to protein binding studies demonstrating that BVD-523 binds and stabilizes pERK1/2 and inactive ERK1/2 (FIG. 29E and FIG. 29F). Thus, measurement of increased levels of pERK1/2 may be considered a clinical pharmacodynamic biomarker for BVD-523, while quantification of inhibition of ERK1/2 targets such as pRSK1 and DUSP6 may also play a similar role.

Additional protein changes were noted in the RPPA data set (fig. 32A). The reduced pS 6-ribosomal protein appeared to be another pharmacodynamic marker of ERK1/2 inhibition as demonstrated in all cell lines using all compounds (fig. 32B). Furthermore, significant induction of pAKT appeared to be a cell line dependent observation, with each ERK1/2 inhibitor inducing pAKT in cell lines a375 and AN3CA cells (fig. 33). Interestingly, the degree of inhibition of the survival marker pBAD appeared to vary between compounds, with GDC-0994 having only modest inhibition of pBAD compared to the other ERK1/2 inhibitors tested (fig. 32A).

Next, how BVD-523 affects BRAF was investigated^V600ECellular localization of ERK1/2 and downstream target pRSK in mutant RKO colorectal cell lines (FIG. 32C). In resting cells, ERK1/2 localizes to the cytoplasm and, upon stimulation, pERK1/2 migrates to target organelles, particularly the nucleus where the transcriptional target is activated (Wainstein et al, 2016). In DMSO-treated control cells, pERK1/2 was evident in both the nuclear and cytoplasmic fractions due to the presence of BRAF in this cell line^V600EThis may reflect MAPK pathway activity. Treatment with BVD-523 resulted in an increase in pERK1/2 in the nucleus and cytoplasm and a modest increase in total ERK1/2 in the nucleus compared to DMSO-treated cells, indicating that compound-induced pERK1/2 stabilization stimulated some nuclear translocation. Although pERK1/2 was increased in both compartments, pRSK levels were lower in the cytoplasmic and nuclear compartments compared to the DMSO control. Comparison MAPK SignalSignal transduction inhibitors (i.e., trametinib, SCH7722984, dabrafenib) inhibit phosphorylation of ERK1/2 and RSK as reflected by lower levels in the nuclear and cytoplasmic compartments. These data again indicate that the increase in BVD-523-associated pERK1/2 was evident in both cytoplasm and nucleus; however, this does not imply activation of the target substrate. This is consistent with the data presented in fig. 30D and fig. 32A.

Example 14

BVD-523 was shown to be active in an in vitro model of BRAF and MEK inhibitor resistance

The emergence of resistance to BRAF and MEK inhibitors limits their clinical efficacy. Here, experiments attempted to model and compare the development of resistance to BRAF (dabrafenib), MEK (trametinib) and ERK1/2(BVD-523) inhibition in vitro. In months, BRAF^V600EMutant a375 cells were cultured in increasing concentrations of each inhibitor. Drug-resistant a375 cell lines were readily obtained after growth in high concentrations of trametinib or dabrafenib, while development of a cell line resistant to BVD-523 proved challenging (fig. 34A). Taken together, these in vitro data indicate that delayed resistance to BVD-523 compared to dabrafenib or trametinib at concentrations that produce similar target inhibition may be interpreted as a durable response in the clinic.

Reactivation and dependence on ERK1/2 signaling is a common feature of acquired resistance to BRAF/MEK inhibition (Morris et al, 2013 and hatzivasssiliou et al, 2012); thus, the activity of BVD-523 in an in vitro model of acquired resistance was evaluated. First, the da rafenib and trametinib combination resistant a375 population was obtained using the concentration-increasing method described. The IC's of dabrafenib, trametinib and BVD-523 in the BRAF/MEK combination resistant population are shown in Table 24₅₀And IC compared to parent A375₅₀Fold change. BVD-523 IC₅₀Moderate shifts (2.5 fold), while dalafinib and trametinib shifted more significantly (8.5 and 13.5 fold, respectively) (table 24). Testing the cytotoxic agent paclitaxel asIn contrast, only modest changes in efficacy were observed. These data support the study of BVD-523 in the case of BRAF/MEK therapy resistance, although the resistance mechanism in this cell population remains to be characterized.

TABLE 24 BVD-523 Activity in a model of BRAF/MEK inhibition

To further investigate the ease of ERK1/2 inhibition in models with known BRAF inhibitor resistance mechanisms, AAV-mediated gene targeting was used to generate a pair of RKO BRAFs^V600EMutant cell line for MEK1^Q56PThe presence or absence of engineered heterozygous knockins that activate mutations is isogenic (Trunzer et al, 2013 and Emery et al, 2009). Including MEK1^Q56PThe MEK1/2 mutation within is associated with acquired resistance to single agent BRAF and combination BRAF/MEK therapies in patients (Wagle et al, 2011, Wagle et al, 2014, Emery et al, 2009 and Johnson et al, 2015). Single reagent assay demonstrated, relative to parental BRAF^V600E::MEK1^wtCell, double mutant BRAF^V600E::MEK1^Q56PCells showed significantly reduced sensitivity to the BRAF inhibitors vemurafenib and dabrafenib, and the MEK inhibitor trametinib (fig. 34B). In contrast, the response to BVD-523 was in the parent and MEK1^Q56PEssentially the same in the mutant cells, indicating that BVD-523 is not susceptible to this mechanism of acquired resistance. These results are in 2 independently derived double mutant BRAFs^V600E::MEK1^Q56POne was confirmed in cell line clones, thus validating the results with MEK1^Q56PThe presence of mutations was specifically correlated, but not unrelated cloning artifacts (data not shown). Similar results were also observed with a second, different inhibitor of ERK1/2 (SCH772984), supporting the expectation that these observations are specifically related to the mechanism of inhibition of ERK1/2, rather than due to off-target compound effects.

To further characterize BVD-523 pairs of BRAF^V600E::MEK1^Q56PMechanistic effects of MAPK pathway signaling in cell lines protein levels were assessed by Western blot (fig. 34C). In the parent BRAF^V600EIn RKO cells, a decrease in pRSK1/2 levels was observed after 4 hours of treatment with pharmacologically active concentrations of BRAF (Verofinib), MEK (trimetatinib) or ERK1/2(BVD-523) inhibitors. In contrast, isogenic double mutant BRAF^V600E::MEK1^Q56PCells did not show reduced RSK phosphorylation after treatment with BRAF or MEK inhibitors, while BVD-523 was still effective in inhibiting pRSK1/2 to levels comparable to parental RKO. Similarly, in the parents RKO and BRAF^V600E::MEK1^Q56PIn 24 hours BVD-523 treatment resulted in a decrease in pRB, indicating G0/G1 stasis.

Acquired KRAS mutations are also known drivers of resistance to MAPK pathway inhibitors. To understand the susceptibility of BVD-523 to this resistance mechanism, an isogenic panel of clinically relevant KRAS mutations in the colorectal cell line SW48 was used. The sensitivity to BVD-523 was compared to the MEK inhibitors semetitinib (selumetini) and trametinib (fig. 34D). Sensitivity to paclitaxel was unchanged (fig. 37A). Although several mutant KRAS alleles conferred robust to moderate levels of resistance to MEK inhibition, most alleles did not alter sensitivity to BVD-523, and when a shift in sensitivity was observed, it did not reach the extent observed with trametinib or semetinib. Taken together, these data indicate that BVD-523 is more effective than MEK inhibitors in this regard.

Example 15

BVD-523 shows in vivo activity in BRAF inhibitor resistant patient-derived melanoma xenograft model

To confirm and extend the anti-tumor effects of BVD-523 observed in an in vitro model of BRAF-/MEK-acquired resistance, a BRAF-resistant xenograft model derived from patients resistant to vemurafenib was used. BVD-523 was administered by oral gavage at 100mg/kg BID for 28 days, including alone and in combination with 50mg/kg BID of dabrafenib (FIG. 35). As expected, the single agent dabrafenib showed the lowest antitumor activity (22% TGI). Compared with a vehicle control, the BVD-523 activity is remarkable (P is less than or equal to 0.05), and the TGI is 78 percent. In this model, BVD-523 was combined with dabrafenib to give a TGI of 76% (P.ltoreq.0.05); thus, no further benefit was obtained in combination in this model of BRAF acquired resistance compared to the single agent BVD-523.

Example 16

Combination therapy with BVD-523 and BRAF inhibitors provides promising anti-tumor activity

Patients with BRAF mutant cancers may acquire resistance to combined BRAF/MEK therapy (Wagle et al, 2014), requiring consideration of other combination approaches within the MAPK pathway. In BRAF^V600EThe antiproliferative effect of the combination of BVD-523 with the BRAF inhibitor vemurafenib was evaluated in the mutant melanoma cell line G-361. As expected, both the single agents BVD-523 and vemurafenib were active, and when combined a modest synergistic effect was observed (fig. 37B). This indicates that BVD-523 in combination with a BRAF inhibitor carries BRAF^V600EThe mutated melanoma cell lines are at least additive and potentially synergistic. In addition, BRAF inhibitor plus BVD-523 was continuously cultured^V600EThe development of acquired resistance in vitro after mutant cell lines (a375) is challenging. In contrast, development of resistance to dabrafenib alone occurred relatively rapidly (fig. 37C). Even resistance to combined dabrafenib and trametinib occurred before dabrafenib plus trametinib.

The benefits of combined BRAF and ERK inhibition may not be fully realized in vitro combination studies where the concentration is not subject to tolerability limitations. To understand the benefits of this combination, a BRAF is used^V600EXenografts of mutant human melanoma cell line a375 efficacy was assessed in vivo. Due to the remarkable response to combination treatment, dosing in combination groups was stopped on day 20 to monitor tumor regrowth, and at 42The day starts again (fig. 36A). Tumors were measured twice weekly until study termination on day 45. The median Time To Endpoint (TTE) for the control was 9.2 days and the maximum possible Tumor Growth Delay (TGD) for 35.8 days was defined as 100%. Temozolomide treatment resulted in a TGD of 1.3 days (4%) with no regression. 50 and 100mg/kg Dalafinil monotherapies produced TGD of 6.9 days (19%) and 19.3 days (54%), respectively, showing significant survival benefit (P)<0.001) and 100mg/kg of 1PR in groups. 100mg/kg BVD523 monotherapy resulted in 9.3 days of TGD (26%), showing significant survival benefit (P)<0.001) and 2 persistent complete responses. The combination of dabrafenib and BVD-523 each produced the largest possible 100% TGD, had a noticeable withdrawal response, and had statistically higher overall survival (P) than its corresponding monotherapy<0.001). The lowest dose combination produced a remarkable 7/15 tumor-free survivor (TFS), and the 3 higher dose combinations produced a total of 43/44TFS, consistent with curative or near-curative activity (figure 36B). In conclusion, the combination of dabrafenib with BVD-523 produced a greater amount of TFS and efficacy over the single agent.

Based on BVD-523 Galrafenib, has a particle size of about 75-144mm³In the A375 xenograft model of initial tumor volume, follow-up experiments were performed to determine the "late" A375 xenografts (mean tumor initial volume, 700-³) Efficacy of (1) (fig. 36C). The median TTE for the control was 6.2 days, and the maximum possible TGD was determined to be 53.8 days, which was defined as 100% TGD for the 60 day study. BVD-523100 mg/kg monotherapy yielded negligible TGD (0.7 days, 1%) and no significant survival difference from the control group (P)>0.05). The distribution of TTE and 2 PR indicates that there may be a fraction of responders to treatment with BVD-523 alone. Darafenib 50mg/kg monotherapy was effective, had a TGD of 46.2 days (86%) and had significant survival benefit (P) compared to the control group<0.001). This group had 5 PR and 5 CR, including 3 TFS, in 11 evaluable mice (fig. 36D). Two combinations of dabrafenib and BVD-523 produced maximum 100% TGD and significant survival benefit (P) compared to the control group<0.001). Although there was a difference in the resolving activity, each combination100% regression response was produced in evaluable mice. The 25-mg/kg dabrafenib and 50-mg/kg BVD-523 combination had 2 PR and 8 CR with 6/10TFS, while the 50-mg/kg dabrafenib and 100-mg/kg BVD-523 combination had 11/11TFS on day 60 (fig. 36D). Overall, these data support the antecedent line combination of BVD-523 with BRAF targeted therapy at BRAF^V600The rationale in mutant melanoma, and this may extend to other tumor types that carry this change.

Discussion of the related Art

BVD-523 is a potent, highly selective, reversible small molecule ATP competitive inhibitor of ERK1/2, active in both in vivo and in vitro cancer models. In vitro, BVD-523 showed potent inhibition of several human tumor cell lines, particularly those carrying activating mutations in the MAPK signaling pathway, consistent with its mechanism of action. BVD-523 causes changes in downstream target and effector proteins, including direct substrate (pRSK) inhibition of ERK1/2 and total DUSP6 protein levels. These findings are consistent with previous findings on other ERK1/2 inhibitors demonstrating effective suppression of pRSK by ERK1/2 inhibition (Morris et al, 2013 and hatzivasssiliou et al, 2012). Interestingly, BVD-523 treatment resulted in a significant increase in ERK1/2 phosphorylation in vitro and in vivo. Similar to our findings, an increase in pERK1/2 was reported with the ERK1/2 inhibitor Vx11 e; in contrast, pERK1/2 inhibition occurred with SCH772984 (Morris et al, 2013). Although differences in pERK1/2 levels were observed in the various ERK1/2 inhibitors tested, downstream effectors (i.e., pRSK1 and total DUSP6) were similarly inhibited. These findings indicate that quantification of an ERK1/2 target substrate, such as pRSK1, can serve as a reliable pharmacodynamic biomarker for BVD-523-mediated inhibition of ERK1/2 activity.

Although BRAF (dabrafenib, vemurafenib) and MEK (trametinib, cobitinib) inhibitors validate the MAPK pathway as a therapeutic target, particularly in the presence of BRAF^V600In mutated patients, the anti-tumor response is limited by the emergence of acquired resistance and subsequent disease progression. Resistance is attributed to up-regulation and activation of compensatory signalling molecules (Nazari)an et al, 2010, villainueva et al, 2010, Johannessen et al, 2010 and Wang et al, 2011), amplification of target genes (Corcoran et al, 2010) and activating mutations of pathway components (e.g., RAS, MEK) (Wagle et al, 2011, Emery et al, 2009 and Wang et al, 2011). Reactivation of the ERK1/2 pathway is a common consequence of acquired resistance mechanisms. When BRAF is introduced^V600EMEK in the mutant melanoma cell line A375^Q56PConferring resistance to inhibition by MEK and BRAF (Wagle et al, 2011). In contrast, BVD-523 is in engineered MEK^Q56PRetention of potent inhibitory activity in cell lines indicates that ERK1/2 inhibition is effective in the context of upstream activation changes that may occur in response to BRAF/MEK treatment. As further evidence of the role of BVD-523 in the context of acquired resistance, the efficacy of BVD-523 was evident in the xenograft model of tumor samples from patients whose disease progressed on vemurafenib; the BRAF inhibitor dabrafenib is not effective in this model. These data support the role of targeting ERK1/2 in the case of BRAF/MEK resistance and complement previously published findings (Morris et al, 2013 and hatzivasssiliou et al, 2012). To further characterize resistance to inhibitors of the MAPK pathway, the emergence of resistance to BVD-523 itself was investigated. Single agent treatment of cancer cells with BVD-523 was found to be durable and more challenging to develop resistance compared to other agents that target upstream MAPK signaling components (i.e., dabrafenib, trametinib). This may indicate that achieving resistance to ERK1/2 targeting agents is more difficult to achieve than achieving resistance to BRAF or MEK therapy, possibly due to the fact that BVD-523 preferentially targets the more conserved active configuration of the ATP binding site. However, in vitro studies with other ERK1/2 inhibitors have identified specific mutants in ERK1/2 that drive resistance (Jha et al, 2016 and Goetz et al, 2014); these specific mutations have not been identified in clinical samples from patients with recurrent ERK1/2 inhibitors.

The potential clinical benefit of BVD-523 on ERK1/2 inhibition outweighs the case of BRAF/MEK therapy resistant patients. Since ERK1/2 is a downstream major node in this MAPK pathway, its inhibition is dependent on M for tumor growthAPK signaling is attractive in many cancer cases. Approximately 30% of all cancers carry RAS mutations; thus, targeting downstream ERK1/2 with BVD-523 is a rational treatment for these cancers. Furthermore, the results of the Hayes et al study showed that prolonged inhibition of ERK1/2 in KRAS mutant pancreatic cancer was associated with senescence-like growth inhibition (Hayes et al, 2016). However, in the case of RAS mutations, a combinatorial approach may be required to maximize and permanently attenuate MAPK signaling. For example, MEK inhibition in KRAS mutant colorectal cancer cells results in an adaptive response to ErbB family activation that inhibits the response to MEK inhibition (Sun et al, 2014). Similar background-specific adaptive responses may occur following ERK1/2 inhibition with BVD-523. The optimal therapeutic combination of various genetic profiles and cancer histology is the subject of ongoing research. BRAF removal^V600And RAS mutations, other changes driving MAPK are emerging. For example, new RAF fusions and atypical non-V600 BRAF mutations that promote RAF dimerization activate the MAPK pathway (Yao et al, 2015). Has been shown to inhibit BRAF^V600BRAF inhibitors of mutant monomeric proteins such as vemurafenib and dabrafenib are inactive in atypical RAF alterations that drive MAPK signaling in a dimerization-dependent manner (Yao et al, 2015). However, targeting downstream ERK1/2 in these tumors with treatment with BVD-523 may be a new approach to address this unmet medical need.

In BRAF^V600Combined BRAF and MEK inhibition exemplifies how agents targeting different nodes of the same pathway can improve therapeutic response and duration in the case of mutant melanoma tumors. We have BRAF in human melanoma cell line A375^V600Combination studies in mutant xenografts provide support for combination therapy with BVD-523 and BRAF inhibitors. The combination exhibits superior benefits over single agent treatment, including results consistent with a curative response. The clinical efficacy and tolerability of the combined BRAF/BVD-523 therapy remains to be determined. It is not unreasonable to expect that the BRAF/ERK1/2 combination is at least as functionally equivalent to the targeted BRAF/MEK combination. In addition, in comparison with other MAPK pathway inhibitorsThe more challenging in vitro observations of acquired resistance of BVD-523 suggest that BRAF/BVD-523 inhibitor combinations have the potential to provide a more durable response.

Significant progress has also been made using immunotherapy against melanoma. Various immune checkpoint inhibitors have been approved by the FDA in the united states for the treatment of advanced melanoma, including the cytotoxic T lymphocyte antigen-4 targeting agent ipilimumab (ipilimumab) and the programmed death-1 inhibitor pembrolizumab (pembrolizumab) and nivolumab (nivolumab). Combining BVD-523 with these immunotherapies is an attractive treatment option; further investigation is needed to explore the dosing schedule and assess whether a synergistic response can be achieved.

Based on preclinical data, BVD-523 may be expected to be useful for treating patients with malignancies that rely on MAPK signaling, including those whose tumors have acquired resistance to other treatments. The clinical development of BVD-523 is described below. See examples 17-24

Example 17

Homogeneous first-choice phase I dose escalation study of novel oral ERK1/2 kinase inhibitor BVD-523(ulixertinib) in patients with advanced solid tumors

The present invention describes a first dose escalation study of an ERK1/2 inhibitor in humans for the treatment of patients with advanced solid tumors. BVD-523 has an acceptable safety profile with good pharmacokinetics and early evidence of clinical activity.

Mitogen-activated protein kinase (MAPK) signaling through the RAS-RAF-MEK-ERK cascade plays a key role in tumorigenesis; and therefore have attracted considerable interest as therapeutic targets. This ubiquitous pathway consists of the cascade of the protein kinases RAF, MEK1/2 and ERK1/2, the upstream RAS. RAS is activated by GTP binding, which in turn leads to sequential activation of each protein kinase. While they appear to be the only physiological substrate for MEK1/2, ERK1/2 has many targets in the cytoplasm and nucleus, including the transcription factors Elk1, c-Fos, p53, Ets1/2, and c-Jun (Shaul et al, 2007). ERK1/2 activation and kinase activity affect cell proliferation, differentiation and survival through a variety of mechanisms (Rasola et al, 2010), including the activation of Ribosomal S6 Kinase (RSK) family members (Romeo et al, 2012).

Constitutive aberrant activation of the RAS-RAF-MEK1/2-ERK1/2 signaling pathway has been identified and is involved in the development or maintenance of many cancers (Schubbert et al, 2007 and Gollob et al, 2006). Mutations in RAS family genes (such as KRAS, NRAS and HRAS) are most common, with activating RAS mutations occurring in about 30% of human cancers (Schubbert et al, 2007). KRAS mutations are ubiquitous in: pancreatic cancer (> 90%) (Kanda et al, 2012), biliary cancer (3% -50%) (Hezel et al, 2014), colorectal cancer (30% -50%) (Arrington et al, 2012), lung cancer (27%) (pennyuick et al, 2012), ovarian cancer (15% -39%) (Dobrzycka et al, 2009), and endometrioid endometrial cancer (18%) (O' Hara and Bell, 2012); NRAS mutations are prevalent in melanoma (20%) (Khattak et al, 2013) and myeloid leukemia (8% -13%) (Yohe 2015); and HRAS mutations are common in bladder cancer (12%) (Fern ndez-Medarde and Santos, 2011). Mutations in RAF family genes (most particularly BRAF) occur frequently, particularly in melanoma. BRAF mutations have been found in 66% of malignant melanomas and-7% of other cancers (Davies et al, 2002), whereas MEK mutations are rare, with an overall frequency of 8% occurring in melanomas (Nikolaev et al, 2012). In contrast, few ERK mutations have been reported to date that lead to tumorigenesis (Deschenes-Simard et al, 2014).

The U.S. Food and Drug Administration (FDA) has approved two selective BRAF inhibitors, namely, vilotinib and daratinib, as BRAF^V600Monotherapy of mutant metastatic melanoma patients (Taflinar [ packaging insert ]]And Zelboraf [ packaging description]). Although the response rate of these targeted therapies is with BRAF^V600Up to 50% in patients with mutations, but the duration of response is usually measured in months rather than years (Hauschild et al, 2012 and McArthur et al),2014). The MEK1/2 inhibitor trametinib was also approved as a monotherapy in this case (Mekinist [ package insert ]]) But more commonly used in combination with the BRAF inhibitor, dalatinib. First-line use of trametinib administered in combination with dabertinib provided greater improvement in overall survival without increasing overall toxicity compared to vilotinib monotherapy (Robert et al, 2015), highlighting the potential utility of multiple proteins simultaneously targeting this MAPK signaling pathway. This therapeutic combination is also associated with MEK inhibitor-associated rash and a lower incidence of BRAF inhibitor-induced hyperproliferative skin lesions than each drug alone (Flaherty et al, 2012). Recently, phase III trials have also been shown in BRAF^V600E/KOverall survival of dabigatran alone in patients with mutation-positive melanoma (25.1 vs. 18.7 months, risk ratio [ HR ] with dabigatran etexilate]0.71, P ═ 0.0107), Progression Free Survival (PFS) (11.0 vs 8.8 months, HR 0.67, P ═ 0.0107), and overall response (69% vs 53%; p ═ 0.0014) (Long et al, 2015). Similarly, the combination of cobicistinib plus erlotinib demonstrated PFS (9.9 vs 6.2 months, HR 0.51, P) compared to vilotinib alone<0.001) and complete response rate (CR) or partial response rate (PR) (68% vs 45%; p<0.001) was significantly improved (Larkin et al, 2014). For this reason, the FDA recently approved a combination of vilotinib and cobitinib for BRAF^V600E/KA mutant melanoma. Based on these and related findings, the combination of a BRAF inhibitor plus a MEK inhibitor has become BRAF containing^V600E/KStandard targeted therapy selection of patients with mutated metastatic melanoma.

Although BRAF/MEK targeted combination therapy has been demonstrated to provide significant additional benefits over single agent selection, most patients eventually develop resistance and disease progression after about 12 months (Robert et al, 2015, Flaherty et al, 2012 and Long et al, 2015). Several mechanisms of acquired resistance have been identified following single agent or combination therapy, including the generation of BRAF splice variants, BRAF amplification, development of NRAS or MEK mutations, and upregulation of alternative pathways (pourikaos et al, 2011, Corcoran et al, 2010, Nazarian et al, 2010, Shi et al, 2014, Johannessen et al, 2010, Wagle et al, 2011, Wagle et al, 2014 and Ahronian et al, 2015). Central to many of these resistance mechanisms is reactivation of ERK signaling, which can rapidly restore MAPK pathway signaling and tumor cell escape from single agent BRAF or combination BRAF/MEK inhibitor therapy (pariiso et al, 2010). ERK inhibition may provide an opportunity to avoid or overcome resistance from upstream mechanisms, as it is the most distant master kinase of the MAPK signaling pathway. This is supported by preclinical evidence that inhibition of ERK by small molecule inhibitors both inhibits the emergence of resistance and overcomes acquired resistance to BRAF and MEK inhibitors (Morris et al, 2013 and hatzivasssiliou et al, 2012).

BVD-523 is a highly potent, selective, reversible ATP-competitive ERK1/2 inhibitor that has been shown to reduce tumor growth and induce tumor regression in BRAF and RAS mutant xenograft models. In addition, the single agent BVD-523 inhibits the human xenograft model with cross-resistance to both BRAF and MEK inhibitors. See examples 9-16. Thus, an open label, first human study (clinicaltrals. gov identifier, NCT01781429) of oral BVD-523 was performed to determine the maximum tolerated dose and the recommended dose for further studies. The study also aims to assess the pharmacokinetic and pharmacodynamic properties as well as the primary efficacy of patients with advanced cancer.

Example 18

Patient characteristics

A total of 27 patients enrolled and received at least one dose of study medication from 4/4 days 2013 to 12/1 days 2015. Baseline demographics and disease characteristics are shown in table 25. Median patients were 61 years of age (range, 33-86 years). Fifty-two percent (14/27) of the patients were male, and 63% (17/27) of the patients had an Eastern Cooperative Oncology Group (ECOG) status score of 1. Melanoma is the most common cancer (30%; BRAF mutation is present in 7/8 of these patients). The remaining patients had colorectal cancer (19%; 5/27), papillary thyroid cancer (15%; 4/27) or non-small cell lung cancer (NSCLC) (7%; 2/27), and 8 cases (30%) were classified as having other cancers (2 pancreas, 1 appendix, 1 non-seminal germ cell, 1 ovary, and 3 unknown primary). Most patients received 2 or more lines of prior systemic treatment, with 41% (11/27) receiving 2 to 3 lines and 48% (13/27) receiving >3 lines of prior systemic treatment.

TABLE 25 Baseline demographic and clinical characteristics of patients

^a7 are BRAF mutants, 1 is unknown.

^bTwo pancreases, 1 appendix, 1 non-spermatogonial germ cell, 1 ovary, 3 unknown primary.

^cPatients may have more than 1 molecular abnormality.

^dOther molecular abnormalities include ERCC1, RRM1, thymidylate synthase, GNAS, MEK1, TP53, CREBBP, ROS1, PTEN, AKT3, and PIK3 CA.

^eSome patients receive treatment with more than one BRAF inhibitor.

Abbreviations: ECOG, Eastern Cooperative Oncology Group (Eastern Cooperative Oncology Group).

Example 19

Ex vivo Effect of BVD-523 on RSK1/2 phosphorylation

Ex vivo useful for supporting clinical studies was developedBiomarker assay to demonstrate inhibition of ERK activity by BVD-523. This assay expands preclinical cellular data in which inhibitors of MAPK signaling, such as BVD-523, dabrafenib, trametinib, and verotinib, have been shown to inhibit RSK phosphorylation as a function of inhibitor concentration in BRAF mutant cancer cell lines. See examples 9-16. In particular, ERK inhibitor-dependent inhibition of phorbol 12-myristate 13-acetate (PMA) stimulated phosphorylation of the ERK substrate RSK1 in whole blood was used as target marker. PMA-stimulated RSK phosphorylation decreased with increasing BVD-523 concentration when BVD-523 was added directly to whole blood from healthy volunteers (FIG. 38). Average IC of accumulated data of BVD-523₅₀461. + -.20 nM, with a maximum inhibition of 75.8. + -. 2.7% at 10. mu.M BVD-523. Maximum inhibition was defined as RSK phosphorylation measured in the presence of 10. mu.M BVD-523. Patient-derived whole blood samples collected immediately prior to dosing or at defined time points after dosing with BVD-523 were similarly processed and RSK phosphorylation levels quantified.

Example 20

Dose escalation, Dose Limiting Toxicity (DLT), Maximum Tolerated Dose (MTD) and recommended phase II dose (RP2D)

According to the protocol, 5 single patient cohorts (10 to 150mg [ BID ] twice daily) progressed without signs of DLT. The 300mg BID cohort was expanded to more fully characterize the BVD-523 exposure. One of 6 patients given 600mg BID experienced grade 3 rash in DLT. At 900mg BID dose above MTD, one patient experienced grade 3 itching and elevated aspartate Aminotransferase (AST), and another patient experienced grade 3 diarrhea, vomiting, dehydration and elevated creatinine (table 26). The subsequent intermediate dose of 750mg BID also exceeded MTD, with 1 patient having grade 3 rash and grade 2 diarrhea in DLT, and another 1 patient having grade 2 hypotension, elevated creatinine and anemia. Therefore, MTD and RP2D were determined to be 600mg BID.

TABLE 26 dose limiting toxicity for round 1 (day 21)

^aIntermediate doses.

Abbreviations: AST, aspartate aminotransferase, BID, twice daily; DLT, dose-limiting toxicity; N/A, not applicable.

Example 21

Adverse Event (AE)

Treatment-related AEs assessed by any grade of investigator were noted in 26 of 27 patients (96%). The most common treatment-related AEs (> 30%) were skin rash (mainly acne) (70%), fatigue (59%), diarrhea (52%) and nausea (52%) (table 27). No patient experienced a grade 4 or 5 treatment-related AE or stopped treatment due to a treatment-related AE. Most events were on a grade 1 to 2, with 13 (48%) of 27 patients noticing treatment-related grade 3 events. The only grade 3 treatment-related event present in > 10% of patients was diarrhea (15%) and increased liver function check (11%), all of which occurred above the 600-mg BID dose.

TABLE 27-10% adverse events in patients that may/definitely correlate with BVD-523

^aNone of the patients experienced a grade 4 or 5 AE that may be or was clearly associated with BVD-523 treatment.

^bAxilla and maculopapular eruptions.

^cOne example of central serous retinopathy of relevanceA level 1 event.

Analysis expiration date: 12 months and 1 day 2015.

Abbreviations: AE, adverse event; ALT, alanine aminotransferase; AST, aspartate aminotransferase; LFT, liver function test.

A total of 28 severe aes (sae) were experienced by 14 patients. Of these 9 were considered by the investigator to be associated or likely associated with BVD-523 including dehydration, diarrhea or elevated creatinine (2 patients per case), emesis, nausea and fever (1 patient per case). All other SAEs are considered unrelated to treatment with BVD-523. Dose reduction by AE occurred in 3 patients during the study: 1 patient reduced from 600mg BID to 300mg BID, and 2 patients reduced from 900mg BID to 600mg BID.

Example 22

Pharmacokinetics

The single dose and steady state pharmacokinetics of BVD-523 are summarized in FIG. 39A and Table 28. In general, orally administered BVD-523 is slowly absorbed in patients with advanced malignancies. At the maximum concentration (C)_max) Thereafter, plasma BVD-523 levels were maintained for about 2 to 4 hours. Subsequently, the plasma drug concentration slowly decreased. Since plasma drug concentrations were only measured up to 12 hours after the morning dose, it was not possible to calculate the effective or terminal elimination rate. When administered up to 600mg BID, BVD-523 pharmacokinetics are linear and are C-specific_maxAnd area under the curve (AUC) is dose proportional. When the dose was increased from 600mg to 900mg BID, no further increase in exposure was observed. For all doses above 20mg BID, based on ex vivo whole blood assay C_maxTo EC₅₀Level (. apprxeq.200 ng/mL). Furthermore, steady-state exposure remained at the target EC for dose levels ≧ 150mg BID throughout the dosing period₅₀Or above the target EC₅₀. On day 15, at lower: (<75mg BID) dose level minimal plasma accumulation of BVD-523 and its metabolites was observed, whereas higher doses wereAccumulation at levels ranged from about 1.3-fold to 4.0-fold. The pre-dose concentrations at day 22 were generally similar to those at day 15, indicating that steady state had been reached at day 15 (data not shown). The degree of inter-patient variation of plasma exposure to BVD-523 and its metabolites was considered moderate and without problems.

TABLE 28 Steady-State BVD-523 pharmacokinetics (round 1, day 15)

^aDose levels administered twice daily;^bon day 15 n-3;^cthe number of subjects on day 15 at the 600mg dose level included two subjects who started on day 1 with 900mg dosing and then dropped to 600 mg;^don day 15 n-4;^fone subject started on day one with 750mg and finally dropped to 450 mg; day 15 parameters used in this study reflected at least 10 consecutive doses at 450 mg/dose. Individual day 15 parameters are 1300ng/mL C_maxAnd AUC of 10700_0-12；^gn＝3；^hn＝5。

At all dose levels over 12 hours post-dose, urinary excretion after the first dose of BVD-523 and at steady state was negligible (< 0.2% of dose) and was not dose-related in this very low percentage range. Renal clearance appears to be dose independent. Individual renal clearance values ranged from 0.128 to 0.0895L/hr (where n ≧ 1/dose level), with mean values ranging from 0.0149 to 0.0300L/hr (where n ≧ 3).

Example 23

Pharmacodynamic confirmation of targeted inhibition of BVD-523

To confirm targeting and pathway inhibition of BVD-523, RSK-1 phosphorylation was examined as a target biomarker in human whole blood samples from solid tumor patients receiving BVD-523. Steady-state whole blood samples collected from BVD-523 treated patients prior to day 15 dosing showed concentration-dependent inhibition of PMA-stimulated ERK activity (FIG. 39B), ranging from 0% ERK inhibition for BVD-523 dosed at 10mg BID to 93. + -. 8% ERK inhibition for 900mg BID dosed. The plasma concentrations of BVD-523 that gave 50% inhibition of ERK phosphorylation were similar whether BVD-523 was incorporated directly into the plasma of healthy volunteers or was present after oral administration to the patients.

Example 24

Anti-tumor effect

Tumor response to BVD-523 was evaluated in 25 evaluable patients using response evaluation criteria in Solid Tumors version 1.1 (RECIST v 1.1); 2 patients did not receive two scans of the target lesion and were therefore not evaluated using recistv 1.1. None of the patients achieved complete response, but 3 patients (all patients had BRAF-containing patients)^V600Mutant melanoma) all achieved partial response (129 days [ without BRAF/MEK-inhibitor treatment)]294 days in progress [ refractory to existing BRAF/MEK inhibitors]313 days in progress to date of data expiration [ intolerance to other BRAF/MEK inhibitors](FIG. 40A). Interestingly, all 3 partial responders had BRAF mutant melanoma. The total number of target lesions in one fraction of responders receiving BVD-523 at a 450mg BID dose was reduced by about 70% from baseline, while the other fraction of responders showed reductions of 47.0% and 33.6%. Stable disease was confirmed in 18 patients, 6 of which were stable for more than 6 months, and 6 others for more than 3 months. In this study, 4 patients exhibited progressive disease at the first evaluation.

Figure 40B shows a Computed Tomography (CT) scan of 1 out of 3 partial responders (RECIST v1.1) who progressed on previous verotinib and subsequent dalafinib/trametinib treatments; when BVD-523600 mg BID was administered>A permanent partial response was observed after 300 days. Of 5 of the 16 evaluable patients, BVD-523 was compared to positive emission tomography with fluorodeoxyglucose (X-ray tomography: (X-ray tomography) (II))¹⁸F-FDG-PET).

Figure 41 depicts response times and response durations in the study population. Two patients who responded to BVD-523 were still under study and continued BVD-523 treatment to the study expiration date (>500 days); in addition, one patient with bronchoalveolar NSCLC (tissue deficient for molecular profiling) had been treated for >700 days with stable disease. Of the 27 patients, 24 (90%) had discontinued treatment due to progressive disease (22/27, 82%) or other causes (2/27, 7%). The mean duration of BVD-523 treatment prior to discontinuation of treatment was 4.7 months.

Discussion of the related Art

The present invention provides the results of a first human study to evaluate BVD-523 in safety, pharmacokinetics, pharmacodynamics and primary efficacy in 27 patients with advanced solid tumors. In this dose escalation study, oral treatment of BVD-523 produced a radiographic response (3 partial responses) by RECIST v1.1 and prolonged disease stability in some patients, most of whom had received ≧ 2 prior systemic treatments. By pairs¹⁸Tumor uptake of F-glucose was imaged and evidence of BVD-523-dependent inhibition of metabolic response in tumors was established in a subset of patients. Drug exposure increased linearly with dose increase to 600mg BID, where exposure to 600mg BID provided nearly complete 24/7 inhibition of ERK-dependent substrate (RSK-1) phosphorylation in an ex vivo whole blood assay. Furthermore, when administered up to its MTD and RP2D (determined as 600mg BID), tolerance to BVD-523 was controlled.

BVD-523 is generally well tolerated with controlled and reversible toxicity. The most common AEs were skin rashes (usually acne), fatigue, and gastrointestinal side effects including nausea, vomiting, and diarrhea. The safety of BVD-523 is consistent with its selective inhibition of the MAPK pathway; the AE profile shows considerable overlap with MEK inhibitors. However, toxicity associated with any targeted therapy may include dependence on both the specific mechanism and the extent of target inhibition as well as any off-target effects (Zelboraf [ package insert ] and Hauschild et al, 2012). Ongoing and future studies will extend the efficacy and safety demonstrated in this dose escalation study and will guide how the unique features of the ERK inhibitor BVD-523 can be used as a single agent or in combination with other agents.

The sustained response of RAF and MEK inhibitors is often limited by intrinsic and ultimate acquired resistance, the common feature of which often involves reactivation of the ERK pathway (pourikos et al, 2011, Corcoran et al, 2010, Nazarian et al, 2010, Shi et al, 2014, Johannessen et al, 2010, Wagle et al, 2011, Wagle et al, 2014, Ahronian et al, 2015 and pariiso et al, 2010). Thus, inhibition of ERK by BVD-523 alone or in combination with other MAPK signaling pathway inhibitors has the potential to delay the development of resistance to existing therapies and to benefit a broader patient population. ERK inhibitors, including BVD-523, retain their efficacy in BRAF-and MEK-resistant cell lines, providing preclinical evidence for the use of ERK inhibitors in patients with acquired resistance to standard therapy (BRAF/MEK combination therapy). See, for example, examples 9-16. Importantly, in this study, patients with stable cancer progression after initial treatment with a BRAF inhibitor (vemurafenib) and subsequent treatment with a combination of BRAF and MEK inhibitors (dabrafenib/trametinib) had a partial response when receiving the single agent BVD-523. This patient was in the study for a total of 708 days by the study expiration date reported herein. Based in part on the anti-tumor effect observed in this patient, FDA has specified BVD-523 for the treatment of patients with unresectable or metastatic BRAF^V600A rapid pathway development program for patients with mutation-positive melanoma that is refractory to treatment with a BRAF and/or MEK inhibitor or has developed after treatment with a BRAF and/or MEK inhibitor. Accurately defining how BVD-523 best supports patient care (e.g., as a single agent or in various combinations) requires additional clinical studies.

In summary, this example provides data from evaluating BVD-523, a novel homogeneous first-time ERK inhibitor, as initial data for a dose escalation portion of a phase I study for treatment of advanced cancer patients. Continuous, twice daily oral treatment with BVD-523 produced an anti-tumor effect in several patients, including patients who had not received or progressed on existing MAPK pathway-targeted therapy. In this population of patients with advanced cancer, BVD-523 is generally well tolerated and toxicity controlled; MTD and RP2D were 600mg BID. BVD-523 exposure increased linearly to RP2D, and robust pharmacodynamic effects were evident at this dose level. An extension of this phase I clinical study is currently being conducted to confirm and extend the observations made during the dose escalation phase. In particular, patients are participating in an expanded cohort of molecular classifications across various tumor histologies (e.g., NRAS, BRAF, MEK, or ERK alterations). In addition, an expanded cohort is evaluating BVD-523 for use in cancer patients who have not received available MAPK pathway therapy, or who have progressed on such therapy.

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All documents cited in this application are incorporated herein by reference as if fully set forth herein.

Although illustrative embodiments of the present invention have been described herein, it is to be understood that the invention is not limited to those described, and that various other changes or modifications may be made by one skilled in the art without departing from the scope or spirit of the invention.

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agaccgatcc tcatcagctc ccaatgtgca tataaacaca atagaacctg tcaatattga 1200

tgacttgatt agagaccaag gatttcgtgg tgatggagga tcaaccacag gtttgtctgc 1260

taccccccct gcctcattac ctggctcact aactaacgtg aaagccttac agaaatctcc 1320

aggacctcag cgagaaagga agtcatcttc atcctcagaa gacaggaatc gaatgaaaac 1380

acttggtaga cgggactcga gtgatgattg ggagattcct gatgggcaga ttacagtggg 1440

acaaagaatt ggatctggat catttggaac agtctacaag ggaaagtggc atggtgatgt 1500

ggcagtgaaa atgttgaatg tgacagcacc tacacctcag cagttacaag ccttcaaaaa 1560

tgaagtagga gtactcagga aaacacgaca tgtgaatatc ctactcttca tgggctattc 1620

cacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagct tgtatcacca 1680

tctccatatc attgagacca aatttgagat gatcaaactt atagatattg cacgacagac 1740

tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc tcaagagtaa 1800

taatatattt cttcatgaag acctcacagt aaaaataggt gattttggtc tagctacagt 1860

gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca ttttgtggat 1920

ggcaccagaa gtcatcagaa tgcaagataa aaatccatac agctttcagt cagatgtata 1980

tgcatttgga attgttctgt atgaattgat gactggacag ttaccttatt caaacatcaa 2040

caacagggac cagataattt ttatggtggg acgaggatac ctgtctccag atctcagtaa 2100

ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc tcaaaaagaa 2160

aagagatgag agaccactct ttccccaaat tctcgcctct attgagctgc tggcccgctc 2220

attgccaaaa attcaccgca gtgcatcaga accctccttg aatcgggctg gtttccaaac 2280

agaggatttt agtctatatg cttgtgcttc tccaaaaaca cccatccagg cagggggata 2340

tggtgcgttt cctgtccact gaaacaaatg agtgagagag ttcaggagag tagcaacaaa 2400

aggaaaataa atgaacatat gtttgcttat atgttaaatt gaataaaata ctctcttttt 2460

ttttaaggtg aaccaaagaa cacttgtgtg gttaaagact agatataatt tttccccaaa 2520

ctaaaattta tacttaacat tggattttta acatccaagg gttaaaatac atagacattg 2580

ctaaaaattg gcagagcctc ttctagaggc tttactttct gttccgggtt tgtatcattc 2640

acttggttat tttaagtagt aaacttcagt ttctcatgca acttttgttg ccagctatca 2700

catgtccact agggactcca gaagaagacc ctacctatgc ctgtgtttgc aggtgagaag 2760

ttggcagtcg gttagcctgg gttagataag gcaaactgaa cagatctaat ttaggaagtc 2820

agtagaattt aataattcta ttattattct taataatttt tctataacta tttcttttta 2880

taacaatttg gaaaatgtgg atgtctttta tttccttgaa gcaataaact aagtttcttt 2940

ttataaaaa 2949

<210>2

<211>765

<212>PRT

<213> human

<400>2

Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Ala Glu Pro Gly Gln Ala

1 5 10 15

Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala Gly Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Met Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ala

130 135 140

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

275 280 285

Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala

290 295 300

Ser Leu Ala Glu Thr Ala Leu Thr Ser Gly Ser Ser Pro Ser Ala Pro

305 310 315 320

Ala Ser Asp Ser Ile Gly Pro Gln Ile Leu Thr Ser Pro Ser Pro Ser

325 330 335

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

340 345 350

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

355 360 365

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp

370 375 380

Gln Gly Phe Arg Gly Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr

385390 395 400

Pro Pro Ala Ser Leu Pro Gly Ser Leu Thr Asn Val Lys Ala Leu Gln

405 410 415

Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu

420 425 430

Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp

435 440 445

Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser

450 455 460

Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala

465 470 475 480

Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala

485 490 495

Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile

500 505 510

Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr

515 520 525

Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu

530 535 540

Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala

545 550 555 560

Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu

565 570 575

Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly

580 585 590

Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln

595 600 605

Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile

610 615 620

Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala

625 630 635 640

Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser

645 650 655

Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr

660 665 670

LeuSer Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met

675 680 685

Lys Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro

690 695 700

Leu Phe Pro Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu

705 710 715 720

Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala Gly

725 730 735

Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys Thr

740 745 750

Pro Ile Gln Ala Gly Gly Tyr Gly Ala Phe Pro Val His

755 760 765

<210>3

<211>3906

<212>DNA

<213> rat (Rattus norvegicus)

<400>3

atggcggcgc tgagtggcgg cggtggcagc agcagcggtg gcggtggcgg cggcggcggc 60

ggcggtggtg gcggcggcgg cggcggcgcc gaacagggac aggctctgtt caatggcgac 120

atggagccgg aggccggcgc tggcgccgcg gcctcttcgg ccgcggaccc ggccattcct 180

gaagaggtgt ggaatatcaa gcaaatgatt aagttgacac aggaacatat agaggcccta 240

ttggacaagt ttggtgggga gcataaccca ccgtcaatat acctggaggc ctatgaagag 300

tacaccagca agctagatgc ccttcagcag agagagcagc agctgttgga atccctggtt 360

tttcaaactc ccacagatgt atcacggaac aaccccaagt caccacagaa acctatcgtt 420

cgtgtcttcc tgcccaacaa acagaggaca gtggtgcccg caagatgtgg tgtaacggtc 480

cgagacagtc taaagaaagc actaatgatg aggggtctca tcccagagtg ctgtgctgtt 540

tacagaattc aggacggaga gaagaaacca attggctggg acactgacat ttcctggctt 600

actggagagg agctacatgt tgaagtacta gagaatgttc ctctgacaac ccacaacttc 660

gtacggaaaa cttttttcac cttagcattt tgtgactttt gccgaaagct gcttttccag 720

ggtttccgct gtcaaacatg tggttataag tttcaccagc gttgtagtac agaggttcca 780

ctgatgtgtg ttaattatga ccaacttgat ttgctgtttg tctccaagtt ctttgagcat 840

cacccagtac cacaggagga ggccttctca gcagagacta cccttccatc tggatgctct 900

tccgcacccc cctcagactc tattgggccc caaatcctca ccagtccatc tccttcaaaa 960

tccattccaa ttccacagcc cttccggcca gcagatgaag atcatcgcaa tcagtttggg 1020

caacgagacc gctcctcctc cgctcccaat gttcatataa acacaatcga acctgtcaat 1080

attgatgaaa aattcccaga agtggaatta caggatcaaa gggatttgat tagagaccag 1140

gggtttcgtg gggatggagc ccctttgaac cagctgatgc gctgtcttcg gaaataccaa 1200

tcccggactc ccagccccct cctccattct gtccccagtg aaatagtgtt tgattttgag 1260

cctggcccag tgttcagagg gtcaaccaca ggcttgtcgg ccaccccacc tgcctcatta 1320

cctggctcac tcactaacgt gaaagcctta cagaaatctc caggacctca gcgggaaagg 1380

aagtcctcct cctcctcctc ctccacggaa gacagaagtc ggatgaaaac acttggtaga 1440

agagattcaa gtgatgattg ggagattcct gatggacaga ttacagtggg acagagaatt 1500

ggatctgggt cctttggaac tgtctacaag ggaaagtggc atggcgacgt ggcagtgaaa 1560

atgctgaatg tgacagcacc cacacctcag cagttacagg ccttcaaaaa cgaagtcgga 1620

gtactcagga aaactcgaca tgtgaacatc ctccttttca tgggctattc tacaaagcca 1680

cagctggcta ttgttacaca gtggtgtgaa ggctccagct tatatcacca tctccacatc 1740

attgagacca aatttgagat gatcaaactt atagatattg cacggcagac tgcacagggc 1800

atggattact tacacgccaa gtcaatcatc cacagagacc tcaagagtaa taatatattt 1860

cttcatgaag acctcacggt aaaaataggt gactttggtt tagccacagt gaagtcccga 1920

tggagtgggt cccatcagtt tgaacagttg tctggatcta ttttgtggat ggcacccgaa 1980

gtaatcagaa tgcaagataa aaacccatat agctttcagt cagacgtgta tgcatttggg 2040

attgttctgt atgaactgat gactggtcag ctaccttatt caaacatcaa caacagggat 2100

cagataattt ttatggtggg acgaggatac ctatctccag atctcagtaa ggtacggagt 2160

aactgtccaa aagccatgaa gagattaatg gcagagtgcc tcaaaaagaa aagagacgag 2220

agaccactct ttccccaaat tctcgcctct attgagctgc tggcccgctc attgccaaaa 2280

attcaccgca gtgcatcaga accctccttg aatcgggctg gtttccaaac agaagatttt 2340

agtctgtatg cttgtgcttc tccaaaaaca cccatccaag cagggggata tggagaattt 2400

gcagccttca agtagccact ccatcatggc agcatctact ctttatttct taagtcttgt 2460

gttcatacaa tttgttaaca tcaaaacaca gttctgttcc tcaaattttt tttaaagata 2520

caaaattttc aatgcataag ctcgtgtgga acagaatgga atttcctatt caacaaaaga 2580

gggaagaatg ttttaggaac cagaattctc tgctgcccgt gtttcttctt caacacaaat 2640

atcatgtgca tacaactctg cccattccca agaagaaaga ggagagaccc cgaattctgc 2700

ccttttggtg gtcaggcatg atggaaagaa tttgctgctg cagcttggga aaaattgcta 2760

tggaaagtct gccagtcaac tttgcccttc taaccaccag atccatttgt ggctggtcat 2820

ctgatggggc gatttcaatc accaagcatc gttcttgcct gttgtgggat tatgtcgtgg 2880

agcactttcc ctatccacca ccgttaattt ccgagggatg gagtaaatgc agcataccct 2940

ttgtgtagca cctgtccagt cctcaaccaa tgctatcaca gtgaagctct ttaaatttaa 3000

gtggtgggtg agtgttgagg agagactgcc ttgggggcag agaaaagggg atgctgcatc 3060

ttcttcctca cctccagctc tctcacctcg ggttgccttg cacactgggc tccgcctaac 3120

cactcgggct gggcagtgct ggcacacatt gccgcctttt ctcattgggt ccagcaattg 3180

agcagagggt tgggggattg tttcctccac aatgtagcaa attctcagga aaatacagtc 3240

catatcttcc tctcagctct tccagtcacc aaatacttac gtggctcctt tgtccaggac 3300

ataaaacacc gtggacaaca cctaattaaa agcctacaaa actgcttact gacagttttg 3360

aatgtgagac atttgtgtaa tttaaatgta aggtacaggt cttaatttct tctattaagt 3420

ttcttctatt tttatttaaa cgaagaaaat aattttcagg tttaattgga ataaacgaat 3480

acttcccaaa agactatata ccctgaaaat tatatttttg ttaattgtaa acaactttta 3540

aaaaatggtt attatccttt tctctaccta aaattatggg aaatcttagc ataatgacaa 3600

ttatttatac tttttaaata aatggtactt gctggatcca cactaacatc tttgctaaca 3660

ttcccattgt ttcttccaac ttcactccta cactacatcc tccatcctct ttctagtctt 3720

ttatctataa tatgcaacct aaaataaaag tggtggtgtc tccattcatt cttcttcttc 3780

cttttttccc caagcctggt cttcaaaagg ttgggtaatt tagtagctga gttccctagg 3840

tagaaataga actattaggg acattggggt tgtaggaaag cgtgaggcct gtcaccagtt 3900

gttctt 3906

<210>4

<211>804

<212>PRT

<213> rat

<400>4

Met Ala Ala Leu Ser Gly Gly Gly Gly Ser Ser Ser Gly Gly Gly Gly

1 5 10 15

Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu Gln

20 25 30

Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala Gly

35 40 45

Ala Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp

50 55 60

Asn Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu

65 70 75 80

Leu Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu

85 90 95

Ala Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu

100 105 110

Gln Gln Leu Leu Glu Ser Leu Val Phe Gln Thr Pro Thr Asp Val Ser

115120 125

Arg Asn Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

130 135 140

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

145 150 155 160

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

165 170 175

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

180 185 190

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

195 200 205

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

210 215 220

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

225 230 235 240

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

245 250 255

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

260 265 270

Phe Val Ser Lys Phe Phe Glu His His Pro Val Pro Gln Glu Glu Ala

275 280 285

Phe Ser Ala Glu Thr Thr Leu Pro Ser Gly Cys Ser Ser Ala Pro Pro

290 295 300

Ser Asp Ser Ile Gly Pro Gln Ile Leu Thr Ser Pro Ser Pro Ser Lys

305 310 315 320

Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His Arg

325 330 335

Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val His

340 345 350

Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Glu Lys Phe Pro Glu Val

355 360 365

Glu Leu Gln Asp Gln Arg Asp Leu Ile Arg Asp Gln Gly Phe Arg Gly

370 375 380

Asp Gly Ala Pro Leu Asn Gln Leu Met Arg Cys Leu Arg Lys Tyr Gln

385 390 395 400

Ser Arg Thr Pro Ser Pro Leu LeuHis Ser Val Pro Ser Glu Ile Val

405 410 415

Phe Asp Phe Glu Pro Gly Pro Val Phe Arg Gly Ser Thr Thr Gly Leu

420 425 430

Ser Ala Thr Pro Pro Ala Ser Leu Pro Gly Ser Leu Thr Asn Val Lys

435 440 445

Ala Leu Gln Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser

450 455 460

Ser Ser Ser Ser Thr Glu Asp Arg Ser Arg Met Lys Thr Leu Gly Arg

465 470 475 480

Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly Gln Ile Thr Val

485 490 495

Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys

500 505 510

Trp His Gly Asp Val Ala Val Lys Met Leu Asn Val Thr Ala Pro Thr

515 520 525

Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val Gly Val Leu Arg Lys

530 535 540

Thr Arg His Val Asn Ile Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro

545 550 555 560

Gln Leu Ala Ile Val Thr Gln Trp Cys Glu Gly Ser Ser Leu Tyr His

565 570 575

His Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys Leu Ile Asp

580 585 590

Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His Ala Lys Ser

595 600 605

Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe Leu His Glu Asp

610 615 620

Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr Val Lys Ser Arg

625 630 635 640

Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp

645 650 655

Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe

660 665 670

Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu Tyr Glu Leu Met Thr

675680 685

Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe

690 695 700

Met Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys Val Arg Ser

705 710 715 720

Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys Leu Lys Lys

725 730 735

Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Ile Leu Ala Ser Ile Glu

740 745 750

Leu Leu Ala Arg Ser Leu Pro Lys Ile His Arg Ser Ala Ser Glu Pro

755 760 765

Ser Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala

770 775 780

Cys Ala Ser Pro Lys Thr Pro Ile Gln Ala Gly Gly Tyr Gly Glu Phe

785 790 795 800

Ala Ala Phe Lys

<210>5

<211>9728

<212>DNA

<213> mouse (Mus musculus)

<400>5

ccctcaggct cggctgcgcc ggggccgccg gcgggttcca gaggtggcct ccgccccggc 60

cgctccgccc acgccccccg cgcctccgcg cccgcctccg cccgccctgc gcctcccttc 120

cccctccccg ccccgcggcg gccgctcggc ccggctcgcg cttcgaagat ggcggcgctg 180

agtggcggcg gtggcagcag cagcggtggc ggcggcggcg gtggcggcgg cggtggcggt 240

ggcgacggcg gcggcggcgc cgagcagggc caggctctgt tcaatggcga catggagccg 300

gaggccggcg ctggcgccgc ggcctcttcg gctgcggacc cggccattcc tgaagaggta 360

tggaatatca agcaaatgat taagttgaca caggaacata tagaggccct attggacaaa 420

tttggtggag agcataaccc accatcaata tacctggagg cctatgaaga gtacaccagc 480

aagctagatg cccttcagca aagagaacag cagcttttgg aatccctggt ttttcaaact 540

cccacagatg catcacggaa caaccccaag tcaccacaga aacctatcgt tagagtcttc 600

ctgcccaaca aacagaggac agtggtaccc gcaagatgtg gtgttacagt tcgagacagt 660

ctaaagaaag cactgatgat gagaggtctc atcccagaat gctgtgctgt ttacagaatt 720

caggatggag agaagaaacc aattggctgg gacacggaca tttcctggct tactggagag 780

gagttacatg ttgaagtact ggagaatgtc ccacttacaa cacacaactt tgtacggaaa 840

acttttttca ccttagcatt ttgtgacttt tgccgaaagc tgcttttcca gggtttccgt 900

tgtcaaacat gtggttataa atttcaccag cgttgtagta cagaggttcc actgatgtgt 960

gtaaattatg accaacttga tttgctgttt gtctccaagt tctttgagca tcacccagta 1020

ccacaggagg aggcctcctt cccagagact gcccttccat ctggatcctc ttccgcaccc 1080

ccctcagact ctactgggcc ccaaatcctc accagtccat ctccttcaaa atccattcca 1140

attccacagc ccttccgacc agcagatgaa gatcatcgca atcagtttgg gcaacgagac 1200

cggtcctcct cagctcccaa tgttcatata aacacaattg agcctgtgaa tatcgatgaa 1260

aaattcccag aagtggaatt acaggatcaa agggatttga ttagagacca ggggtttcgt 1320

ggtgatggag cccccttgaa ccaactgatg cgctgtcttc ggaaatacca atcccggact 1380

cccagccccc tcctccattc tgtccccagt gaaatagtgt ttgattttga gcctggccca 1440

gtgttcagag ggtcaaccac aggcttgtcc gccaccccgc ctgcctcatt acctggctca 1500

ctcactaacg tgaaagcctt acagaaatct ccaggtcctc agcgggaaag gaagtcatct 1560

tcttcctcat cctcggagga cagaagtcgg atgaaaacac ttggtagaag agattcaagt 1620

gatgactggg agattcctga tggacagatt acagtgggac agagaattgg atctgggtca 1680

tttggaactg tctacaaggg aaagtggcat ggtgatgtgg cagtgaaaat gttgaatgtg 1740

acagcaccca cacctcaaca gctacaggcc ttcaaaaatg aagtaggagt gctcaggaaa 1800

actcgacatg tgaatatcct ccttttcatg ggctattcta caaagccaca actggcaatt 1860

gttacacagt ggtgtgaggg ctccagctta tatcaccatc tccacatcat tgagaccaaa 1920

tttgagatga tcaaacttat agatattgct cggcagactg cacagggcat ggattactta 1980

cacgccaagt caatcatcca cagagacctc aagagtaata atatatttct tcatgaagac 2040

ctcacggtaa aaataggtga ctttggtcta gccacagtga aatctcggtg gagtgggtcc 2100

catcagtttg aacagttgtc tggatctatt ttgtggatgg caccagaagt aatcagaatg 2160

caagataaaa acccgtatag ctttcagtca gacgtgtatg cgtttgggat tgttctgtac 2220

gaactgatga ccggccagct accttattca aacatcaaca acagggatca gataattttt 2280

atggtgggac gaggatacct atctccagat ctcagtaagg tacggagtaa ctgtccaaaa 2340

gccatgaaga gattaatggc agagtgcctc aaaaagaaaa gagacgagag accactcttt 2400

ccccaaattc tcgcctccat tgagctgctg gcccgctcat tgccaaaaat tcaccgcagt 2460

gcatcagaac cttccttgaa tcgggctggt ttccaaacag aagattttag tctgtatgct 2520

tgtgcttctc cgaaaacacc catccaagca gggggatatg gagaatttgc agccttcaag 2580

tagccagtcc atcatggcag catctactct ttatttctta agtcttgtgt tcatacagtt 2640

tgttaacatc aaaacacagt tctgttcctc aaaaaatttt ttaaagatac aaaattttca 2700

atgcataagt tcatgtggaa cagaatggaa tttcctattc aacaaaagag ggaagaatgt 2760

tttaggaacc agaattctct gctgcccgtg tttcttcttc aacataacta tcacgtgcat 2820

acaagtctgc ccattcccaa gaagaaagag gagagaccct gaattctgcc cttttggtgg 2880

tcaggcatga tggaaagaat ttgctgctgc agcttgggaa aattgctatg gaaagtctgc 2940

cagtcgactt tgcccttcta accaccagat cagcctgtgg ctggtcatct gatggggcga 3000

tttccatcac caagcatcgt tcttgcctat tctgggatta tgttgtggag cactttccct 3060

gtccagcacc gttcatttct gagggatgga gtaaatgcag cattcccttg tgtagcgcct 3120

gttcagtcct cagcagctgc tgtcacagcg aagcttttta cagttaagtg gtgggggaga 3180

gttgaggaga gcctgcctcg gggcagagaa aagggggtgc tgcatcttct tcctcacctc 3240

cagctctctc acctcgggtt gccttgctca ctgggctccg cctaaccact caggctgctc 3300

agtgctggca cacattgcct tcttttctca ttgggtccag caattgagga gagggttggg 3360

ggattgtttc ctcctcaatg tagcaaattc tcaggaaaat acagtccata tcttcctctc 3420

agctcttcca gtcaccaaat acttacgtgg ctcctttgtc caggacataa aacaccgtgg 3480

acaacaccta attaaaagcc tacaaaactg cttactgaca gttttgaatg tgagacactt 3540

gtgtaattta aatgtaaggt acaggtttta atttctgagt ttcttctatt tttatttaaa 3600

agaagaaaat aattttcagt tttaattgga ataaatgagt acttcccaca agactatata 3660

ccctgaaaat tatatttttg ttaattgtaa acaactttta aagaataatt attatccttt 3720

tctctaccta aaaattatgg ggaatcttag cataatgaca attatttata ctttttaaat 3780

aaatggtact tgctggatcc acactaacat ctttgctaac aatcccattg tttcttccaa 3840

cttaactcct acactacatc ctacatcctc tttctagtct tttatctata atatgcaacc 3900

taaaataaac gtggtggcgt ctccattcat tctccctctt cctgttttcc ccaagcctgg 3960

tcttcaaaag gttgggtaat cggtccctga gctccctagc tggcaatgca actattaggg 4020

acattggagt tgcaggagag caggaagcct gtccccagct gttcttctag aaccctaaat 4080

cttatctttg cacagatcaa aagtatcacc tcgtcacagt tctccttagc ctttacttac 4140

aggtaatata aataaaaatc accatagtag taaagaaaac aactggatgg attgatgacc 4200

agtacctctc agagccagga atcttgaatc tccaggattt atacgtgcaa atttaaggag 4260

atgtacttag caacttcaag ccaagaactt ccaaaatact agcgaatcta aaataaaatg 4320

gaattttgag ttatttttaa agttcaaatt ataattgata ccactatgta tttaagccta 4380

ctcacagcaa gttagatgga ttttgctaaa ctcattgcca gactgtggtg gtggtggtgg 4440

tagtgtgcac ctttaatcca agcaactcag caatcagaat gaggtaaatc tctgtgaata 4500

caaggcctgc ctagtctgca gcgctagttc caggatagcc agggctacac acacaaaaac 4560

cctctctcaa aaaaaacaaa attaattagt tgataataaa aaataactaa agtatcatca 4620

aaggaaggcc tactggaagt tttatatatt cccagtaaat tgaaaaatat tctgaagtta 4680

ttaaccagtt agcaacaatg tgtttttaag tcttacataa acagagcaaa gtcttcaaat 4740

gtttcagagc tgagaagata attgtgcttg atatgaaaaa tagcctctcc atatgatgtg 4800

ccacattgaa aggcgtcatt acccttttaa atacttctta atgtggcttt gttcccttta 4860

cccaggatta gctagaaaga gctaggtagg cttcggccac agttgcacat ttcgggcctg 4920

ctgaagaatg ggagctttga aggctggcct tggtggagga gcccctcagt gctggagggt 4980

ggggcgtgta cgcagcatgg aagtggtcta gacagagtgc aaagggacag acttctttct 5040

cattttagta tagggtgatg tctcacttga aatgagaaag tagagttgat attaaacgaa 5100

gctgtgccca gaaaccaggc tcagggtatt gtgagatttt ctttttaaat agagaatata 5160

aaagatagaa ataaatattt aaaccttcct tcttattttc tatcaaatag atttttttta 5220

tcatttgcaa acaacataaa aaaaggtttc ttttgtgggg ttttctttcc ttcttttttt 5280

tttttttttt tttttaagac tgcagataat cttgttgagc tcctcggaaa atacaaggaa 5340

gtccgtgttt gtgcagagcg ctttatgagt aactgtatag acagtgtggc tgcttcactc 5400

atcccagagg gctgcagctg tcggcccatg aagtggctgc agtgcctcgt gagatctgct 5460

ttgttttgtt tggagtgaag tctttgaaag gtttgagtgc aactatatag gactgttttt 5520

aaataagtag tattcctcat gaactttctc attgttaagc tacaggaccc aaactctacc 5580

actaagatat tattaacctc aaaatgtagt ttatagaagg aatttgcaaa tagaatatcc 5640

agttcgtact tatatgcatc ttcaacaaag attctctgtg acttgttgga tttggttcct 5700

gaacagccca tttctgtatt tgaggttagg agggcataat gaggcatcct aaaagacaat 5760

ctgatataaa ctgtatgcta gatgtatgct ggtaggggag aaagcattct gtaaagacat 5820

gatttaagac ttcagctctg tcaaccagaa accttgtaaa tacttcctgt cttggtgcag 5880

ccccgcccct ttgatcacac gatgttgtct tgtgcttgtc agacactgtc agagctgctg 5940

ttcgtccctc tgcagatctc acctgtcccc actgcacacc cacctcctgc ctcttgcaga 6000

cctcagcatc tagctttagt tggaaacagt tcagggttca ggtgacttct taaaaaaaaa 6060

aaaaaaccct acctcctcag aatgaggtaa tgaatagtta tttatttaaa gtatgaagag 6120

tcaggagcgc tcgaacatga aggtgattta agatggttcc tttcgtgtgt attgtagctg 6180

agcacttgtt tttgtcctaa agggcattat acatttaagc agtgattctg tttaaagatg 6240

tttttcttta aaggtgtagc tcagagtatc tgttgttgga attggtgcca gagtctgctt 6300

aatagatttc agaatcctaa gcttaagtca gtcgcatgaa gttaagtagt tatggtaaca 6360

ctttgctagc catgatataa ttctactttt taggagtagg tttggcaaaa ctgtatgcct 6420

tcaaagtgag ttggccacag ctttgtcaca tgcacagata ctcatctgaa gagactgccc 6480

agctaagagg gcggaaggat accctttttt cctacgattc gcttctttgt ccacgttggc 6540

attgttagta ctagtttatc agcaccttga ccagcagatg tcaaccaata agctattttt 6600

aaaaccatag ccagagatgg agaggtcact gtgagtagaa acagcaggac gcttacagga 6660

gtgaaatggt gtagggaggc tctagaaaaa tatcttgaca atttgccaaa tgatcttact 6720

gtgccttcat gatgcaataa aaaagctaac attttagcag aaatcagtga tttacgaaga 6780

gagtggccag tctggtttaa ctcagctggg ataatatttt tagagtgcaa tttagactgc 6840

gaagataaat gcactaaaga gtttatagcc aattcacatt tgaaaaataa gaaaatggta 6900

aattttcagt gaaatatttt tttaaagcac ataatcccta gtgtagccag aaatatttac 6960

cacatagagc agctaggctg agatacagtc cagtgacatt tctagagaaa ccttttctac 7020

tcccacgggc tcctcaaagc atggaaattt tatacaaaat gtttgacatt ttaagatact 7080

gctgtagttt agttttgaaa tagtatgtgc tgagcagcaa tcatgtacta actcagagag 7140

agaaaacaac aacaaattgt gcatctgatt tgttttcaga gaaatgctgc caacttagat 7200

actgagttct cagagcttca agtgtaaact tgcctcccaa gtcctgtttg caaatgaagt 7260

tggctagtgc tactgactgc tccagcacat gatggaaggc agggggctgt ctctgaagtg 7320

tcttctataa agggacaata gaatagtgag agacctggtc agtgtgtgtc agctggacac 7380

tccatgctat gggacttgca tcttctgtcc tcaccatccc caagacattg tgctttcctc 7440

agttgtcctc tagctgtttc actcagacac caagatgaat tactgatgcc agaaggggcc 7500

aaaatggcca gtgtgttttg ggggttgtat cagttgactg gacaataact ttaatagttt 7560

cagatcattt atttttactt ccattttgac agacatttaa atggaaattt agtcctaact 7620

tttgtcattt gaaaggaaaa attaacagtt cctataagat acttttgagg tggaatctga 7680

catcctaatt ttttttcttt tcagtgggtt tgcagcgagg gtcttgtatg cactaggcaa 7740

gggttctacc actaagccac atttcccagg aaataaaatg ttaacagtta aaacatacac 7800

acaaatacac aaacacctta ttaccacttt agtaaagtga gagatgtgcg tcctttgtct 7860

cagtctccac gatttcagct gccccttgta tgaataactc agtctcgcta aactgtttac 7920

ttttatttac ctggtttgac tagttgcagc tatataacca gttgtgcatg aggacaacag 7980

ccagtgtgtt tgttttgttt ttggtttttt gtggtacatt ttttgtaaag aattctgtag 8040

attgaagtgc tctttgaaaa cagaactgag atatatttat tcttgttagc atcaaaaaac 8100

attttgtgca aatgatttgc ttttcctggc aggctgagta ccatatccag cgcccacaat 8160

tgcgggttcc catctaccat gtccacaggg gagacagacg ggaagcacat gaggggtgtg 8220

tttacagagt tgtaggagtt atgtagttct cttgttgcct tggaaatcac tgttgtttta 8280

agactgttga acccgtgtgt ttggctgggc tgtgagttac atgaagaaac tgcaaactag 8340

catatgcaga caaagctcac agactaggcg taaatggagg aaaatggacc aaaataaggc 8400

agggtgacac ataaaccttg ggcttcggag aaaactaagg gtggagatga actataatca 8460

cctgaataca atgtaagagt gcaataagtg tgcttattct aagctgtgaa cttcttttaa 8520

atcattcctt tctaatacat ttatgtatgt tccattgctg actaaaacca gctatgagaa 8580

catatgcctt tttattcatg ttaactacca gtttaagtgg ctaaccttaa tgtcttattt 8640

atcttcattt tgtattagtt tacataccag gtatgtgtgt gtgctgtact cttcttccct 8700

ttatttgaaa acacttttca ctgggtcatc tccttggcca ttccacaaca caactttggt 8760

ttggctttca atgtcacctt atttgatggc ctgtgtccca gtagcagaat ttatggtatt 8820

cccattgctg gctgctcttc cgaccctttg cttctacagc acttgtctct cctaagatag 8880

tcagaaacta actgatcagg ggatggactt caccattcat cgtgtctctt caattctatt 8940

aaatagacca ctcttgggct ttagaccagg aaaaaggaga cagctctagc catctaccaa 9000

gcctcaccct aaaaggtcac ccgtacttct tggtctgagg acaagtctcc actccagtaa 9060

gggagagggg aggaaatgct tcctgtttga aatgcagtga attcctatgg ctcctgtttc 9120

accacccgca cctatggcaa cccatataca ttcctcttgt ctgtaactgc caaaggttgg 9180

gtttatgtca cttcagttcc actcaagcat tgaaaaggtt ctcatggagt ctggggtgtg 9240

cccagtgaaa agatggggac tttttcatta tccacagacc tctctatacc tgctttgcaa 9300

aaattataat ggagtaacta tttttaaagc ttatttttca attcataaga aaaagacatt 9360

tattttcaat caaatggatg atgtctctta tcccttatcc ctcaatgttt gcttgaattt 9420

tgtttgttcc ctatacctac tccctaattc tttagttcct tcctgctcag gtcccttcat 9480

ttgtactttg gagtttttct catgtaaatt tgtataatgg aaaatattgt tcagtttgga 9540

tagaaagcat ggagaaataa ataaaaaaag atagctgaaa atcaaattga agaaatttat 9600

ttctgtgtaa agttatttaa aaactctgta ttatatttaa agaaaaaagc ccaacccccc 9660

aaaaagtgct atgtaattga tgtgaatatg cgaatactgc tataataaag attgactgca 9720

tggagaaa 9728

<210>6

<211>804

<212>PRT

<213> mice

<400>6

Met Ala Ala Leu Ser Gly Gly Gly Gly Ser Ser Ser Gly Gly Gly Gly

1 5 10 15

Gly Gly Gly Gly Gly Gly Gly Gly Gly Asp Gly Gly Gly Gly Ala Glu

20 25 30

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

35 40 45

Gly Ala Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val

50 55 60

Trp Asn Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala

65 70 75 80

Leu Leu Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu

85 90 95

Glu Ala Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg

100 105 110

Glu Gln Gln Leu Leu Glu Ser Leu Val Phe Gln Thr Pro Thr Asp Ala

115 120 125

Ser Arg Asn Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe

130 135 140

Leu Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr

145 150 155 160

Val Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro

165 170 175

Glu Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile

180 185 190

Gly Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val

195 200 205

Glu Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys

210 215 220

Thr Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe

225 230 235 240

Gln Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys

245 250 255

Ser Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu

260 265 270

Leu Phe Val Ser Lys Phe Phe Glu His His Pro Val Pro Gln Glu Glu

275 280 285

Ala Ser Phe Pro Glu Thr Ala Leu Pro Ser Gly Ser Ser Ser Ala Pro

290 295 300

Pro Ser Asp Ser Thr Gly Pro Gln Ile Leu Thr Ser Pro Ser Pro Ser

305 310 315 320

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

325 330 335

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

340 345 350

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Glu Lys Phe Pro Glu

355 360 365

Val Glu Leu Gln Asp Gln Arg Asp Leu Ile Arg Asp Gln Gly Phe Arg

370 375 380

Gly Asp Gly Ala Pro Leu Asn Gln Leu Met Arg Cys Leu Arg Lys Tyr

385 390 395 400

Gln Ser Arg Thr Pro Ser Pro Leu Leu His Ser Val Pro Ser Glu Ile

405 410 415

Val Phe Asp Phe Glu Pro Gly Pro Val Phe Arg Gly Ser Thr Thr Gly

420 425 430

Leu Ser Ala Thr Pro Pro Ala Ser Leu Pro Gly Ser Leu Thr Asn Val

435 440 445

Lys Ala Leu Gln Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser

450 455 460

Ser Ser Ser Ser Ser Glu Asp Arg Ser Arg Met Lys Thr Leu Gly Arg

465 470 475 480

Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly Gln Ile Thr Val

485 490 495

Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys

500505 510

Trp His Gly Asp Val Ala Val Lys Met Leu Asn Val Thr Ala Pro Thr

515 520 525

Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val Gly Val Leu Arg Lys

530 535 540

Thr Arg His Val Asn Ile Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro

545 550 555 560

Gln Leu Ala Ile Val Thr Gln Trp Cys Glu Gly Ser Ser Leu Tyr His

565 570 575

His Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys Leu Ile Asp

580 585 590

Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His Ala Lys Ser

595 600 605

Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe Leu His Glu Asp

610 615 620

Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr Val Lys Ser Arg

625 630 635 640

Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp

645 650 655

Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe

660 665 670

Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu Tyr Glu Leu Met Thr

675 680 685

Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe

690 695 700

Met Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys Val Arg Ser

705 710 715 720

Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys Leu Lys Lys

725 730 735

Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Ile Leu Ala Ser Ile Glu

740 745 750

Leu Leu Ala Arg Ser Leu Pro Lys Ile His Arg Ser Ala Ser Glu Pro

755 760 765

Ser Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala

770 775 780

Cys Ala Ser Pro Lys Thr Pro Ile Gln Ala Gly Gly Tyr Gly Glu Phe

785 790 795 800

Ala Ala Phe Lys

<210>7

<211>2232

<212>DNA

<213> Rabbit (Oryctolagus cuniculus)

<400>7

atggggaatg tgtggaatat caaacaaatg attaagttga cacaggagca tatagaggcc 60

ctattggaca aatttggtgg ggagcataat ccaccatcaa tatatctgga ggcctacgaa 120

gaatacacca gcaagctaga tgccctccaa caaagagaac agcagttatt ggaatcccta 180

gtttttcaaa atcccacaga tgtgtcacgg agcaacccca agtcaccaca aaaacctatt 240

gttagagtct tcctgcccaa caaacagagg acagtggtac ctgcaagatg tggagttacg 300

gttcgagaca gtctaaagaa agcgctgatg atgagaggtc tgatcccaga atgctgtgct 360

gtttacagaa ttcaggatgg agagaagaag ccaattggct gggacactga tatttcctgg 420

ctcactggag aagagctgca tgtggaagtg ttagagaatg tcccactcac cacacataac 480

tttgtacgga aaactttttt caccttagca ttttgtgact tctgtagaaa gctgcttttc 540

cagggtttcc gctgtcaaac atgtggctac aaatttcacc agcgttgtag tacggaagtt 600

ccactgatgt gtgttaatta tgaccaactt gatttgctgt ttgtctccaa gttctttgaa 660

caccacccag taccacagga ggaggcctcc ttagcagaga ctgccctcac atctgggtca 720

tcgccttccg cacctccctc agactctatt gggcaccaaa ttctcaccag tccgtcccct 780

tcaaaatcca ttccgattcc acagtccttc cgaccagcag atgaagatca tcgaaatcag 840

tttgggcaac gagaccggtc ttcatcagcg cctaatgttc acattaacac aatagaacct 900

gtcaatattg atgaaaaatt cccagaagtg gaattacagg atcaaaggga cttgattaga 960

gaccaagggt ttcgtggtga tggagcccct ttgaaccagc tgatgcgctg tcttcggaaa 1020

taccaatccc ggactcccag tcccctccta ccttctgtcc ccagtgacat agtgtttgat 1080

tttgagcctg gcccagtgtt cagaggatcg accacgggtt tgtctgccac tccccctgcc 1140

tcattacctg gctcactcac tagtgtgaaa gctgtacaga gatccccagg acctcagcga 1200

gagaggaagt cgtcttcctc ctcagaagac aggaatcgaa tgaaaactct tggtagacgg 1260

gattcaagtg atgattggga gattcctgat gggcagatca ccgtgggaca gagaattgga 1320

tctggatcat ttggaaccgt ctacaaggga aaatggcacg gtgatgtggc agtaaaaatg 1380

ttgaatgtga cagcacctac acctcagcag ttacaggcct tcaaaaatga agtaggagta 1440

ctcaggaaaa cacgacatgt gaatatccta cttttcatgg gctattccac aaagccacag 1500

ctggctattg ttacccagtg gtgtgagggc tccagtttat atcaccatct ccacatcatt 1560

gagaccaaat tcgagatgat caaacttata gatattgcac ggcagactgc acagggcatg 1620

gattacttac acgccaagtc aatcatccac agagacctca agagtaataa tatatttctt 1680

catgaagacc tcacagtaaa aataggtgat tttggtctag ccacagtgaa atctcgatgg 1740

agtgggtccc atcagtttga acaattgtct ggatccattt tgtggatggc accagaagta 1800

atcagaatgc aagacaaaaa cccatatagc tttcagtcag atgtatatgc atttgggatt 1860

gttctgtatg aattgatgac tgggcagtta ccttactcaa acatcaacaa cagggaccag 1920

atcattttta tggtgggacg tggctacctg tctccagacc tcagtaaggt acggagtaac 1980

tgtccgaaag ccatgaagag attaatggca gagtgcctca aaaagaaaag agatgagaga 2040

ccactctttc cccaaattct cgcctccatt gagctgctgg cccgctcatt gccaaaaatc 2100

caccgcagtg catcagaacc ctccttgaat cgggctggtt tccagacaga ggattttagt 2160

ctatatgctt gtgcttctcc aaaaacaccc atccaggcag ggggatatgg agaatttgca 2220

gccttcaagt ag 2232

<210>8

<211>743

<212>PRT

<213> Rabbit

<400>8

Met Gly Asn Val Trp Asn Ile Lys Gln Met Ile Lys Leu Thr Gln Glu

1 5 10 15

His Ile Glu Ala Leu Leu Asp Lys Phe Gly Gly Glu His Asn Pro Pro

20 25 30

Ser Ile Tyr Leu Glu Ala Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala

35 40 45

Leu Gln Gln Arg Glu Gln Gln Leu Leu Glu Ser Leu Val Phe Gln Asn

50 55 60

Pro Thr Asp Val Ser Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile

65 70 75 80

Val Arg Val Phe Leu Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg

85 90 95

Cys Gly Val Thr Val Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg

100 105 110

Gly Leu Ile Pro Glu Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu

115 120 125

Lys Lys Pro Ile Gly Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu

130 135 140

Glu Leu His Val Glu Val Leu Glu Asn Val Pro Leu Thr Thr His Asn

145 150 155 160

Phe Val Arg Lys Thr Phe Phe ThrLeu Ala Phe Cys Asp Phe Cys Arg

165 170 175

Lys Leu Leu Phe Gln Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe

180 185 190

His Gln Arg Cys Ser Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp

195 200 205

Gln Leu Asp Leu Leu Phe Val Ser Lys Phe Phe Glu His His Pro Val

210 215 220

Pro Gln Glu Glu Ala Ser Leu Ala Glu Thr Ala Leu Thr Ser Gly Ser

225 230 235 240

Ser Pro Ser Ala Pro Pro Ser Asp Ser Ile Gly His Gln Ile Leu Thr

245 250 255

Ser Pro Ser Pro Ser Lys Ser Ile Pro Ile Pro Gln Ser Phe Arg Pro

260 265 270

Ala Asp Glu Asp His Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser

275 280 285

Ser Ala Pro Asn Val His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp

290 295 300

Glu Lys Phe Pro Glu Val Glu Leu Gln Asp Gln Arg Asp Leu Ile Arg

305 310 315 320

Asp Gln Gly Phe Arg Gly Asp Gly Ala Pro Leu Asn Gln Leu Met Arg

325 330 335

Cys Leu Arg Lys Tyr Gln Ser Arg Thr Pro Ser Pro Leu Leu Pro Ser

340 345 350

Val Pro Ser Asp Ile Val Phe Asp Phe Glu Pro Gly Pro Val Phe Arg

355 360 365

Gly Ser Thr Thr Gly Leu Ser Ala Thr Pro Pro Ala Ser Leu Pro Gly

370 375 380

Ser Leu Thr Ser Val Lys Ala Val Gln Arg Ser Pro Gly Pro Gln Arg

385 390 395 400

Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn Arg Met Lys Thr

405 410 415

Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly Gln

420 425 430

Ile Thr Val Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr Val Tyr

435440 445

Lys Gly Lys Trp His Gly Asp Val Ala Val Lys Met Leu Asn Val Thr

450 455 460

Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val Gly Val

465 470 475 480

Leu Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met Gly Tyr Ser

485 490 495

Thr Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu Gly Ser Ser

500 505 510

Leu Tyr His His Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys

515 520 525

Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His

530 535 540

Ala Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe Leu

545 550 555 560

His Glu Asp Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr Val

565 570 575

Lys Ser Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser Gly Ser

580 585 590

Ile Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys Asn Pro

595 600 605

Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu Tyr Glu

610 615 620

Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln

625 630 635 640

Ile Ile Phe Met Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys

645 650 655

Val Arg Ser Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys

660 665 670

Leu Lys Lys Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Ile Leu Ala

675 680 685

Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro Lys Ile His Arg Ser Ala

690 695 700

Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp Phe Ser

705 710 715 720

Leu Tyr Ala Cys Ala Ser Pro LysThr Pro Ile Gln Ala Gly Gly Tyr

725 730 735

Gly Glu Phe Ala Ala Phe Lys

740

<210>9

<211>2186

<212>DNA

<213> Guinea pig (Cavia porcellus)

<400>9

atggcggcgc tcagcggcgg cggtggcgcg gagcagggcc aggctctgtt caacggggac 60

atggagctcg aggccggcgc cggcgccgca gcctcttcgg ctgcagaccc tgccattccc 120

gaggaggtat ggaatatcaa acaaatgatt aagttgacgc aggaacacat agaggcccta 180

ttggacaaat ttggtggaga gcataatcca ccatcaatat acctggaggc ctatgaagaa 240

tacaccagca aactagatgc cctccaacaa agagaacagc agttactgga atccctcggg 300

aatggaactg atttttctgt ttctagctct gcatcactgg acaccgttac atcttcttct 360

tcttctagcc tttcagtact accttcatct ctttcagttt ttcaaaatcc tacagatgtg 420

tcacggagca accccaaatc accacaaaaa cctattgtta gagtcttcct gcccaacaaa 480

cagaggacag tggtacctgc aaggtgtgga gttacagtcc gagacagtct gaagaaagca 540

ctcatgatga gaggtcttat cccagagtgc tgtgctgtgt acagaattca ggatggagaa 600

aagaaaccaa ttggctggga cactgacatttcctggctta ctggggaaga attacatgta 660

gaagtattgg agaatgttcc acttacaaca cacaattttg tatgtatctt tatatttttt 720

ttgctgtttg tctccaagtt ctttgaacac cacccaatac cacaggagga ggcttcctta 780

gcagagacca cccttacatc tggatcatcc ccttctgcac ccccctcaga gtccattggg 840

cccccaattc tcaccagccc atctccttca aaatccattc caattccaca gcctttccgg 900

ccaggagagg aagatcatcg aaatcaattt gggcagcgag accggtcctc atctgctccc 960

aatgtgcata taaacacaat agaacctgtc aatattgatg atttgattag agaccaaggg 1020

tttcgtagtg atggaggatc aactacaggt ttgtctgcca ccccacctgc ctcattacct 1080

ggctcactca ctaatgtgaa agccttacag aaatctccag gacctcagcg agaaaggaag 1140

tcatcttcat cctcagaaga cagaaatcga atgaaaacgc ttggtagacg ggactcaagt 1200

gatgattggg agattcctga tgggcagatt acagtgggac aaagaattgg atctgggtca 1260

tttggaacag tctacaaggg gaagtggcat ggtgacgtgg cagtgaaaat gttgaatgtg 1320

acagcaccca cacctcaaca gttacaggcc ttcaaaaatg aagtaggagt actcaggaaa 1380

acacgacatg tgaatatcct actcttcatg ggctattcca caaagccaca gctagctatt 1440

gttacccagt ggtgtgaggg ctccagctta taccaccatc tccacatcat cgagaccaaa 1500

tttgagatga tcaaacttat agatattgca cgacagactg cccagggcat ggattactta 1560

cacgccaagt caatcatcca cagagacctc aagagtaata atatatttct tcacgaagac 1620

ctcacggtta aaataggtga ttttggtcta gccacagtga aatctcgatg gagtgggtcc 1680

catcagtttg aacagttgtc tggatccatt ttgtggatgg caccagaagt aatcagaatg 1740

cgagataaaa acccatacag ttttcagtcc gatgtatatg catttgggat tgttctatat 1800

gaattgatga ctgggcagtt accctattca aatatcaaca acagggacca gataattttt 1860

atggtgggac gaggatatct atctccagat ctcagcaagg tacggagtaa ctgtccaaaa 1920

gccatgaaga ggttaatggc ggagtgcctc aaaaagaaaa gagatgagag accactcttt 1980

ccccaaattc tcgcctctat tgagctgctg gcccgctcat tgccaaaaat tcaccgcagt 2040

gcatcagaac cctccttgaa tcgggctggt ttccaaacag aggattttag tctctatgct 2100

tgtgcttctc caaaaacacc catccaggca gggggatatg gtgcgtttcc tgtccactga 2160

tgcaaattaa atgagtgaga aataaa 2186

<210>10

<211>719

<212>PRT

<213> Guinea pig

<400>10

Met Ala Ala Leu Ser Gly Gly Gly Gly Ala Glu Gln Gly Gln Ala Leu

1 5 10 15

Phe Asn Gly Asp Met Glu Leu Glu Ala Gly Ala Gly Ala Ala Ala Ser

20 25 30

Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn Ile Lys Gln

35 40 45

Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu Asp Lys Phe

50 55 60

Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala Tyr Glu Glu

65 70 75 80

Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln Gln Leu Leu

85 90 95

Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser Ser Ala Ser

100 105 110

Leu Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser Val Leu Pro

115 120 125

Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser Arg Ser Asn

130 135 140

Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu Pro Asn Lys

145 150 155 160

Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val Arg Asp Ser

165 170 175

Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys Cys Ala

180 185 190

Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly Trp Asp Thr

195 200 205

Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu Val Leu Glu

210 215 220

Asn Val Pro Leu Thr Thr His Asn Phe Val Cys Ile Phe Ile Phe Phe

225 230 235 240

Leu Leu Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu

245 250 255

Glu Ala Ser Leu Ala Glu Thr Thr Leu Thr Ser Gly Ser Ser Pro Ser

260 265 270

Ala Pro Pro Ser Glu Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser

275 280 285

Pro Ser Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Gly Glu Glu

290 295 300

Asp HisArg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro

305 310 315 320

Asn Val His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile

325 330 335

Arg Asp Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser

340 345 350

Ala Thr Pro Pro Ala Ser Leu Pro Gly Ser Leu Thr Asn Val Lys Ala

355 360 365

Leu Gln Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser

370 375 380

Ser Glu Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser

385 390 395 400

Asp Asp Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile

405 410 415

Gly Ser Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp

420 425 430

Val Ala Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu

435 440 445

Gln Ala Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val

450 455 460

Asn Ile Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile

465 470 475 480

Val Thr Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile

485 490 495

Ile Glu Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln

500 505 510

Thr Ala Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg

515 520 525

Asp Leu Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys

530 535 540

Ile Gly Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser

545 550 555 560

His Gln Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu

565 570 575

Val Ile Arg Met Arg Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val

580 585 590

Tyr Ala Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro

595 600 605

Tyr Ser Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg

610 615 620

Gly Tyr Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys

625 630 635 640

Ala Met Lys Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu

645 650 655

Arg Pro Leu Phe Pro Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg

660 665 670

Ser Leu Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg

675 680 685

Ala Gly Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro

690 695 700

Lys Thr Pro Ile Gln Ala Gly Gly Tyr Gly Ala Phe Pro Val His

705 710 715

<210>11

<211>3229

<212>DNA

<213> dog (Canis familiaris)

<400>11

gtaatgctgg attttcatgg aataagtttg acctgtgctg cagtggcctc cagcaaggta 60

cccgcaagat gtggagttac agtccgggac agtctaaaga aagctctgat gatgagaggt 120

ctaatcccag agtgctgtgc tgtttacaga attcaggatg gagagaagaa accgattggc 180

tgggacactg atatttcctg gctcactgga gaggaattgc atgtagaagt gttggaaaat 240

gttccgctta ccacacacaa ctttgtacgg aaaacttttt tcaccttagc attttgtgac 300

ttttgtcgaa agctgctttt ccagggtttt cgctgtcaaa catgtggtta taaatttcac 360

cagcgttgta gtacagaggt tccactgatg tgtgttaatt atgaccaact tgatttgctg 420

tttgtctcca agttctttga acaccaccca ataccacagg aggaggcctc catagcagag 480

actgccctta cgtctggatc atccccttct gctcccccct ccgattctcc tgggccccca 540

attctgacca gtccgtctcc ttcaaaatcc attccaattc cacagccttt ccgaccagca 600

gatgaagatc atcgaaatca gtttggacaa cgagaccggt cctcatcagc tccaaatgtg 660

catataaaca caatagaacc cgtcaacatt gatgacttga ttagagacca agggtttcgt 720

agtgatggag gatcaaccac aggtttgtct gccacccccc ctgcctcatt gcctggctca 780

ctcactaatg taaaagcatt acagaaatct ccaggacctc agcgggaaag aaaatcatct 840

tcatcctcag aagataggaa tcgaatgaaa acacttggta gacgggattc aagtgatgat 900

tgggagatac ctgatgggca gatcacagtg ggacagagaa ttggatccgg gtcatttggg 960

acagtctaca agggaaagtg gcatggtgac gtggcagtga aaatgttgaa tgtgacagca 1020

cccacacctc agcagttaca ggccttcaaa aatgaagtag gagtactcag gaaaactcga 1080

catgtgaata tcctactctt tatgggctat tcaacaaagc cccaactggc tattgttacc 1140

cagtggtgtg agggctccag cttatatcac catctccaca tcattgagac caaatttgag 1200

atgataaagc ttatagatat tgcacggcag actgcacagg gcatggatta cttacacgcc 1260

aagtcaatca tccacagaga cctcaagagt aataatattt ttcttcatga agacctcaca 1320

gtaaaaatag gtgattttgg tctagccaca gtgaaatctc gatggagtgg gtcccatcag 1380

tttgaacagt tgtctggatc cattttgtgg atggcaccag aagtgatccg aatgcaagac 1440

aaaaacccat atagcttcca gtcagatgta tacgcatttg ggattgttct atatgaattg 1500

atgacagggc agttacctta ttcaaacatc aacaacaggg accagataat ttttatggtg 1560

ggacgaggat atctttctcc agatctcagt aaggtacgga gtaactgtcc aaaagccatg 1620

aagagattga tggcagagtg cctaaaaaag aaaagagatg agaggccact ctttccccaa 1680

attctcgcct ctattgagct gctggcccgc tcattgccaa aaattcaccg cagtgcatca 1740

gaaccctcct tgaatcgggc tggcttccaa acagaggatt ttagtctcta tgcttgcgct 1800

tctccaaaaa cacccatcca ggcaggggga tacggagaat ttgcagcctt caagtagcca 1860

caccatcatg gcaacaacta ctcttatttc ttaagtcttg tgttcgtaca atttgttaac 1920

atcaaaacac agttctgttc ctcaaatctt tttttaaaga tacagaattt tcaatgcata 1980

agctggtgtg gaacagaatg gaatttccca tccaacaaaa gagggaagaa tgttttagga 2040

accagaattc tctgctgcca gtgtttcttc ttcaacacaa ataccacgtg catacaagtc 2100

tgcccactcc caggaaggaa gaggagagcc tgagttctga ccttttgatg gtcaggcatg 2160

atggaaagaa actgctgcta cagcttggga gattggctgt ggagagcctg cccgtcagct 2220

ctgcccttct aaccgccaga tgagtgtgtg gctggtcacc tgacagggca gctgcaatcg 2280

ccaagcatcg ttctctttcc tgtcctggga ttttgtcgtg gagctctttc cccctagtca 2340

ccaccggttc atttctgagg gatggaacaa aaatgcagca tggcctttct gtgtggtgca 2400

tgtccggtct ttgacaaatt tttatcaagt gaagctcttg tatttaaatg gagaatgaga 2460

ggcgaggggg ggggatcacg ttttggtgta ggggcaaagg gaatgctgca tctttttcct 2520

gacccactgg gtttctggcc tttgtttcct tgctcactga gggtgtctgc ctataaccac 2580

gcaggctgga aagtgctggc acacattgcc ttctcttctc actgggtcca gcaatgaaga 2640

caagtgttgg ggattttttt ttttgccctc cacaatgtag caagttctca ggaaaataca 2700

gttaatatct tcctcctaag ctcttccagt catcaagtac ttatgtggct actttgtcca 2760

gggcacaaaa tgccatggcg gtatccaatt aaaagcctac aaaactgctt gataacagtt 2820

ttgaatgtgt gagacattta tgtaatttaa atgtaaggta caagttttaa tttctgagtt 2880

tctctattat atttttatta aaaagaaaat aattttcaga tttaattgaa ttggaataaa 2940

ataatacttc ccaccagaat tatatatcct ggaaaattgt atttttgtta tataaacaac 3000

ttttaaagaa agatcattat ccttttctct acctaaatat ggggagtctt agcataatga 3060

cagatattta taatttttaa attaatggta cttgctggat ccacactaac atctttgcta 3120

atatctcatg ttttcctcca acttactcct acactacatc ctccatcctc tttccagtct 3180

tttatctaga atatgcaacc taaaataaaa atggtggtgt ctccattca 3229

<210>12

<211>617

<212>PRT

<213> dog

<400>12

Met Leu Asp Phe His Gly Ile Ser Leu Thr Cys Ala Ala Val Ala Ser

1 5 10 15

Ser Lys Val Pro Ala Arg Cys Gly Val Thr Val Arg Asp Ser Leu Lys

20 25 30

Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys Cys Ala Val Tyr

35 4045

Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly Trp Asp Thr Asp Ile

50 55 60

Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu Val Leu Glu Asn Val

65 70 75 80

Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr Phe Phe Thr Leu Ala

85 90 95

Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln Gly Phe Arg Cys Gln

100 105 110

Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser Thr Glu Val Pro Leu

115 120 125

Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu Phe Val Ser Lys Phe

130 135 140

Phe Glu His His Pro Ile Pro Gln Glu Glu Ala Ser Ile Ala Glu Thr

145 150 155 160

Ala Leu Thr Ser Gly Ser Ser Pro Ser Ala Pro Pro Ser Asp Ser Pro

165 170 175

Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser Lys Ser Ile Pro Ile

180 185 190

Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His Arg Asn Gln Phe Gly

195 200 205

Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val His Ile Asn Thr Ile

210 215 220

Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp Gln Gly Phe Arg Ser

225 230 235 240

Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr Pro Pro Ala Ser Leu

245 250 255

Pro Gly Ser Leu Thr Asn Val Lys Ala Leu Gln Lys Ser Pro Gly Pro

260 265 270

Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn Arg Met

275 280 285

Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp

290 295 300

Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr

305 310 315 320

Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala Val Lys Met LeuAsn

325 330 335

Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val

340 345 350

Gly Val Leu Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met Gly

355 360 365

Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu Gly

370 375 380

Ser Ser Leu Tyr His His Leu His Ile Ile Glu Thr Lys Phe Glu Met

385 390 395 400

Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr

405 410 415

Leu His Ala Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile

420 425 430

Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala

435 440 445

Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser

450 455 460

Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys

465 470 475 480

Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu

485 490 495

Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn Arg

500 505 510

Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu

515 520 525

Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala

530 535 540

Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Ile

545 550 555 560

Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro Lys Ile His Arg

565 570 575

Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp

580 585 590

Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys Thr Pro Ile Gln Ala Gly

595 600 605

Gly Tyr Gly Glu Phe Ala Ala Phe Lys

610 615

<210>13

<211>1889

<212>DNA

<213> dog (Canis familiaris)

<400>13

ggaatatcaa acaaatgatt aagttgacac aggaacatat agaagcccta ttggacaagt 60

ttggtgggga gcataatcca ccatcaatat atctggaggc ctatgaagaa tacaccagca 120

aactagatgc cctccaacag cgagaacaac agttattgga atccctgggg aatggaactg 180

atttttctgt ttctagctct gcatcaacgg acaccgttac atcttcttcc tcttctagcc 240

tttcagtgct accttcatct ctttcagttt ttcaaaatcc cacagatata tcacggagca 300

atcccaagtc accacaaaaa cctatcgtta gagtcttcct gcccaataaa cagaggacgg 360

tggtacccgc aagatgtgga gttacagtcc gggacagtct aaagaaagct ctgatgatga 420

gaggtctaat cccagagtgc tgtgctgttt acagaattca ggatggagag aagaaaccga 480

ttggctggga cactgatatt tcctggctca ctggagagga attgcatgta gaagtgttgg 540

aaaatgttcc gcttaccaca cacaactttg tacggaaaac ttttttcacc ttagcatttt 600

gtgacttttg tcgaaagctg cttttccagg gttttcgctg tcaaacatgt ggttataaat 660

ttcaccagcg ttgtagtaca gaggttccac tgatgtgtgt taattatgac caacttgatt 720

tgctgtttgt ctccaagttc tttgaacacc acccaatacc acaggaggag gcctccatag 780

cagagactgc ccttacgtct ggatcatccc cttctgctcc cccctccgat tctcctgggc 840

ccccaattct gaccagtccg tctccttcaa aatccattcc aattccacag cctttccgac 900

cagcagatga agatcatcga aatcagtttg gacaacgaga ccggtcctca tcagctccaa 960

atgtgcatat aaacacaata gaacccgtca acattgatga cttgattaga gaccaagggt 1020

ttcgtagtga tggaggatca accacaggtt tgtctgccac cccccctgcc tcattgcctg 1080

gctcactcac taatgtaaaa gcattacaga aatctccagg acctcagcgg gaaagaaaat 1140

catcttcatc ctcagaagat aggaatcgaa tgaaaacact tggtagacgg gattcaagtg 1200

atgattggga gatacctgat gggcagatca cagtgggaca gagaattgga tccgggtcat 1260

ttgggacagt ctacaaggga aagtggcatg gtgacgtggc agtgaaaatg ttgaatgtga 1320

cagcacccac acctcagcag ttacaggcct tcaaaaatga agtaggagta ctcaggaaaa 1380

ctcgacatgt gaatatccta ctctttatgg gctattcaac aaagccccaa ctggctattg 1440

ttacccagtg gtgtgagggc tccagcttat atcaccatct ccacatcatt gagaccaaat 1500

ttgagatgat aaagcttata gatattgcac ggcagactgc acagggcatg gattacttac 1560

acgccaagtc aatcatccac agagacctca agagtaataa tatttttctt catgaagacc 1620

tcacagtaaa aataggtgat tttggtctag ccacagtgaa atctcgatgg agtgggtccc 1680

atcagtttga acagttgtct ggatccattt tgtggatggc accagaagtg atccgaatgc 1740

aagacaaaaa cccatatagc ttccagtcag atgtatacgc atttgggatt gttctatatg 1800

aattgatgac agggcagtta ccttattcaa acatcaacaa cagggaccag ctcagatcat 1860

gatcacggtg tcatgagatc aagccccac 1889

<210>14

<211>615

<212>PRT

<213> dog

<400>14

Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu Asp Lys Phe

1 5 10 15

Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala Tyr Glu Glu

20 25 30

Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln Gln Leu Leu

35 40 45

Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser Ser Ala Ser

50 55 60

Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser Val Leu Pro

65 70 75 80

Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Ile Ser Arg Ser Asn

85 90 95

Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu Pro Asn Lys

100 105 110

Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val Arg Asp Ser

115 120 125

Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys Cys Ala

130 135 140

Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly Trp Asp Thr

145 150 155 160

Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu Val Leu Glu

165 170 175

Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr Phe Phe Thr

180 185 190

Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln Gly Phe Arg

195 200 205

Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser Thr Glu Val

210215 220

Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu Phe Val Ser

225 230 235 240

Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala Ser Ile Ala

245 250 255

Glu Thr Ala Leu Thr Ser Gly Ser Ser Pro Ser Ala Pro Pro Ser Asp

260 265 270

Ser Pro Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser Lys Ser Ile

275 280 285

Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His Arg Asn Gln

290 295 300

Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val His Ile Asn

305 310 315 320

Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp Gln Gly Phe

325 330 335

Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr Pro Pro Ala

340 345 350

Ser Leu Pro Gly Ser Leu Thr Asn Val Lys Ala Leu Gln Lys Ser Pro

355 360 365

Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn

370 375 380

Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile

385 390 395 400

Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser Gly Ser Phe

405 410 415

Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala Val Lys Met

420 425 430

Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn

435 440 445

Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe

450 455 460

Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys

465 470 475 480

Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu Thr Lys Phe

485 490 495

Glu MetIle Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met

500 505 510

Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn

515 520 525

Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly Asp Phe Gly

530 535 540

Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln Phe Glu Gln

545 550 555 560

Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln

565 570 575

Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile

580 585 590

Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn

595 600 605

Asn Arg Asp Gln Leu Arg Ser

610 615

<210>15

<211>3521

<212>DNA

<213> Cat (Felis cat)

<220>

<221>misc_feature

<222>(630)..(649)

<223> n is a, c, g, or t

<400>15

atgcctaacc tcagtctctg ccaccacggc caatttgctc atgtgcccac tgtgtcggca 60

ctgggatatt ttgtgatttg ccttggccat tgtccactgt ccttgacatt gcctgaagga 120

gaaccactga tgctaatgtt gaaggtgacc tttgcaggct ctccactact cataccaaag 180

atgcggcccc ctgataatcc cagagccact gtctgcacat gggcaaaaca ggctacattc 240

tgtgcagact ggggagaaag gttccaagaa cacagtgcca tagttttggg cagagagttt 300

caacacagca tagtgtctat ggcagtatct ggatttggcc gggggaagtg cccaagagga 360

gacagtcagg ctgtgtccta cggccaagga cctgcactta tttttgcatg cagtggttta 420

gcacagggaa gagaacgaag taggaaatcg gagccatgga aacggcagag cggaggaaac 480

gtgcacgcgc gagggtgggc acgaaaggaa agaaccctcc ccagaagact gcgcgagggc 540

gctcctagga ttacgtcacg caccccgcga aaactgaaat gtactgtgtg tggtctttta 600

attgaactat cttccttatg tgcacttaan nnnnnnnnnn nnnnnnnnng cggcggcggc 660

ggtggcgcgg agcagggcca ggctctgttc aacggggaca tggagcccga agccggcgcc 720

gcggcctctt cggctgcgga ccctgccatt cccgaggagg tgtggaatat caaacaaatg 780

attaagttga cacaggaaca tatagaggcc ctattggaca aatttggtgg ggagcataat 840

ccaccatcaa tatatctaga ggcctatgaa gaatacacca gcaagctaga tgccctccaa 900

cagagagaac aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc 960

tctgcatcaa cagacaccgt tacatcttcc tcctcttcta gcctttcagt gctaccttca 1020

tctctttcag tttttcaaaa ccccacagat gtgtcacgga gcaatcccaa gtcaccacag 1080

aaacctatcg ttagagtctt cctgcctaat aaacagagga cagtggtacc tgcaagatgt 1140

ggagttacag tccgggacag tctaaagaaa gctctgatga tgagaggtct aatccctgag 1200

tgctgtgctg tttacagaat tcaggatgga gagaagaaac caattggctg ggacactgat 1260

atctcctggc tcaccggaga ggaattgcat gtagaagtgt tggaaaatgt tccacttaca 1320

actcacaact ttgtatgtac ggaaaacgtt ttcaccttag cattttgtga cttttgtcga 1380

aagctgcttt tccaaggttt tcgctgtcaa acgtgtggtt ataaatttca ccagcgttgt 1440

agtacagagg ttccactgat gtgtgttaat tatgaccaac ttgatttgct gtttgtctcc 1500

aagttctttg aacaccaccc aataccacag gaggaggcct ccatagcaga gactgcccta 1560

acgtctggat cgtccccttc tgcccccccc tccgattcta ctgggcccca aattctcacc 1620

agtccgtctc cttcaaaatc cattccaatt ccacagcctt tccgaccagc agatgaagat 1680

catcgaaatc aatttggaca gcgagaccgg tcctcatcag ctccaaatgt gcatataaat 1740

acaatagaac ctgtcaatat tgatgacttg attagagacc aggggtttcg tagtgatgga 1800

ggatcaacca caggcttgtc tgccaccccc cctgcctcat tgccgggctc tctcactaat 1860

gtaaaagcat tacagaaatc tccagggcct cagcgggaaa ggaaatcttc ttcatcctca 1920

gaagatagga atcgaatgaa aacacttggt agaagggatt caagtgatga ttgggagatt 1980

cctgatgggc agatcacagt gggacagaga attggatccg ggtcatttgg gacagtctac 2040

aagggaaagt ggcatggtga tgtggcagtg aaaatgttga atgtgacagc acccacacct 2100

cagcagttac aggccttcaa aaatgaagta ggagtactca ggaaaactcg gcatgtgaac 2160

atcctgctct tcatgggcta ttcaacaaag ccccagctgg ctattgtcac ccagtggtgt 2220

gagggctcca gcttatacca ccatctccac atcatcgaga ccaaattcga gatgatcaag 2280

ctgatagata ttgctcggca gactgcgcag ggcatggatt acttacacgc caagtcaatc 2340

atccacagag acctcaagag taataatatt tttcttcacg aagacctcac agtaaaaata 2400

ggtgattttg gtctagccac agtgaaatct cgatggagtg ggtcccatca gtttgaacag 2460

ttgtctggat ccattttgtg gatggcacca gaagtaattc gaatgcaaga taaaaaccca 2520

tatagctttc agtcagatgt atatgcattt gggattgttc tatatgaatt gatgactgga 2580

cagttacctt attcaaacat caacaacagg gaccagataa tttttatggt gggacgagga 2640

tatctttctc cagatctcag taaggtacga agtaactgtc caaaagccat gaagagattg 2700

atggcagagt gcctaaaaaa gaaaagagat gagaggccac tgtttcccca aattcttgcc 2760

tctattgagc tgctggcccg ctcattgcca aaaattcacc gcagtgcatc agaaccctcc 2820

ttgaatcggg ctggcttcca gacagaggat tttagtctct atgcttgtgc ttctccaaaa 2880

acacccatcc aggcaggggg atatggtgcg tttcccgtcc actgagataa gttagatgag 2940

tgcgcgagtg cagggggccg gggccaagga ggtggaaatg tgcgtgcttc tgtactaagt 3000

tggatagcat cttctttttt aaaaaaagat gaaccaaaga atgtgtatgt ttttaaagac 3060

tagatataat tatttcctga tctaaaatgt atacttagct ttggattttc aatatccaag 3120

ggttttcaaa atgcacagac attgctgaac atttgcagta cctcttctgg aggctttact 3180

tcctgttaca aattggtttt gtttactggc ttatcctaat tattaaactt caattaaact 3240

tttctcctgc accttttgtt atgagctatc acatgtccct tagggactcg caagagcagt 3300

actgcccccg tgtacgggct tgcaggtaga aaggggatga cgggttttaa cacctgtgtg 3360

aggcaaggca gtccgaacag atctcattta ggaagccacg agagttgaat aagttatttt 3420

tattcttagt attttttctg taactacttt ttattataac ttggaaaata tggatgtcct 3480

ttatacacct tagcaataga ctgaatttct ttttataaat t 3521

<210>16

<211>974

<212>PRT

<213> Cat

<220>

<221>misc_feature

<222>(210)..(217)

<223> Xaa can be any naturally occurring amino acid

<400>16

Met Pro Asn Leu Ser Leu Cys His His Gly Gln Phe Ala His Val Pro

1 5 10 15

Thr Val Ser Ala Leu Gly Tyr Phe Val Ile Cys Leu Gly His Cys Pro

20 25 30

Leu Ser Leu Thr Leu Pro Glu Gly Glu Pro Leu Met Leu Met Leu Lys

35 40 45

Val Thr Phe Ala Gly Ser Pro Leu Leu Ile Pro Lys Met Arg Pro Pro

50 55 60

Asp Asn Pro Arg Ala Thr Val Cys Thr Trp Ala Lys Gln Ala Thr Phe

65 70 75 80

Cys Ala Asp Trp Gly Glu Arg Phe Gln Glu His Ser Ala Ile Val Leu

85 90 95

Gly Arg Glu Phe Gln His Ser Ile Val Ser Met Ala Val Ser Gly Phe

100 105110

Gly Arg Gly Lys Cys Pro Arg Gly Asp Ser Gln Ala Val Ser Tyr Gly

115 120 125

Gln Gly Pro Ala Leu Ile Phe Ala Cys Ser Gly Leu Ala Gln Gly Arg

130 135 140

Glu Arg Ser Arg Lys Ser Glu Pro Trp Lys Arg Gln Ser Gly Gly Asn

145 150 155 160

Val His Ala Arg Gly Trp Ala Arg Lys Glu Arg Thr Leu Pro Arg Arg

165 170 175

Leu Arg Glu Gly Ala Pro Arg Ile Thr Ser Arg Thr Pro Arg Lys Leu

180 185 190

Lys Cys Thr Val Cys Gly Leu Leu Ile Glu Leu Ser Ser Leu Cys Ala

195 200 205

Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Gly Gly Gly Gly Ala Glu

210 215 220

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

225 230 235 240

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

245 250 255

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

260 265 270

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

275 280 285

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

290 295 300

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

305 310 315 320

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

325 330 335

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

340 345 350

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

355 360 365

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

370 375 380

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

385 390 395 400

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

405 410 415

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

420 425 430

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Cys Thr Glu

435 440 445

Asn Val Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe

450 455 460

Gln Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys

465 470 475 480

Ser Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu

485 490 495

Leu Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu

500 505 510

Ala Ser Ile Ala Glu Thr Ala Leu Thr Ser Gly Ser Ser Pro Ser Ala

515 520 525

Pro Pro Ser Asp Ser Thr Gly Pro Gln Ile Leu Thr Ser Pro Ser Pro

530 535 540

Ser Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp

545 550 555 560

His Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn

565 570 575

Val His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg

580 585 590

Asp Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala

595 600 605

Thr Pro Pro Ala Ser Leu Pro Gly Ser Leu Thr Asn Val Lys Ala Leu

610 615 620

Gln Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser

625 630 635 640

Glu Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp

645 650 655

Asp Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly

660 665670

Ser Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val

675 680 685

Ala Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln

690 695 700

Ala Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn

705 710 715 720

Ile Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val

725 730 735

Thr Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile

740 745 750

Glu Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr

755 760 765

Ala Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp

770 775 780

Leu Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile

785 790 795 800

Gly Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His

805 810 815

Gln Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val

820 825 830

Ile Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr

835 840 845

Ala Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr

850 855 860

Ser Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly

865 870 875 880

Tyr Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala

885 890 895

Met Lys Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg

900 905 910

Pro Leu Phe Pro Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser

915 920 925

Leu Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala

930 935 940

Gly Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys

945 950 955 960

Thr Pro Ile Gln Ala Gly Gly Tyr Gly Ala Phe Pro Val His

965 970

<210>17

<211>3853

<212>DNA

<213> cattle (Bos taurus)

<400>17

ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60

cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120

ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180

ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240

agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt 300

cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg attaagttga 360

cacaggagca tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa 420

tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac 480

aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540

cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag 600

tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660

ttagagtctt cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720

tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg 780

tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc 840

ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact 900

ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960

agggattccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc 1020

cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080

accacccaat accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat 1140

ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt 1200

caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260

ttggacaacg agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg 1320

tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380

gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440

agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc 1500

gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct gatggacaga 1560

tcacagtgggacaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc 1620

atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag cagttacagg 1680

ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740

tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt 1800

tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt atagatattg 1860

cacggcagac tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc 1920

tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc 1980

tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040

ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt 2100

cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag ttaccttatt 2160

caaatatcaa caacagggac cagataattt ttatggtggg acgaggatat ctgtctccag 2220

atctcagtaa ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc 2280

taaaaaagaa aagagatgaa agaccactct ttccccaaat tctcgcctct attgagctgc 2340

tggcccgctc attgccaaaa attcaccgca gtgcatcaga accctccttg aatcgggctg 2400

gcttccaaac agaggatttt agtctatatg cttgtgcttc tccaaaaaca cccattcagg 2460

cagggggata tggtacgttt cctgttcact gaaacaaacc gagtgagtga cagcatgtag 2520

gagggtagggacaaaagaaa gtgaacaaat gtttgcttat atatttgtta aattgaatag 2580

gattttcttt ttctttaaag gtgaacaaga gaacatgtgt gtttttaaag tttggatata 2640

gttttcttcc cagtctaaaa cccatagtta gcattacatt ttcaacatcg aatttttttt 2700

taattcatag acattgctga aaatttataa taccttttcc agaggcttta cttcccattc 2760

caagtttgtt ttgtttactt ggttagtcta atcattaaac tttaaacttt ccccacctac 2820

cttttgctgt tagctatccc gcatccatta ggggctccaa gaacagcact gtctgcgtgt 2880

gtgtgttggc aggtgggaag ctgatggtaa gttaggctgt gttagtgaag gtaaactgac 2940

caggtctaat taggagtcac tagaattgaa taagcttatt tttattaata ttttttctta 3000

taactatttc tttttgtaat aatttagaaa atataattgt tctttattcc cttacagcag 3060

tataaattat tggtgcaggt aaccaaagat attactgagg agtggcatgt ttgacatgag 3120

tgacatggtt taactttgga tttttagtta atatttcttt atatattaag gatgtcttac 3180

acattataga agtcaaattt actgacaaag gtattgcctc ctcttcctcc ccaaaaacac 3240

agcaaaattc tctgggaact cgtagcattg ttggttttct tttggatgac tatggttgcc 3300

aaacaaccaa gtaattgatt ttttttaaat tattattgct ttagattata ctcacctctc 3360

atgatgcctg ttagcaatca cctttatcca tgtgtcttgt aaaatatctt tcctccttat 3420

attctttgcc caacaagagt ctacttgtta tgaatgagta ctattttctt tttttgattc 3480

cccagtataattagtatgtt tagtgctttc taggacttcc actttcttat gttaaaaaaa 3540

aaaacaaact aatgtggcag tcagtatatt cttactgtga atcagagtct ttactgggaa 3600

tcaaagtgaa agaagcagct gttctgactt cagagtcagc ctagggacca aaaccagcct 3660

cttaaataca ccttcattta ttcagtttgg atttgtgatg attttcatta tagctgacag 3720

ttcaaggtta ttcagtggca cacagatagc atctgcataa atgcctttct tcttgaaaat 3780

aaaggagaaa attgggaaga ctttacacca atagtttagt ctttaagtac cacagataac 3840

acacaccata aat 3853

<210>18

<211>764

<212>PRT

<213> cattle

<400>18

Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu Gln

1 5 10 15

Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala Ala

20 25 30

Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn Ile

35 40 45

Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu Asp

50 5560

Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala Tyr

65 70 75 80

Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln Gln

85 90 95

Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser Ser

100 105 110

Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser Val

115 120 125

Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser Arg

130 135 140

Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu Pro

145 150 155 160

Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val Arg

165 170 175

Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys

180 185 190

Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly Trp

195 200 205

Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu Val

210 215 220

Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr Phe

225 230 235 240

Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln Gly

245 250 255

Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser Thr

260 265 270

Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu Phe

275 280 285

Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala Ser

290 295 300

Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro Pro

305 310 315 320

Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser Lys

325 330 335

Ser Ile Pro Ile Pro Gln ProPhe Arg Pro Ala Asp Glu Asp His Arg

340 345 350

Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val His

355 360 365

Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp Gln

370 375 380

Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr Pro

385 390 395 400

Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln Lys

405 410 415

Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp

420 425 430

Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp

435 440 445

Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser Gly

450 455 460

Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala Val

465 470 475 480

Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe

485 490 495

Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile Leu

500 505 510

Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr Gln

515 520 525

Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu Thr

530 535 540

Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln

545 550 555 560

Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu Lys

565 570 575

Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly Asp

580 585 590

Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln Phe

595 600 605

Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile Arg

610615 620

Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe

625 630 635 640

Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn

645 650 655

Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr Leu

660 665 670

Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met Lys

675 680 685

Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro Leu

690 695 700

Phe Pro Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro

705 710 715 720

Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe

725 730 735

Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys Thr Pro

740 745 750

Ile Gln Ala Gly Gly Tyr Gly Thr Phe Pro Val His

755 760

<210>19

<211>4936

<212>DNA

<213> cattle

<400>19

ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60

cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120

ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180

ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240

agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt 300

cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg attaagttga 360

cacaggagca tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa 420

tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac 480

aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540

cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag 600

tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660

ttagagtctt cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720

tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg 780

tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc 840

ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact 900

ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960

agggattccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc 1020

cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080

accacccaat accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat 1140

ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt 1200

caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260

ttggacaacg agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg 1320

tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380

gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440

agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc 1500

gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct gatggacaga 1560

tcacagtggg acaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc 1620

atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag cagttacagg 1680

ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740

tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt 1800

tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt atagatattg 1860

cacggcagac tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc 1920

tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc 1980

tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040

ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt 2100

cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag ttaccttatt 2160

caaatatcaa caacagggac cagataattt ttatggtggg acgaggatat ctgtctccag 2220

atctcagtaa ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc 2280

taaaaaagaa aagagatgaa agaccactct ttccccaagt aggaaagact ctcctaagca 2340

agagacaaaa ttcagaagtt atcagggaaa aagataagca gattctcgcc tctattgagc 2400

tgctggcccg ctcattgcca aaaattcacc gcagtgcatc agaaccctcc ttgaatcggg 2460

ctggcttcca aacagaggat tttagtctat atgcttgtgc ttctccaaaa acacccattc 2520

aggcaggggg atatgaagca gatttggctc ttacatcaaa taaaaataga gtagaagttg 2580

ggatttagag atttcctgac atgcaagaag gaataagcaa gaaaaaaagg tttgttttcc 2640

ccaaatcata tctattgtct tttacttcta ttttttctta aattttttgt gatttcagag 2700

acatgtagag ttttattgat acctaaacta tgagttcttt tttttttttt tttttcatta 2760

ttttgatttt tttggccaag aggcatatgg gatcttagct tgagaaagca acaattttct 2820

tgatgtcatt ttgggtgagg gcacatattg ctgtgaacag tgtggtgata gccaccaggg 2880

accaaactca cacccgctgc attgaaaggt gaagtcttaa acactggacc agcagagaaa 2940

ttcctactct atgagttctt tttgtcatcc cctccccgca ccctccaccc ccaacctaaa 3000

gtctgatgat gaaatcaaca actattccat tagaagcagt agattctggt agcatgatct 3060

ttagtttgtt agtaagattt tgtgctttgt ggggttgtgt cgttttaagg ctaatattta 3120

agtttgtcaa atagaatgct gttcagattg taaaaatgag taataaacat ctgaagtttt 3180

ttttaagtta tttttaacat ggtatataca gttgagctta gagtttatca ttttctgata 3240

ttctcttact tagtagatga attctagcca ttttttataa agatttctgt taagcaaatc 3300

ctgttttcac atgggcttcc tttaagggat tttagattct gctggatatg gtgactgctc 3360

ataagactgt tgaaaattac ttttaagatg tattagaata cttctgaaaa aaaatagcaa 3420

ccttaaaacc ataagcaaaa gtagtaaggg tgtttataca tttctagagt ccctgtttag 3480

gtaatagcct cctatgattg tactttaaat gttttgctct ccaaggtttt agtaacttgg 3540

ctttttttct aatcagtgcc aaactccccc agttttttta actttaaata tgaggtaata 3600

aatcttttac ccttccttga tcttttgact tataatacct tggtcagttg tttcttaaaa 3660

ggaatcctta aatggaaaga gacaatatca ctgtctgcag ttctgattag tagttttatt 3720

cagaatggaa aaacagatta ttcatttttg aaaattgttc aggggtatgt tcattgttag 3780

gaccttggac tttggagtca gtgcctagct atgcattcca ggtctgccat tttctggctg 3840

tgaaattttg gacaagttac ttaaccactt taaaccccag ctttaagaag taaattaacc 3900

ccagtaaatt aagaagtaat agcagccact tcgtagagtt gttatgaggc tcagatgcag 3960

tgcaaatgtg tataaagtat tcagggagtc acctggtata ctataataga cactagaata 4020

gttgccaata ttatcagcat acaatctgag gattctgtca gccaatcatt agcaatctgt 4080

tgtttgttgg gacatgccag tgttctccag ttgaaatcag tagcaatcta aaaatggata 4140

gattattcct catttaaata gtgtgttcat ataagtgatt gcttggatcc ttatcagaag 4200

ttgctgttac tgaaaaatga taaggctgac taaattgtga tagttgtcag ttactaacca 4260

actcccagaa atgaataaga ggaacctatc tctagttcct agtagaaggt atggacaaaa 4320

tagtaggtga aaaataatgt cttgaacccc caaattaagt aagctttaaa gagtacaata 4380

cctcaaaggg tctttgcggt ttaaaatttg tatgctgaga atgatgttca ttgacatgtg 4440

cctatatgta attttttgat agtttaaaag gtgaaatgaa ctacagatgg gagaggtctg 4500

aattttcttg ccttcagtca aatgtgtaat gtggacatat tatttgacct gtgaatttta 4560

tcttttaaaa aagattaatt cctgcttctt ccttcctaat agttgcatta taataatgaa 4620

aatgagttga taatttgggg ggaaagtatt ctacaaatca accttattat tttaccattg 4680

gtttctgaga aattttgttc atttgaaccg tttatagctt gattagaatc atagcatgta 4740

aaacccaact gagggattat ctgcagactt aatgtagtat tatgtaagtt gtcttctttc 4800

atttcgacct tttttgcttt tgttgttgct agatctgtag tatgtagcta gtcacctttc 4860

agcgaggttt cagcgaggct tttctgtgtc tctaggttat ttgagataac ttttttaaaa 4920

ttagctcttg tcctcc 4936

<210>20

<211>797

<212>PRT

<213> cattle

<400>20

Met Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu

1 5 10 15

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

130 135 140

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

275 280 285

Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala

290 295 300

Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro

305 310 315 320

Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser

325 330 335

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

340 345 350

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

355 360 365

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp

370 375 380

Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr

385 390 395 400

Pro Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln

405 410 415

Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu

420 425 430

Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp

435 440 445

Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser

450 455 460

Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala

465 470 475 480

Val LysMet Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala

485 490 495

Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile

500 505 510

Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr

515 520 525

Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu

530 535 540

Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala

545 550 555 560

Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu

565 570 575

Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly

580 585 590

Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln

595 600 605

Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile

610 615 620

Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala

625 630 635 640

Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser

645 650 655

Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr

660 665 670

Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met

675 680 685

Lys Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro

690 695 700

Leu Phe Pro Gln Val Gly Lys Thr Leu Leu Ser Lys Arg Gln Asn Ser

705 710 715 720

Glu Val Ile Arg Glu Lys Asp Lys Gln Ile Leu Ala Ser Ile Glu Leu

725 730 735

Leu Ala Arg Ser Leu Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser

740 745 750

Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys

755 760 765

Ala Ser Pro Lys Thr Pro Ile Gln Ala Gly Gly Tyr Glu Ala Asp Leu

770 775 780

Ala Leu Thr Ser Asn Lys Asn Arg Val Glu Val Gly Ile

785 790 795

<210>21

<211>4154

<212>DNA

<213> cattle

<400>21

ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60

cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120

ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180

ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240

agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt 300

cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg attaagttga 360

cacaggagca tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa 420

tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac 480

aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540

cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag 600

tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660

ttagagtctt cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720

tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg 780

tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc 840

ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact 900

ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960

agggattccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc 1020

cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080

accacccaat accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat 1140

ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt 1200

caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260

ttggacaacg agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg 1320

tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380

gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440

agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc 1500

gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct gatggacaga 1560

tcacagtggg acaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc 1620

atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag cagttacagg 1680

ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740

tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt 1800

tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt atagatattg 1860

cacggcagac tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc 1920

tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc 1980

tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040

ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt 2100

cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag ttaccttatt 2160

caaatatcaa caacagggac cagataattt ttatggtggg acgaggatat ctgtctccag 2220

atctcagtaa ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc 2280

taaaaaagaa aagagatgaa agaccactct ttccccaagt aggaaagact ctcctaagca 2340

agagacaaaa ttcagaagtt atcagggaaa aagataagca gattctcgcc tctattgagc 2400

tgctggcccg ctcattgcca aaaattcacc gcagtgcatc agaaccctcc ttgaatcggg 2460

ctggcttcca aacagaggat tttagtctat atgcttgtgc ttctccaaaa acacccattc 2520

aggcaggggg atatggctga gcacattgtc catcacccac aagtggctgg ttctcatcgc 2580

agaatctacg tagggaatcg ggcgtgaaat tcacttaaga gatagagcag aggaagtgtt 2640

ctgtttacag gaatggagat gagagttatg agtaagttgc ttagtcagtt ggctttgttt 2700

tgaaaattat tgtgttatat ttgtgttaac ctacttgtgt tttgacagta tatgtcacat 2760

aggaagaaac ctcagactag cataataaca aagctcagac taggcacaga tgtacacaga 2820

atggaccaaa atgggatggg ggaaggtatg ggaataagtc taggggtagg gaaaaattga 2880

tgtgagggtg ggaaataaac tgtaattacc tgaaataaaa tgtaagagtg caataagtgt 2940

gctttttatt ctaagctgtg aatgggtttt ttaaaaaaag cattccttcc caatgcattt 3000

gcctatgttc catagctgat taaaaccagc tatataaaca tatgcctttt tattcatgtt 3060

aattaccaat ataaatggct aacctttacg tcttatttat cttcatgtta tgttagttta 3120

catacaggga tgtgtgtgtg tgtgtatgct ataaattttc cctccttcgt ttaaaaacgc 3180

gtttgttgga tcctctctgt ttccttaggc catgccacag ctcatagtct cagcttggcc 3240

ttcctgtcac ctgatctgaa ggactatcac agtgacgtag ctcgttcatt ggttgtacac 3300

actctaaccc ttttccttgc tcagcaatta ctgtgtcttc taaaacagga gtgtacaacc 3360

atgagattgc aattaattgt ttgacatatg tccctttgaa ttctatttat tagttatgat 3420

tgattgctct ttggtttgga ccaagaaaaa cgaaatccca cctccccacc ttttcactta 3480

tttcttactt tgaggacaat tctgtaagag agaggaaagg gaactccttc atgttttaac 3540

tgcagcaagt taatggccct ggtttacacc aaacattatg gtgattcaca ttcacattcc 3600

tctcctctct tgctgccaga ggtttgggtt ttgttcagtt ctgctcaagc actgaaaaag 3660

ttttcatgga gtctggagag tgcccagtga aaagatggtt tttaattgtc cacagacctt 3720

tctgttcctg ctttgcaaaa attacaaagg agtaactatt tttaaagctt atttttcaat 3780

tcataaaaaa gacatttatt ttcagtcaga tgatgtctcc ttgtccctta atcctcaatg 3840

tttgcttgaa tctttttttt ttttctgatt ttctcccatc cccacttctt gatacttctt 3900

gagttctctt tcctgctcag gtcctttcat ttgtactttg gagttttttc tcatgtaaat 3960

ttgtacaatg gaaaatattg ttcagtttgg atagaacgca tggagaatta aataaaaaag 4020

atagctgaaa ttcagattga aatttatttg tgtaaagtta tttaaaaact ctgtactata 4080

taaaaggcaa aaaaagttct atgtacttga tgtgaatatg cgaatactgc tataataaag 4140

attgactgca tgga 4154

<210>22

<211>781

<212>PRT

<213> cattle

<400>22

Met Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu

1 5 10 15

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

130 135 140

Arg Ser Asn Pro Lys Ser Pro GlnLys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

275 280 285

Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala

290 295 300

Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro

305 310 315 320

Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser

325 330 335

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

340 345 350

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

355 360 365

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp

370 375 380

Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr

385 390 395 400

Pro Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln

405 410 415

Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu

420425 430

Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp

435 440 445

Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser

450 455 460

Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala

465 470 475 480

Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala

485 490 495

Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile

500 505 510

Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr

515 520 525

Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu

530 535 540

Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala

545 550 555 560

Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu

565 570 575

Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly

580 585 590

Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln

595 600 605

Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile

610 615 620

Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala

625 630 635 640

Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser

645 650 655

Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr

660 665 670

Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met

675 680 685

Lys Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro

690 695 700

Leu Phe Pro Gln Val Gly Lys Thr Leu Leu Ser Lys Arg Gln Asn Ser

705 710 715 720

Glu Val Ile Arg Glu Lys Asp Lys Gln Ile Leu Ala Ser Ile Glu Leu

725 730 735

Leu Ala Arg Ser Leu Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser

740 745 750

Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys

755 760 765

Ala Ser Pro Lys Thr Pro Ile Gln Ala Gly Gly Tyr Gly

770 775 780

<210>23

<211>7914

<212>DNA

<213> cattle

<400>23

ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60

cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120

ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180

ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240

agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt 300

cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg attaagttga 360

cacaggagca tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa 420

tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac 480

aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540

cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag 600

tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660

ttagagtctt cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720

tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg 780

tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc 840

ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact 900

ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960

agggattccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc 1020

cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080

accacccaat accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat 1140

ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt 1200

caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260

ttggacaacg agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg 1320

tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380

gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440

agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc 1500

gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct gatggacaga 1560

tcacagtggg acaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc 1620

atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag cagttacagg 1680

ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740

tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt 1800

tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt atagatattg 1860

cacggcagac tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc 1920

tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc 1980

tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040

ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt 2100

cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag ttaccttatt 2160

caaatatcaa caacagggac cagataattt ttatggtggg acgaggatat ctgtctccag 2220

atctcagtaa ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc 2280

taaaaaagaa aagagatgaa agaccactct ttccccaaat tctcgcctct attgagctgc 2340

tggcccgctc attgccaaaa attcaccgca gtgcatcaga accctccttg aatcgggctg 2400

gcttccaaac agaggatttt agtctatatg cttgtgcttc tccaaaaaca cccattcagg 2460

cagggggata tggagaattt gcagccttca agtagccaca ccatcatgac agcatctact 2520

cttatttctt aagtcttgtg ttcgtacaat ttgttaacat caaaacacag ttctgttcct 2580

caactctttt taaagttaaa atttttcagt gcataagctg gtgtggaaca gaaggaaatt 2640

tcccatccaa caaaagaggg aagaatgttt taggaaccag aattctctgc tgccagtgtt 2700

tcttcttcaa cacaaatatc acaagtctgc ccactcccag gaagaaagag gagagaccct 2760

gagttctgac cttttgatgg tcaggcatga tggaaagaaa ctgctgctac agcttgggag 2820

atttgctctg ggaagtctgc cagtcaactt tgcccttcta accaccagat caatatgtgg 2880

ctgatcatct gatggggcag ttgcaatcac caagccttgt tctctttcct gttctgggat 2940

tgtgttgtgg aacccttttc cctagccacc accagttcat ttctgaggga tggaacaaaa 3000

atgcagcatg cccttcctgt gtggtgcatg ttcagtcctt gacaaatttt taccaaaatg 3060

aagctacttt atttaaaagg agggtgagag gtgaggaggt cactttgggt gtggcggaaa 3120

gggaatgctg catctttttc ctgggctgct ggggctctgg ccttggcttg ccagccggaa 3180

gcgctggcac gcatcgcctt cttttcccat tgggtccagc aatgaagacg agtgtttggg 3240

gttttttttt tctccaccat gtagcaagtt ctcaggaaaa tacaattgat atcttcctcc 3300

taagctcttc caatcagtca ccaagtactt atgtggttac tttgtccagg gcacaaaatg 3360

cctgtatcta attaaaagcc tacaaaactg cttgataaca gttttgaatg tgagacattt 3420

atgtaattta aatgtaaggt acaagtttta atttctgagt ttcttctatt atatttttat 3480

taaaaaaaga aaataatttt cagattgaat tggagtaaaa taatattact tcccactaga 3540

attatatatc ctggaaaatt gtatttttgt tacataagca gcttttaaag aaagatcatt 3600

acccttttct ctacataaat atatggggag tcttagccta atgacaaata tttataattt 3660

ttaaattaat ggtacttgct ggatccatac taacatcttt actaatacct cattgtttct 3720

tccaacttac tcctacacta catcctacat cttcttccta gtcttttatc tagaatatgc 3780

aacctcaaat aaaaatggtg gtgtcctcat tcattctcct ccttcctttt ttcccaagcc 3840

tgatcttcaa aaggttggtt aatttggcag ctgagttcct ccccaggcag agaatagacc 3900

aattttaggt gtattgggac tgagggagga tgtgtaaaga ttaacatcag taaagaaccg 3960

ctgtggagta attaagaact ttgttcttta taactggaga atataaccta accctaacat 4020

ccctcagcct ttactaaagt gtggcgtaaa tcacagtagt agcaaagaaa gtgactctgg 4080

atgtgttcct ggccagtacc tcccttatca tgaatgtaga ctctctcatc aagatttagg 4140

aatataaatc aaatcaaatg tgcccagcca agctatgtag taagggactt gaacaatatt 4200

aggcagaacc tataaaataa atcagggaat tagaaattat ttaaagtttt caaattgtaa 4260

attgccccgg tgtctttcag cctactgcca ttatttttgc tacaatacct acatttcaga 4320

ggagggccta ctgaaaattc catgcaagtg gaaaataatc ctcaagttat taatgagttt 4380

gaaaagcaat gagttcttaa gtctttgtga gtagagcaag atcctacaaa attcagaaat 4440

agtaaaaatg gattcatgct gatttgaaga gcatctgtgt gcataatata atgctgcatc 4500

tcttttaaaa gcagtctatt tttcttttta aatttgtccc catagatgct tttgaacatg 4560

aacatgctta tgttaccttt tccgaggttg ggaagagcca ggagctctca ggcagggccc 4620

cctccctcag ctgggcagga gctgctcagg aggagctagt tatagaggaa gcttagcgtt 4680

ggcattttca aaattcaagg tgataacgct ttcttcttcc tttctgtttt agaatagatt 4740

gctgtctgat ttgaaaaagg gaaatagatt tgatctcaaa tgaatctgtg cccagaagcc 4800

aggctcaggg tattcagaga tttgtatagt gccctcaaaa aataacaaaa ttttagcttt 4860

ccttttttct tcttttctcc atcaaattct tttttctcta gtttacaaat gacatggaaa 4920

aggaatttcc cctgagtttt gtatgccttt ttttttttgg cttagactat agataggcgt 4980

gttgagctcc taagaaaata caaggaggaa ctctttgttg tgcagagcac tttatgagta 5040

gtttgtgtgg ataatatgtg actgcttccc tgacgagctt gtgaggctgt acttatgtct 5100

ttcctgtaag gcagcttcag tgccttctgt agtgtatata aggaaagatt acgccttctg 5160

aaaaatctca gagcaaccat aagattattt taaaatatgt agtatgactg atggactttt 5220

tcatcattaa attagtctag catctaaact tttaccactg aaataatatt gaccaaaaag 5280

caatttataa aaggtatttg tgaatagaaa atacaatgtg atcatttgta cttatgtgca 5340

ccttaaaaga ggaattctgt ctagctgtca aattctggtt ccttaacatc cagtccttga 5400

ttgtgattga gatctggtag gacgtgctgg ggcacgctag cagataaaat cccgtatact 5460

ttaggataga tgttacattt atgtcagtgt tggcaaagag cattgtgtag taataaagaa 5520

ttcaagactt cagcaatgtc aacctgaaac tttgtaaata tttcctagat tgttatttga 5580

tgcagtcaca gctctttatc acacaatgtt gtctttccct catcaggcaa ttttagaact 5640

gctgcacacc cctcctcaga tctcacctgc ccctcctgta cattcacctc tccagccttg 5700

tgcacacctc atttagcttt agtttgaaac acattgcagg gttcaggtga cctcttcaaa 5760

aactacctcc tcagaatgag gtaatgaata gttatttatt ttaaaatatg aaaagtcagg 5820

agctctagaa tatgaagatg atctaagatt ttaactttta tgtatacttg ttgagcactc 5880

tccttttgtc ctaaagggca ttatacattt aagcagtaat actgaaaaat gtagctcaga 5940

gtaactgaat gttgttgaaa gtggtgccag aatctgtttt aggggtacgt atcagaatct 6000

taatcttaaa tcggttacat gaaattaaat agttaatggt aacacttgac taacagatat 6060

aattttaatt ttcggtaggc ttttagcaag acagtaagta catcttcata atgagttagc 6120

cacagcttca tcacatgcac agattttcct gttgagagac tgcccagtta agagggtaga 6180

atgatgaacc atttttcagg attctcttct ttgtccaaac tggcattgtg agtgctagaa 6240

tatcagcact ttcaaactag tgattccaac tattaggcta ttaaaaagca aaacaaacca 6300

aacaaaccat agccagacat gggaagttta ctatgagtat aaacagcaaa tagcttacag 6360

gtcatacatt gaaatggtgt aggtaaggcg ttagaaaaat accttgacaa tttgccaaat 6420

gatcttactg tgccttcatg atgcaataaa aaaaaaaaaa atttagcata aatcagtgat 6480

ttgtgaagag agcagccacc ctggtctaac tcagctgtgt taatattttt tagcgtgcaa 6540

tttagactgc aaagataaat gcactaaaga gtttatagcc aaaatcacat ttaaaaaatg 6600

agagaaaaca caggtaaatt ttcagtgaac aaaattattt ttttaaagta cataatccct 6660

agtatagtca gatatattta tcacatagag caaataggtt gaaatcacaa ttcagtgaca 6720

tttctagaga aactttttct actcccatag gttcttcaaa gcatggaact tttatataac 6780

agaaatgtgt gacggtcatt ttaaattgct gtagtttggg gctgaagtac tgtgtgctgg 6840

gcagcaatca catgtattaa ctagtgagaa aggagaaatt aagatatagg acagaatttg 6900

attttcttgt tcccagatta ctgctgccaa cctagacact gagtttccag aggctgaaac 6960

gtaaacttgc agctcagcaa ctgttttgca aagttagtgg gactgtcctg cttatgctgt 7020

tcaaaaatgc tctgagggcc aggtggggcc tccaggggct cctctctgag gggacatcag 7080

actagctaac gacctggcgg gcggatgtga accggacaca ctccatggtg tgcttcttgt 7140

atcggtccct cgccaccctc aagaaaggct tcagcgggtt ctctagacgt ctccactaag 7200

gtgtgttact aacagccatg ggttgttgag cacccgagga gtgcaatagc atctctgcat 7260

gattgtatat tggcccgaag agaatgaagt ggccagtgta ctcatgttcc atgttgctag 7320

ctctggtaaa ctgaaaatac tggtaagatt tttgttttat cagtacacta gagagtaagc 7380

tttgttttgt tgtttttaga taatgttttc acttccattt ggaaagacat ttaaattgag 7440

tttcagtcct aaattttgcc agtcatggta attagcagtt tctatcaggt atttttaagg 7500

tagaagagga tagaaacata agttctaaaa gcttaaggta accgtggttt attttaaaat 7560

gtttaggggt ggttagtctc tacctcaaaa aaagtgagtg aatcttttat ttcagcattc 7620

acaagttcgg ctgttgtttt tgtaatacat ttttttttta accttttgac ccccctttac 7680

ctaagtgtca atgtagtttt attaattact aagtcagttt cattaaaatg tttatttagc 7740

agttttgact aattgcaatg attaatatag ccagttgtgc atgaggacac agccagtgag 7800

tatatctggg ttttttttgt gatgcttttt ttcttaagac ttctgtagat ttatgaagta 7860

ctcattgaaa acaactaaaa tacgtttatt cgtgttaata tggaaaaaaa aaaa 7914

<210>24

<211>766

<212>PRT

<213> cattle

<400>24

Met Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu

1 5 10 15

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

130 135 140

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

275 280 285

Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala

290 295 300

Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro

305 310 315 320

Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser

325 330 335

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

340 345 350

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

355 360 365

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp

370 375 380

Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr

385 390 395 400

Pro ProAla Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln

405 410 415

Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu

420 425 430

Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp

435 440 445

Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser

450 455 460

Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala

465 470 475 480

Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala

485 490 495

Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile

500 505 510

Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr

515 520 525

Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu

530 535 540

Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala

545 550 555 560

Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu

565 570 575

Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly

580 585 590

Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln

595 600 605

Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile

610 615 620

Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala

625 630 635 640

Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser

645 650 655

Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr

660 665 670

Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met

675 680 685

Lys Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro

690 695 700

Leu Phe Pro Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu

705 710 715 720

Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala Gly

725 730 735

Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys Thr

740 745 750

Pro Ile Gln Ala Gly Gly Tyr Gly Glu Phe Ala Ala Phe Lys

755 760 765

<210>25

<211>4670

<212>DNA

<213> cattle

<400>25

ggtgtgtcat agtgcagcag attgaatgca gaagatatga aaattcagat gtcttctgtt 60

aaggtgtgga atatcaaaca aatgattaag ttgacacagg agcatataga ggccctattg 120

gacaaatttg gtggggagca taatccacca tcaatatatc tggaggccta tgaagaatac 180

accagcaagc tagatgccct ccaacaaagagaacaacagt tattggaatc cctggggaat 240

ggaactgatt tttctgtttc tagctctgca tcaacggaca ccgttacatc ttcttcctct 300

tctagccttt cagtgctgcc ttcatctctt tcagtttttc aaaatcccac agatgtgtca 360

cggagcaacc ccaagtcacc acaaaaacct atcgttagag tcttcctgcc caataaacag 420

aggacagtgg tacctgcacg gtgtggagtc acagtccggg acagcctgaa gaaggcactg 480

atgatgagag gtctaatccc agagtgctgt gctgtttaca gaattcagga tggggagaag 540

aaaccaattg gctgggacac tgatatttcc tggcttactg gagaggagtt gcatgtagaa 600

gtgttggaga atgttccact tacaacacac aactttgtac ggaaaacttt tttcacctta 660

gcattttgtg acttctgtag aaagctgctt ttccagggat tccgctgtca aacatgtggt 720

tataaatttc accagcgttg tagtacagag gttccactga tgtgtgttaa ttatgaccaa 780

ctagatttgc tgtttgtctc caagttcttt gaacaccacc caataccaca ggaggaggcc 840

tccttagcag agactaccct tccatgtggc tcatcccctt ctgcaccccc ctccgattct 900

attgggcccc caattctcac cagtccatct ccttcaaaat ccattccaat tccacagcct 960

ttccgaccag cagatgaaga tcatcgaaat cagtttggac aacgagaccg gtcctcatca 1020

gctccaaatg tgcatataaa cacaatagaa cccgtcaata ttgatgactt gattagagac 1080

caagggtttc gtagtgatgg aggatcaacc acaggtttat ccgccacacc ccctgcctca 1140

ttacctggct cactctctaa tgtgaaagca ttgcagaaat ctccaggacc tcagcgagaa 1200

agaaagtcct cttcatcctc agaagacagg aatcgaatga aaacgcttgg tagacgggat 1260

tcaagtgacg attgggagat tcctgatgga cagatcacag tgggacaaag aattggatca 1320

gggtcatttg ggacagtcta caagggaaag tggcatggtg atgtggcagt gaaaatgttg 1380

aatgtgacag cacccacacc tcagcagtta caggccttca aaaatgaagt aggagtactc 1440

aggaaaacgc gacatgtgaa tatcctcctc ttcatgggtt attcaacaaa gccacaactg 1500

gctattgtta cccagtggtg tgagggctcc agtttatatc atcatctcca catcattgag 1560

accaaattcg agatgatcaa acttatagat attgcacggc agactgcaca gggcatggat 1620

tacttacacg ccaagtcaat catccacaga gacctcaaga gtaataatat ttttcttcat 1680

gaagacctca cagtaaaaat aggtgatttt ggtctagcca cagtgaaatc tcgatggagt 1740

gggtcccatc agtttgaaca gttgtctgga tccattttgt ggatggcacc agaagtaatc 1800

agaatgcaag ataaaaaccc atatagcttt cagtcagatg tatatgcatt tgggattgtt 1860

ctgtatgaat tgatgaccgg acagttacct tattcaaata tcaacaacag ggaccagata 1920

atttttatgg tgggacgagg atatctgtct ccagatctca gtaaggtacg gagtaactgt 1980

ccaaaagcca tgaagagatt aatggcagag tgcctaaaaa agaaaagaga tgaaagacca 2040

ctctttcccc aagtaggaaa gactctccta agcaagagac aaaattcaga agttatcagg 2100

gaaaaagata agcagattct cgcctctatt gagctgctgg cccgctcatt gccaaaaatt 2160

caccgcagtg catcagaacc ctccttgaat cgggctggct tccaaacaga ggattttagt 2220

ctatatgctt gtgcttctcc aaaaacaccc attcaggcag ggggatatga agcagatttg 2280

gctcttacat caaataaaaa tagagtagaa gttgggattt agagatttcc tgacatgcaa 2340

gaaggaataa gcaagaaaaa aaggtttgtt ttccccaaat catatctatt gtcttttact 2400

tctatttttt cttaaatttt ttgtgatttc agagacatgt agagttttat tgatacctaa 2460

actatgagtt cttttttttt tttttttttc attattttga tttttttggc caagaggcat 2520

atgggatctt agcttgagaa agcaacaatt ttcttgatgt cattttgggt gagggcacat 2580

attgctgtga acagtgtggt gatagccacc agggaccaaa ctcacacccg ctgcattgaa 2640

aggtgaagtc ttaaacactg gaccagcaga gaaattccta ctctatgagt tctttttgtc 2700

atcccctccc cgcaccctcc acccccaacc taaagtctga tgatgaaatc aacaactatt 2760

ccattagaag cagtagattc tggtagcatg atctttagtt tgttagtaag attttgtgct 2820

ttgtggggtt gtgtcgtttt aaggctaata tttaagtttg tcaaatagaa tgctgttcag 2880

attgtaaaaa tgagtaataa acatctgaag ttttttttaa gttattttta acatggtata 2940

tacagttgag cttagagttt atcattttct gatattctct tacttagtag atgaattcta 3000

gccatttttt ataaagattt ctgttaagca aatcctgttt tcacatgggc ttcctttaag 3060

ggattttaga ttctgctgga tatggtgact gctcataaga ctgttgaaaa ttacttttaa 3120

gatgtattag aatacttctg aaaaaaaata gcaaccttaa aaccataagc aaaagtagta 3180

agggtgttta tacatttcta gagtccctgt ttaggtaata gcctcctatg attgtacttt 3240

aaatgttttg ctctccaagg ttttagtaac ttggcttttt ttctaatcag tgccaaactc 3300

ccccagtttt tttaacttta aatatgaggt aataaatctt ttacccttcc ttgatctttt 3360

gacttataat accttggtca gttgtttctt aaaaggaatc cttaaatgga aagagacaat 3420

atcactgtct gcagttctga ttagtagttt tattcagaat ggaaaaacag attattcatt 3480

tttgaaaatt gttcaggggt atgttcattg ttaggacctt ggactttgga gtcagtgcct 3540

agctatgcat tccaggtctg ccattttctg gctgtgaaat tttggacaag ttacttaacc 3600

actttaaacc ccagctttaa gaagtaaatt aaccccagta aattaagaag taatagcagc 3660

cacttcgtag agttgttatg aggctcagat gcagtgcaaa tgtgtataaa gtattcaggg 3720

agtcacctgg tatactataa tagacactag aatagttgcc aatattatca gcatacaatc 3780

tgaggattct gtcagccaat cattagcaat ctgttgtttg ttgggacatg ccagtgttct 3840

ccagttgaaa tcagtagcaa tctaaaaatg gatagattat tcctcattta aatagtgtgt 3900

tcatataagt gattgcttgg atccttatca gaagttgctg ttactgaaaa atgataaggc 3960

tgactaaatt gtgatagttg tcagttacta accaactccc agaaatgaat aagaggaacc 4020

tatctctagt tcctagtaga aggtatggac aaaatagtag gtgaaaaata atgtcttgaa 4080

cccccaaatt aagtaagctt taaagagtac aatacctcaa agggtctttg cggtttaaaa 4140

tttgtatgct gagaatgatg ttcattgaca tgtgcctata tgtaattttt tgatagttta 4200

aaaggtgaaa tgaactacag atgggagagg tctgaatttt cttgccttca gtcaaatgtg 4260

taatgtggac atattatttg acctgtgaat tttatctttt aaaaaagatt aattcctgct 4320

tcttccttcc taatagttgc attataataa tgaaaatgag ttgataattt ggggggaaag 4380

tattctacaa atcaacctta ttattttacc attggtttct gagaaatttt gttcatttga 4440

accgtttata gcttgattag aatcatagca tgtaaaaccc aactgaggga ttatctgcag 4500

acttaatgta gtattatgta agttgtcttc tttcatttcg accttttttg cttttgttgt 4560

tgctagatct gtagtatgta gctagtcacc tttcagcgag gtttcagcga ggcttttctg 4620

tgtctctagg ttatttgaga taactttttt aaaattagct cttgtcctcc 4670

<210>26

<211>761

<212>PRT

<213> cattle

<400>26

Met Lys Ile Gln Met Ser Ser Val Lys Val Trp Asn Ile Lys Gln Met

1 5 10 15

Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu Asp Lys Phe Gly

20 25 30

Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala Tyr Glu Glu Tyr

35 40 45

Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln Gln Leu Leu Glu

50 55 60

Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser Ser Ala Ser Thr

65 70 75 80

Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser Val Leu Pro Ser

85 90 95

Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser Arg Ser Asn Pro

100 105 110

Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu Pro Asn Lys Gln

115 120 125

Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val Arg Asp Ser Leu

130 135 140

Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys Cys Ala Val

145 150 155 160

Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile GlyTrp Asp Thr Asp

165 170 175

Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu Val Leu Glu Asn

180 185 190

Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr Phe Phe Thr Leu

195 200 205

Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln Gly Phe Arg Cys

210 215 220

Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser Thr Glu Val Pro

225 230 235 240

Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu Phe Val Ser Lys

245 250 255

Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala Ser Leu Ala Glu

260 265 270

Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro Pro Ser Asp Ser

275 280 285

Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser Lys Ser Ile Pro

290 295 300

Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His Arg Asn Gln Phe

305 310 315 320

Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val His Ile Asn Thr

325 330 335

Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp Gln Gly Phe Arg

340 345 350

Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr Pro Pro Ala Ser

355 360 365

Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln Lys Ser Pro Gly

370 375 380

Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn Arg

385 390 395 400

Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro

405 410 415

Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly

420 425 430

Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala Val Lys Met Leu

435 440445

Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu

450 455 460

Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met

465 470 475 480

Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu

485 490 495

Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu Thr Lys Phe Glu

500 505 510

Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met Asp

515 520 525

Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn Asn

530 535 540

Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly Asp Phe Gly Leu

545 550 555 560

Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu

565 570 575

Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp

580 585 590

Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val

595 600 605

Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn

610 615 620

Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr Leu Ser Pro Asp

625 630 635 640

Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met Lys Arg Leu Met

645 650 655

Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln

660 665 670

Val Gly Lys Thr Leu Leu Ser Lys Arg Gln Asn Ser Glu Val Ile Arg

675 680 685

Glu Lys Asp Lys Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser

690 695 700

Leu Pro Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala

705 710 715 720

Gly Phe Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys

725 730 735

Thr Pro Ile Gln Ala Gly Gly Tyr Glu Ala Asp Leu Ala Leu Thr Ser

740 745 750

Asn Lys Asn Arg Val Glu Val Gly Ile

755 760

<210>27

<211>4816

<212>DNA

<213> cattle

<400>27

ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60

cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120

ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180

ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240

agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt 300

cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg attaagttga 360

cacaggagca tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa 420

tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac 480

aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540

cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag 600

tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660

ttagagtctt cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720

tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg 780

tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc 840

ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact 900

ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960

agggattccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc 1020

cactgatgtg tgttaattat gaccaactag agcccccaat tctcaccagt ccatctcctt 1080

caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1140

ttggacaacg agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg 1200

tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag 1260

gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc 1320

agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc 1380

gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct gatggacaga 1440

tcacagtggg acaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc 1500

atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag cagttacagg 1560

ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1620

tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt 1680

tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt atagatattg 1740

cacggcagac tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc 1800

tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc 1860

tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca 1920

ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt 1980

cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag ttaccttatt 2040

caaatatcaa caacagggac cagataattt ttatggtggg acgaggatat ctgtctccag 2100

atctcagtaa ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc 2160

taaaaaagaa aagagatgaa agaccactct ttccccaagt aggaaagact ctcctaagca 2220

agagacaaaa ttcagaagtt atcagggaaa aagataagca gattctcgcc tctattgagc 2280

tgctggcccg ctcattgcca aaaattcacc gcagtgcatc agaaccctcc ttgaatcggg 2340

ctggcttcca aacagaggat tttagtctat atgcttgtgc ttctccaaaa acacccattc 2400

aggcaggggg atatgaagca gatttggctc ttacatcaaa taaaaataga gtagaagttg 2460

ggatttagag atttcctgac atgcaagaag gaataagcaa gaaaaaaagg tttgttttcc 2520

ccaaatcata tctattgtct tttacttcta ttttttctta aattttttgt gatttcagag 2580

acatgtagag ttttattgat acctaaacta tgagttcttt tttttttttt tttttcatta 2640

ttttgatttt tttggccaag aggcatatgg gatcttagct tgagaaagca acaattttct 2700

tgatgtcatt ttgggtgagg gcacatattg ctgtgaacag tgtggtgata gccaccaggg 2760

accaaactca cacccgctgc attgaaaggt gaagtcttaa acactggacc agcagagaaa 2820

ttcctactct atgagttctt tttgtcatcc cctccccgca ccctccaccc ccaacctaaa 2880

gtctgatgat gaaatcaaca actattccat tagaagcagt agattctggt agcatgatct 2940

ttagtttgtt agtaagattt tgtgctttgt ggggttgtgt cgttttaagg ctaatattta 3000

agtttgtcaa atagaatgct gttcagattg taaaaatgag taataaacat ctgaagtttt 3060

ttttaagtta tttttaacat ggtatataca gttgagctta gagtttatca ttttctgata 3120

ttctcttact tagtagatga attctagcca ttttttataa agatttctgt taagcaaatc 3180

ctgttttcac atgggcttcc tttaagggat tttagattct gctggatatg gtgactgctc 3240

ataagactgt tgaaaattac ttttaagatg tattagaata cttctgaaaa aaaatagcaa 3300

ccttaaaacc ataagcaaaa gtagtaaggg tgtttataca tttctagagt ccctgtttag 3360

gtaatagcct cctatgattg tactttaaat gttttgctct ccaaggtttt agtaacttgg 3420

ctttttttct aatcagtgcc aaactccccc agttttttta actttaaata tgaggtaata 3480

aatcttttac ccttccttga tcttttgact tataatacct tggtcagttg tttcttaaaa 3540

ggaatcctta aatggaaaga gacaatatca ctgtctgcag ttctgattag tagttttatt 3600

cagaatggaa aaacagatta ttcatttttg aaaattgttc aggggtatgt tcattgttag 3660

gaccttggac tttggagtca gtgcctagct atgcattcca ggtctgccat tttctggctg 3720

tgaaattttg gacaagttac ttaaccactt taaaccccag ctttaagaag taaattaacc 3780

ccagtaaatt aagaagtaat agcagccact tcgtagagtt gttatgaggc tcagatgcag 3840

tgcaaatgtg tataaagtat tcagggagtc acctggtata ctataataga cactagaata 3900

gttgccaata ttatcagcat acaatctgag gattctgtca gccaatcatt agcaatctgt 3960

tgtttgttgg gacatgccag tgttctccag ttgaaatcag tagcaatcta aaaatggata 4020

gattattcct catttaaata gtgtgttcat ataagtgatt gcttggatcc ttatcagaag 4080

ttgctgttac tgaaaaatga taaggctgac taaattgtga tagttgtcag ttactaacca 4140

actcccagaa atgaataaga ggaacctatc tctagttcct agtagaaggt atggacaaaa 4200

tagtaggtga aaaataatgt cttgaacccc caaattaagt aagctttaaa gagtacaata 4260

cctcaaaggg tctttgcggt ttaaaatttg tatgctgaga atgatgttca ttgacatgtg 4320

cctatatgta attttttgat agtttaaaag gtgaaatgaa ctacagatgg gagaggtctg 4380

aattttcttg ccttcagtca aatgtgtaat gtggacatat tatttgacct gtgaatttta 4440

tcttttaaaa aagattaatt cctgcttctt ccttcctaat agttgcatta taataatgaa 4500

aatgagttga taatttgggg ggaaagtatt ctacaaatca accttattat tttaccattg 4560

gtttctgaga aattttgttc atttgaaccg tttatagctt gattagaatc atagcatgta 4620

aaacccaact gagggattat ctgcagactt aatgtagtat tatgtaagtt gtcttctttc 4680

atttcgacct tttttgcttt tgttgttgct agatctgtag tatgtagcta gtcacctttc 4740

agcgaggttt cagcgaggct tttctgtgtc tctaggttat ttgagataac ttttttaaaa 4800

ttagctcttg tcctcc 4816

<210>28

<211>757

<212>PRT

<213> cattle

<400>28

Met Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu

1 5 10 15

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

130 135 140

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Glu Pro Pro

275 280 285

Ile Leu Thr Ser Pro Ser Pro Ser Lys Ser Ile Pro Ile Pro Gln Pro

290 295 300

Phe Arg Pro Ala Asp Glu Asp His Arg Asn Gln Phe Gly Gln Arg Asp

305 310 315 320

Arg Ser Ser Ser Ala Pro Asn Val His Ile Asn Thr Ile Glu Pro Val

325 330 335

Asn Ile Asp Asp Leu Ile Arg Asp Gln Gly Phe Arg Ser Asp Gly Gly

340 345 350

Ser Thr Thr Gly Leu Ser Ala Thr Pro Pro Ala Ser Leu Pro Gly Ser

355 360 365

Leu Ser Asn Val Lys Ala Leu Gln Lys Ser Pro Gly Pro Gln Arg Glu

370 375 380

Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn Arg Met Lys Thr Leu

385 390 395 400

Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly Gln Ile

405 410 415

Thr Val Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr Val Tyr Lys

420 425 430

Gly Lys Trp His Gly Asp Val Ala Val Lys Met Leu Asn Val Thr Ala

435 440 445

Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val Gly Val Leu

450 455 460

Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met Gly Tyr Ser Thr

465 470 475 480

Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu Gly Ser Ser Leu

485 490 495

Tyr His His Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys Leu

500 505 510

Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His Ala

515 520 525

Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe Leu His

530 535 540

Glu Asp Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr Val Lys

545 550 555 560

Ser Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser Gly Ser Ile

565 570 575

Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys Asn Pro Tyr

580 585 590

Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu Tyr Glu Leu

595 600 605

Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln Ile

610 615 620

Ile Phe Met Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys Val

625 630 635 640

Arg Ser Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys Leu

645 650 655

Lys Lys Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Val Gly Lys Thr

660 665 670

Leu Leu Ser Lys Arg Gln Asn Ser Glu Val Ile Arg Glu Lys Asp Lys

675 680 685

Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro Lys Ile

690 695 700

His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe Gln Thr

705 710 715 720

Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys Thr Pro Ile Gln

725 730 735

Ala Gly Gly Tyr Glu Ala Asp Leu Ala Leu Thr Ser Asn Lys Asn Arg

740 745 750

Val Glu Val Gly Ile

755

<210>29

<211>2499

<212>DNA

<213> cattle

<400>29

ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60

cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120

ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180

ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240

agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt 300

cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg attaagttga 360

cacaggagca tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa 420

tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac 480

aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540

cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag 600

tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660

ttagagtctt cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720

tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg 780

tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc 840

ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact 900

ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960

agggattccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc 1020

cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080

accacccaat accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat 1140

ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt 1200

caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260

ttggacaacg agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg 1320

tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380

gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440

agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc 1500

gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct gatggacaga 1560

tcacagtggg acaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc 1620

atggtgatgt ggcagtgaaa atgttgaatg tgacagcacccacacctcag cagttacagg 1680

ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740

tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt 1800

tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt atagatattg 1860

cacggcagac tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc 1920

tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc 1980

tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040

ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt 2100

cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag ttaccttatt 2160

caaatatcaa caacagggac cagataattt ttatggtggg acgaggatat ctgtctccag 2220

atctcagtaa ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc 2280

taaaaaagaa aagagatgaa agaccactct ttccccaagt aggaaagact ctcctaagca 2340

agagacaaaa ttcagaagtt atcagggaaa aagataagca ggaaaagtat gtttctttag 2400

tacattccag gcatttggga ttacagtaaa aacaatattc tcgcctctat tgagctgctg 2460

gcccgctcat tgccaaaaat tcaccgcagt gcatcagaa 2499

<210>30

<211>744

<212>PRT

<213> cattle

<400>30

Met Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu

1 5 10 15

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu ProSer Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

130 135 140

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

275 280 285

Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala

290 295 300

Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro

305 310 315 320

Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser

325 330 335

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

340 345 350

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

355 360 365

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp

370 375 380

Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr

385 390 395 400

Pro Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln

405 410 415

Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu

420 425 430

Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp

435 440 445

Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser

450 455 460

Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala

465 470 475 480

Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala

485 490 495

Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile

500 505 510

Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr

515 520 525

Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu

530 535 540

Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala

545 550 555 560

Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu

565 570 575

Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly

580 585 590

Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln

595 600 605

Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile

610 615 620

Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala

625 630 635 640

Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser

645 650 655

Asn Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr

660 665 670

Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met

675 680 685

Lys ArgLeu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro

690 695 700

Leu Phe Pro Gln Val Gly Lys Thr Leu Leu Ser Lys Arg Gln Asn Ser

705 710 715 720

Glu Val Ile Arg Glu Lys Asp Lys Gln Glu Lys Tyr Val Ser Leu Val

725 730 735

His Ser Arg His Leu Gly Leu Gln

740

<210>31

<211>2404

<212>DNA

<213> cattle

<400>31

ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60

cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120

ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180

ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240

agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt 300

cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg attaagttga 360

cacaggagca tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa 420

tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac 480

aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540

cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag 600

tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660

ttagagtctt cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720

tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg 780

tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc 840

ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact 900

ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960

agggattccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc 1020

cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080

accacccaat accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat 1140

ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt 1200

caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260

ttggacaacg agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg 1320

tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380

gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440

agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc 1500

gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct gatggacaga 1560

tcacagtggg acaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc 1620

atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag cagttacagg 1680

ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740

tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt 1800

tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt atagatattg 1860

cacggcagac tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc 1920

tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc 1980

tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040

ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt 2100

cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag ttaccttatt 2160

caaatatcaa caacagggac cagataattt ttatggtggg acgaggatat ctgtctccag 2220

atctcagtaa ggtacggagt aactgtccaa aagccatgaa gagattaatg gcagagtgcc 2280

taaaaaagaa aagagatgaa agaccactct ttccccaaga tctctcttcc caccatagac 2340

acaaaaattt cagatggcta caggtttaca tgtaaaaaac agaattataa caaatgattt 2400

ttat 2404

<210>32

<211>726

<212>PRT

<213> cattle

<400>32

Met Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu

1 5 10 15

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly AsnGly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

130 135 140

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

275 280 285

Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala

290 295 300

Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro

305 310 315 320

Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser

325 330 335

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

340 345 350

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

355 360 365

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp

370 375380

Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr

385 390 395 400

Pro Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln

405 410 415

Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu

420 425 430

Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp

435 440 445

Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser

450 455 460

Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala

465 470 475 480

Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala

485 490 495

Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile

500 505 510

Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr

515 520 525

Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu

530 535 540

Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala

545 550 555 560

Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu

565 570 575

Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly

580 585 590

Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln

595 600 605

Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile

610 615 620

Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala

625 630 635 640

Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser

645 650 655

Asn Ile Asn Asn Arg Asp GlnIle Ile Phe Met Val Gly Arg Gly Tyr

660 665 670

Leu Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met

675 680 685

Lys Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro

690 695 700

Leu Phe Pro Gln Asp Leu Ser Ser His His Arg His Lys Asn Phe Arg

705 710 715 720

Trp Leu Gln Val Tyr Met

725

<210>33

<211>2331

<212>DNA

<213> cattle

<400>33

ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60

cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120

ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180

ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240

agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt 300

cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg attaagttga 360

cacaggagca tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa 420

tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac 480

aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540

cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag 600

tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660

ttagagtctt cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720

tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg 780

tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc 840

ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact 900

ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960

agggattccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc 1020

cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080

accacccaat accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat 1140

ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt 1200

caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260

ttggacaacg agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg 1320

tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380

gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440

agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc 1500

gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct gatggacaga 1560

tcacagtggg acaaagaatt ggatcagggt catttgggac agtctacaag ggaaagtggc 1620

atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag cagttacagg 1680

ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740

tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt 1800

tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt atagatattg 1860

cacggcagac tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc 1920

tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc 1980

tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040

ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt 2100

cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag ttaccttatt 2160

caaatatcaa caacagggac caggtgcttt gtcctccatg ggagtgtaat aaatgctgtg 2220

caagggctta cttcccatga gagaagtgag tgaccaacag aaggataatt tttatggtgg 2280

gacgaggata tctgtctcca gatctcagta aggtacggag taactgtcca a 2331

<210>34

<211>681

<212>PRT

<213> cattle

<400>34

Met Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu

1 5 10 15

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

130 135 140

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

275 280 285

Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala

290 295 300

Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro

305 310 315 320

Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser

325 330 335

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

340 345 350

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

355 360 365

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp

370 375380

Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr

385 390 395 400

Pro Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln

405 410 415

Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu

420 425 430

Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp

435 440 445

Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser

450 455 460

Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala

465 470 475 480

Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala

485 490 495

Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile

500 505 510

Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr

515 520 525

Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu

530 535 540

Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala

545 550 555 560

Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu

565 570 575

Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly

580 585 590

Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln

595 600 605

Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile

610 615 620

Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala

625 630 635 640

Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser

645 650 655

Asn Ile Asn Asn Arg Asp GlnVal Leu Cys Pro Pro Trp Glu Cys Asn

660 665 670

Lys Cys Cys Ala Arg Ala Tyr Phe Pro

675 680

<210>35

<211>2319

<212>DNA

<213> cattle

<400>35

ctcagctgcg ccgggtctca caagacggtt cccgaggtgg cccaggcgcc gtcccaccgc 60

cgacgccgcc cgggccgccc gggccgtccc tccccgctgc cccccgtcct ccgcctccgc 120

ctccccccgc cctcagcctc ccttccccct ccccgcccag cagcggtcgc tcgggcccgg 180

ctctcggtta taagatggcg gcgctgagtg gcggcggcgg cggcggcggc ggtggcgcgg 240

agcagggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc gcggcctctt 300

cggctgcgga ccccgccatt cccgaggagg tgtggaatat caaacaaatg attaagttga 360

cacaggagca tatagaggcc ctattggaca aatttggtgg ggagcataat ccaccatcaa 420

tatatctgga ggcctatgaa gaatacacca gcaagctaga tgccctccaa caaagagaac 480

aacagttatt ggaatccctg gggaatggaa ctgatttttc tgtttctagc tctgcatcaa 540

cggacaccgt tacatcttct tcctcttcta gcctttcagt gctgccttca tctctttcag 600

tttttcaaaa tcccacagat gtgtcacgga gcaaccccaa gtcaccacaa aaacctatcg 660

ttagagtctt cctgcccaat aaacagagga cagtggtacc tgcacggtgt ggagtcacag 720

tccgggacag cctgaagaag gcactgatga tgagaggtct aatcccagag tgctgtgctg 780

tttacagaat tcaggatggg gagaagaaac caattggctg ggacactgat atttcctggc 840

ttactggaga ggagttgcat gtagaagtgt tggagaatgt tccacttaca acacacaact 900

ttgtacggaa aacttttttc accttagcat tttgtgactt ctgtagaaag ctgcttttcc 960

agggattccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt acagaggttc 1020

cactgatgtg tgttaattat gaccaactag atttgctgtt tgtctccaag ttctttgaac 1080

accacccaat accacaggag gaggcctcct tagcagagac tacccttcca tgtggctcat 1140

ccccttctgc acccccctcc gattctattg ggcccccaat tctcaccagt ccatctcctt 1200

caaaatccat tccaattcca cagcctttcc gaccagcaga tgaagatcat cgaaatcagt 1260

ttggacaacg agaccggtcc tcatcagctc caaatgtgca tataaacaca atagaacccg 1320

tcaatattga tgacttgatt agagaccaag ggtttcgtag tgatggagga tcaaccacag 1380

gtttatccgc cacaccccct gcctcattac ctggctcact ctctaatgtg aaagcattgc 1440

agaaatctcc aggacctcag cgagaaagaa agtcctcttc atcctcagaa gacaggaatc 1500

gaatgaaaac gcttggtaga cgggattcaa gtgacgattg ggagattcct gatggacaga 1560

tcacagtggg acaaagaatt ggatcagggtcatttgggac agtctacaag ggaaagtggc 1620

atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc cacacctcag cagttacagg 1680

ccttcaaaaa tgaagtagga gtactcagga aaacgcgaca tgtgaatatc ctcctcttca 1740

tgggttattc aacaaagcca caactggcta ttgttaccca gtggtgtgag ggctccagtt 1800

tatatcatca tctccacatc attgagacca aattcgagat gatcaaactt atagatattg 1860

cacggcagac tgcacagggc atggattact tacacgccaa gtcaatcatc cacagagacc 1920

tcaagagtaa taatattttt cttcatgaag acctcacagt aaaaataggt gattttggtc 1980

tagccacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg tctggatcca 2040

ttttgtggat ggcaccagaa gtaatcagaa tgcaagataa aaacccatat agctttcagt 2100

cagatgtata tgcatttggg attgttctgt atgaattgat gaccggacag ttaccttatt 2160

caaatatcaa caacagggac caggtgcttt gtcctccatg ggagtgtaat aaatgctgtg 2220

caagggctta cttcccatga gagaagtgag tgaccaacag aaggtctgtg caaggaaaag 2280

agacaaagcc acggatcaga agcacatggc cataactga 2319

<210>36

<211>681

<212>PRT

<213> cattle

<400>36

Met Ala Ala Leu Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ala Glu

1 5 10 15

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

130 135 140

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

275 280 285

Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala

290 295 300

Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro

305 310 315 320

Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser

325 330 335

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

340 345 350

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

355 360 365

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp

370 375 380

Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr

385 390 395 400

Pro Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln

405 410 415

Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu

420425 430

Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp

435 440 445

Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser

450 455 460

Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala

465 470 475 480

Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala

485 490 495

Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile

500 505 510

Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr

515 520 525

Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu

530 535 540

Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala

545 550 555 560

Gln Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu

565 570 575

Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly

580 585 590

Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln

595 600 605

Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile

610 615 620

Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala

625 630 635 640

Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser

645 650 655

Asn Ile Asn Asn Arg Asp Gln Val Leu Cys Pro Pro Trp Glu Cys Asn

660 665 670

Lys Cys Cys Ala Arg Ala Tyr Phe Pro

675 680

<210>37

<211>2661

<212>DNA

<213> cattle

<400>37

tcagctgcgc cgggtctcac aagacggttc ccgaggtggc ccaggcgccg tcccaccgcc 60

gacgccgccc gggccgcccg ggccgtccct ccccgctgcc ccccgtcctc cgcctccgcc 120

tccccccgcc ctcagcctcc cttccccctc cccgcccagc agcggtcgct cgggcccggc 180

tctcggttat aagatggcgg cgctgagtgg cggcggcggc ggcggcggcg gtggcgcgga 240

gcagggccag gctctgttca acggggacat ggagcccgag gccggcgccg cggcctcttc 300

ggctgcggac cccgccattc ccgaggaggt gtggaatatc aaacaaatga ttaagttgac 360

acaggagcat atagaggccc tattggacaa atttggtggg gagcataatc caccatcaat 420

atatctggag gcctatgaag aatacaccag caagctagat gccctccaac aaagagaaca 480

acagttattg gaatccctgg ggaatggaac tgatttttct gtttctagct ctgcatcaac 540

ggacaccgtt acatcttctt cctcttctag cctttcagtg ctgccttcat ctctttcagt 600

ttttcaaaat cccacagatg tgtcacggag caaccccaag tcaccacaaa aacctatcgt 660

tagagtcttc ctgcccaata aacagaggac agtggtacct gcacggtgtg gagtcacagt 720

ccgggacagc ctgaagaagg cactgatgat gagaggtcta atcccagagt gctgtgctgt 780

ttacagaatt caggatgggg agaagaaacc aattggctgg gacactgata tttcctggct 840

tactggagag gagttgcatg tagaagtgtt ggagaatgtt ccacttacaa cacacaactt 900

tgtacggaaa acttttttca ccttagcatt ttgtgacttc tgtagaaagc tgcttttcca 960

gggattccgc tgtcaaacat gtggttataa atttcaccag cgttgtagta cagaggttcc 1020

actgatgtgt gttaattatg accaactaga tttgctgttt gtctccaagt tctttgaaca 1080

ccacccaata ccacaggagg aggcctcctt agcagagact acccttccat gtggctcatc 1140

cccttctgca cccccctccg attctattgg gcccccaatt ctcaccagtc catctccttc 1200

aaaatccatt ccaattccac agcctttccg accagcagat gaagatcatc gaaatcagtt 1260

tggacaacga gaccggtcct catcagctcc aaatgtgcat ataaacacaa tagaacccgt 1320

caatattgat gacttgatta gagaccaagg gtttcgtagt gatggaggat caaccacagg 1380

tttatccgcc acaccccctg cctcattacc tggctcactc tctaatgtga aagcattgca 1440

gaaatctcca ggacctcagc gagaaagaaa gtcctcttca tcctcagaag acaggaatcg 1500

aatgaaaacg cttggtagac gggattcaag tgacgattgg gagattcctg atggacagat 1560

cacagtggga caaagaattg gatcagggtc atttgggaca gtctacaagg gaaagtggca 1620

tggtgatgtg gcagtgaaaa tgttgaatgt gacagcaccc acacctcagc agttacaggc 1680

cttcaaaaat gaagtaggag tactcaggaa aacgcgacat gtgaatatcc tcctcttcat 1740

gggttattca acaaagccac aactggctat tgttacccag tggtgtgagg gctccagttt 1800

atatcatcat ctccacatca ttgagaccaa attcgagatg atcaaactta tagatattgc 1860

acggcagact gcacagggca tggattactt acacgccaag tcaatcatcc acagagacct 1920

caagagtaat aatatttttc ttcatgaaga cctcacagta aaaataggtg attttggtct 1980

agccacagtg aaatctcgat ggagtgggtc ccatcagttt gaacagttgt ctggatccat 2040

tttgtggatg gcaccagaag taatcagaat gcaagataaa aacccatata gctttcagtc 2100

agatgtatat gcatttggga ttgttctgta tgaattgatg accggacagt taccttattc 2160

aaatatcaac aacagggacc agtctgtgca aggaaaagag acaaagccac ggatcagaag 2220

cacatggcca taactgaaga ttttgtgaac tctcacaagg aaaaaatttg ctctttgaac 2280

aataagaagg aactcactaa aatgtaactg agaactgttc aacaggttga aagctgaaag 2340

atgccattgg aactgacaaa atgtttctta aacataaatg atgaaacagt gaaactacat 2400

aatatctcct ctggctgaaa cattcaagaa gtttaaaatg cttaagttaa aaataaaatc 2460

ctagtaaaca atggacttac tgtgcaacat agagaatatc ttacgataac ctgtaatgga 2520

aaagaatctg aaaaagaatg tatataactg aatcactttg ctgtaaacta gaatctgaca 2580

caacactgta aatcactaca cttttctgtt gcatgccaaa gattatttaa taacgtcatt 2640

aaaaaattat tttaataatt a 2661

<210>38

<211>679

<212>PRT

<213> cattle

<400>38

Met Ala Ala Leu Ser GlyGly Gly Gly Gly Gly Gly Gly Gly Ala Glu

1 5 10 15

Gln Gly Gln Ala Leu Phe Asn Gly Asp Met Glu Pro Glu Ala Gly Ala

20 25 30

Ala Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn

35 40 45

Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu

50 55 60

Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala

65 70 75 80

Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln

85 90 95

Gln Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser

100 105 110

Ser Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser

115 120 125

Val Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser

130 135 140

Arg Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu

145 150 155 160

Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val

165 170 175

Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu

180 185 190

Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly

195 200 205

Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu

210 215 220

Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr

225 230 235 240

Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln

245 250 255

Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser

260 265 270

Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu

275280 285

Phe Val Ser Lys Phe Phe Glu His His Pro Ile Pro Gln Glu Glu Ala

290 295 300

Ser Leu Ala Glu Thr Thr Leu Pro Cys Gly Ser Ser Pro Ser Ala Pro

305 310 315 320

Pro Ser Asp Ser Ile Gly Pro Pro Ile Leu Thr Ser Pro Ser Pro Ser

325 330 335

Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His

340 345 350

Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val

355 360 365

His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp

370 375 380

Gln Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr

385 390 395 400

Pro Pro Ala Ser Leu Pro Gly Ser Leu Ser Asn Val Lys Ala Leu Gln

405 410 415

Lys Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu

420 425 430

Asp Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp

435 440 445

Trp Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser

450 455 460

Gly Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala

465 470 475 480

Val Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala

485 490 495

Phe Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile

500 505 510

Leu Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr

515 520 525

Gln Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu

530 535 540

Thr Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala

545 550 555 560

Gln Gly Met Asp Tyr Leu His AlaLys Ser Ile Ile His Arg Asp Leu

565 570 575

Lys Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly

580 585 590

Asp Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln

595 600 605

Phe Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile

610 615 620

Arg Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala

625 630 635 640

Phe Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser

645 650 655

Asn Ile Asn Asn Arg Asp Gln Ser Val Gln Gly Lys Glu Thr Lys Pro

660 665 670

Arg Ile Arg Ser Thr Trp Pro

675

<210>39

<211>7434

<212>DNA

<213> cattle

<400>39

acaccgttac atcttcttcc tcttctagcc tttcagtgct gccttcatct ctttcagttt 60

ttcaaaatcc cacagatgtg tcacggagca accccaagtc accacaaaaa cctatcgtta 120

gagtcttcct gcccaataaa cagaggacag tggtacctgc acggtgtgga gtcacagtcc 180

gggacagcct gaagaaggca ctgatgatga gaggtctaat cccagagtgc tgtgctgttt 240

acagaattca ggatggggag aagaaaccaa ttggctggga cactgatatt tcctggctta 300

ctggagagga gttgcatgta gaagtgttgg agaatgttcc acttacaaca cacaactttg 360

tacggaaaac ttttttcacc ttagcatttt gtgacttctg tagaaagctg cttttccagg 420

gattccgctg tcaaacatgt ggttataaat ttcaccagcg ttgtagtaca gaggttccac 480

tgatgtgtgt taattatgac caactagatt tgctgtttgt ctccaagttc tttgaacacc 540

acccaatacc acaggaggag gcctccttag cagagactac ccttccatgt ggctcatccc 600

cttctgcacc cccctccgat tctattgggc ccccaattct caccagtcca tctccttcaa 660

aatccattcc aattccacag cctttccgac cagcagatga agatcatcga aatcagtttg 720

gacaacgaga ccggtcctca tcagctccaa atgtgcatat aaacacaata gaacccgtca 780

atattgatga cttgattaga gaccaagggt ttcgtagtga tggaggatca accacaggtt 840

tatccgccac accccctgcc tcattacctg gctcactctc taatgtgaaa gcattgcaga 900

aatctccagg acctcagcga gaaagaaagt cctcttcatc ctcagaagac aggaatcgaa 960

tgaaaacgct tggtagacgg gattcaagtg acgattggga gattcctgat ggacagatca 1020

cagtgggaca aagaattgga tcagggtcat ttgggacagt ctacaaggga aagtggcatg 1080

gtgatgtggc agtgaaaatg ttgaatgtga cagcacccac acctcagcag ttacaggcct 1140

tcaaaaatga agtaggagta ctcaggaaaa cgcgacatgt gaatatcctc ctcttcatgg 1200

gttattcaac aaagccacaa ctggctattg ttacccagtg gtgtgagggc tccagtttat 1260

atcatcatct ccacatcatt gagaccaaat tcgagatgat caaacttata gatattgcac 1320

ggcagactgc acagggcatg gattacttac acgccaagtc aatcatccac agagacctca 1380

agagtaataa tatttttctt catgaagacc tcacagtaaa aataggtgat tttggtctag 1440

ccacagtgaa atctcgatgg agtgggtccc atcagtttga acagttgtct ggatccattt 1500

tgtggatggc accagaagta atcagaatgc aagataaaaa cccatatagc tttcagtcag 1560

atgtatatgc atttgggatt gttctgtatg aattgatgac cggacagtta ccttattcaa 1620

atatcaacaa cagggaccag ataattttta tggtgggacg aggatatctg tctccagatc 1680

tcagtaaggt acggagtaac tgtccaaaag ccatgaagag attaatggca gagtgcctaa 1740

aaaagaaaag agatgaaaga ccactctttc cccaagtagg aaagactctc ctaagcaaga 1800

gacaaaattc agaagttatc agggaaaaag ataagcagat tctcgcctct attgagctgc 1860

tggcccgctc attgccaaaa attcaccgca gtgcatcaga accctccttg aatcgggctg 1920

gcttccaaac agaggatttt agtctatatg cttgtgcttc tccaaaaaca cccattcagg 1980

cagggggata tggagaattt gcagccttca agtagccaca ccatcatgac agcatctact 2040

cttatttctt aagtcttgtg ttcgtacaat ttgttaacat caaaacacag ttctgttcct 2100

caactctttt taaagttaaa atttttcagt gcataagctg gtgtggaaca gaaggaaatt 2160

tcccatccaa caaaagaggg aagaatgttt taggaaccag aattctctgc tgccagtgtt 2220

tcttcttcaa cacaaatatc acaagtctgc ccactcccag gaagaaagag gagagaccct 2280

gagttctgac cttttgatgg tcaggcatga tggaaagaaa ctgctgctac agcttgggag 2340

atttgctctg ggaagtctgc cagtcaactt tgcccttcta accaccagat caatatgtgg 2400

ctgatcatct gatggggcag ttgcaatcac caagccttgt tctctttcct gttctgggat 2460

tgtgttgtgg aacccttttc cctagccacc accagttcat ttctgaggga tggaacaaaa 2520

atgcagcatg cccttcctgt gtggtgcatg ttcagtcctt gacaaatttt taccaaaatg 2580

aagctacttt atttaaaagg agggtgagag gtgaggaggt cactttgggt gtggcggaaa 2640

gggaatgctg catctttttc ctgggctgct ggggctctgg ccttggcttg ccagccggaa 2700

gcgctggcac gcatcgcctt cttttcccat tgggtccagc aatgaagacg agtgtttggg 2760

gttttttttt tctccaccat gtagcaagtt ctcaggaaaa tacaattgat atcttcctcc 2820

taagctcttc caatcagtca ccaagtactt atgtggttac tttgtccagg gcacaaaatg 2880

cctgtatcta attaaaagcc tacaaaactg cttgataaca gttttgaatg tgagacattt 2940

atgtaattta aatgtaaggt acaagtttta atttctgagt ttcttctatt atatttttat 3000

taaaaaaaga aaataatttt cagattgaat tggagtaaaa taatattact tcccactaga 3060

attatatatc ctggaaaatt gtatttttgt tacataagca gcttttaaag aaagatcatt 3120

acccttttct ctacataaat atatggggag tcttagccta atgacaaata tttataattt 3180

ttaaattaat ggtacttgct ggatccatac taacatcttt actaatacct cattgtttct 3240

tccaacttac tcctacacta catcctacat cttcttccta gtcttttatc tagaatatgc 3300

aacctcaaat aaaaatggtg gtgtcctcat tcattctcct ccttcctttt ttcccaagcc 3360

tgatcttcaa aaggttggtt aatttggcag ctgagttcct ccccaggcag agaatagacc 3420

aattttaggt gtattgggac tgagggagga tgtgtaaaga ttaacatcag taaagaaccg 3480

ctgtggagta attaagaact ttgttcttta taactggaga atataaccta accctaacat 3540

ccctcagcct ttactaaagt gtggcgtaaa tcacagtagt agcaaagaaa gtgactctgg 3600

atgtgttcct ggccagtacc tcccttatca tgaatgtaga ctctctcatc aagatttagg 3660

aatataaatc aaatcaaatg tgcccagcca agctatgtag taagggactt gaacaatatt 3720

aggcagaacc tataaaataa atcagggaat tagaaattat ttaaagtttt caaattgtaa 3780

attgccccgg tgtctttcag cctactgcca ttatttttgc tacaatacct acatttcaga 3840

ggagggccta ctgaaaattc catgcaagtg gaaaataatc ctcaagttat taatgagttt 3900

gaaaagcaat gagttcttaa gtctttgtga gtagagcaag atcctacaaa attcagaaat 3960

agtaaaaatg gattcatgct gatttgaaga gcatctgtgt gcataatata atgctgcatc 4020

tcttttaaaa gcagtctatt tttcttttta aatttgtccc catagatgct tttgaacatg 4080

aacatgctta tgttaccttt tccgaggttg ggaagagcca ggagctctca ggcagggccc 4140

cctccctcag ctgggcagga gctgctcagg aggagctagt tatagaggaa gcttagcgtt 4200

ggcattttca aaattcaagg tgataacgct ttcttcttcc tttctgtttt agaatagatt 4260

gctgtctgat ttgaaaaagg gaaatagatt tgatctcaaa tgaatctgtg cccagaagcc 4320

aggctcaggg tattcagaga tttgtatagt gccctcaaaa aataacaaaa ttttagcttt 4380

ccttttttct tcttttctcc atcaaattct tttttctcta gtttacaaat gacatggaaa 4440

aggaatttcc cctgagtttt gtatgccttt ttttttttgg cttagactat agataggcgt 4500

gttgagctcc taagaaaata caaggaggaa ctctttgttg tgcagagcac tttatgagta 4560

gtttgtgtgg ataatatgtg actgcttccc tgacgagctt gtgaggctgt acttatgtct 4620

ttcctgtaag gcagcttcag tgccttctgt agtgtatata aggaaagatt acgccttctg 4680

aaaaatctca gagcaaccat aagattattt taaaatatgt agtatgactg atggactttt 4740

tcatcattaa attagtctag catctaaact tttaccactg aaataatatt gaccaaaaag 4800

caatttataa aaggtatttg tgaatagaaa atacaatgtg atcatttgta cttatgtgca 4860

ccttaaaaga ggaattctgt ctagctgtca aattctggtt ccttaacatc cagtccttga 4920

ttgtgattga gatctggtag gacgtgctgg ggcacgctag cagataaaat cccgtatact 4980

ttaggataga tgttacattt atgtcagtgt tggcaaagag cattgtgtag taataaagaa 5040

ttcaagactt cagcaatgtc aacctgaaac tttgtaaata tttcctagat tgttatttga 5100

tgcagtcaca gctctttatc acacaatgtt gtctttccct catcaggcaa ttttagaact 5160

gctgcacacc cctcctcaga tctcacctgc ccctcctgta cattcacctc tccagccttg 5220

tgcacacctc atttagcttt agtttgaaac acattgcagg gttcaggtga cctcttcaaa 5280

aactacctcc tcagaatgag gtaatgaata gttatttatt ttaaaatatg aaaagtcagg 5340

agctctagaa tatgaagatg atctaagatt ttaactttta tgtatacttg ttgagcactc 5400

tccttttgtc ctaaagggca ttatacattt aagcagtaat actgaaaaat gtagctcaga 5460

gtaactgaat gttgttgaaa gtggtgccag aatctgtttt aggggtacgt atcagaatct 5520

taatcttaaa tcggttacat gaaattaaat agttaatggt aacacttgac taacagatat 5580

aattttaatt ttcggtaggc ttttagcaag acagtaagta catcttcata atgagttagc 5640

cacagcttca tcacatgcac agattttcct gttgagagac tgcccagtta agagggtaga 5700

atgatgaacc atttttcagg attctcttct ttgtccaaac tggcattgtg agtgctagaa 5760

tatcagcact ttcaaactag tgattccaac tattaggcta ttaaaaagca aaacaaacca 5820

aacaaaccat agccagacat gggaagttta ctatgagtat aaacagcaaa tagcttacag 5880

gtcatacatt gaaatggtgt aggtaaggcg ttagaaaaat accttgacaa tttgccaaat 5940

gatcttactg tgccttcatg atgcaataaa aaaaaaaaaa atttagcata aatcagtgat 6000

ttgtgaagag agcagccacc ctggtctaac tcagctgtgt taatattttt tagcgtgcaa 6060

tttagactgc aaagataaat gcactaaaga gtttatagcc aaaatcacat ttaaaaaatg 6120

agagaaaaca caggtaaatt ttcagtgaac aaaattattt ttttaaagta cataatccct 6180

agtatagtca gatatattta tcacatagag caaataggtt gaaatcacaa ttcagtgaca 6240

tttctagaga aactttttct actcccatag gttcttcaaa gcatggaact tttatataac 6300

agaaatgtgt gacggtcatt ttaaattgct gtagtttggg gctgaagtac tgtgtgctgg 6360

gcagcaatca catgtattaa ctagtgagaa aggagaaatt aagatatagg acagaatttg 6420

attttcttgt tcccagatta ctgctgccaa cctagacact gagtttccag aggctgaaac 6480

gtaaacttgc agctcagcaa ctgttttgca aagttagtgg gactgtcctg cttatgctgt 6540

tcaaaaatgc tctgagggcc aggtggggcc tccaggggct cctctctgag gggacatcag 6600

actagctaac gacctggcgg gcggatgtga accggacaca ctccatggtg tgcttcttgt 6660

atcggtccct cgccaccctc aagaaaggct tcagcgggtt ctctagacgt ctccactaag 6720

gtgtgttact aacagccatg ggttgttgag cacccgagga gtgcaatagc atctctgcat 6780

gattgtatat tggcccgaag agaatgaagt ggccagtgta ctcatgttcc atgttgctag 6840

ctctggtaaa ctgaaaatac tggtaagatt tttgttttat cagtacacta gagagtaagc 6900

tttgttttgt tgtttttaga taatgttttc acttccattt ggaaagacat ttaaattgag 6960

tttcagtcct aaattttgcc agtcatggta attagcagtt tctatcaggt atttttaagg 7020

tagaagagga tagaaacata agttctaaaa gcttaaggta accgtggttt attttaaaat 7080

gtttaggggt ggttagtctc tacctcaaaa aaagtgagtg aatcttttat ttcagcattc 7140

acaagttcgg ctgttgtttt tgtaatacat ttttttttta accttttgac ccccctttac 7200

ctaagtgtca atgtagtttt attaattact aagtcagttt cattaaaatg tttatttagc 7260

agttttgact aattgcaatg attaatatag ccagttgtgc atgaggacac agccagtgag 7320

tatatctggg ttttttttgt gatgcttttt ttcttaagac ttctgtagat ttatgaagta 7380

ctcattgaaa acaactaaaa tacgtttatt cgtgttaata tggaaaaaaa aaaa 7434

<210>40

<211>603

<212>PRT

<213> cattle

<400>40

Met Met Arg Gly Leu Ile Pro Glu Cys Cys Ala Val Tyr Arg Ile Gln

1 5 10 15

Asp Gly Glu Lys Lys Pro Ile Gly Trp Asp Thr Asp Ile Ser Trp Leu

20 25 30

Thr Gly Glu Glu Leu His Val Glu Val Leu Glu Asn Val Pro Leu Thr

35 40 45

Thr His Asn Phe Val Arg Lys Thr Phe Phe Thr Leu Ala Phe Cys Asp

50 55 60

Phe Cys Arg Lys Leu Leu Phe Gln Gly Phe Arg Cys Gln Thr Cys Gly

65 70 75 80

Tyr Lys Phe His Gln Arg Cys Ser Thr Glu Val Pro Leu Met Cys Val

85 90 95

Asn Tyr Asp Gln Leu Asp Leu Leu Phe Val Ser Lys Phe Phe Glu His

100 105 110

His Pro Ile Pro Gln Glu Glu Ala Ser Leu Ala Glu Thr Thr Leu Pro

115 120 125

Cys Gly Ser Ser Pro Ser Ala Pro Pro Ser Asp Ser Ile Gly Pro Pro

130 135 140

Ile Leu Thr Ser Pro Ser Pro Ser Lys Ser Ile Pro Ile Pro Gln Pro

145 150 155 160

Phe Arg Pro Ala Asp Glu Asp His Arg Asn Gln Phe Gly Gln Arg Asp

165 170 175

Arg Ser Ser Ser Ala Pro Asn Val His Ile Asn Thr Ile Glu Pro Val

180 185 190

Asn Ile Asp Asp Leu Ile Arg Asp Gln Gly Phe Arg Ser Asp Gly Gly

195 200 205

Ser Thr Thr Gly Leu Ser Ala Thr Pro Pro Ala Ser Leu Pro Gly Ser

210 215 220

Leu Ser Asn Val Lys Ala Leu Gln Lys Ser Pro Gly Pro Gln Arg Glu

225 230 235 240

Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn Arg Met Lys Thr Leu

245 250 255

Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly Gln Ile

260 265 270

Thr Val Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr Val Tyr Lys

275 280 285

Gly Lys Trp His Gly Asp Val Ala Val Lys Met Leu Asn Val Thr Ala

290 295 300

Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val Gly Val Leu

305 310 315 320

Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met Gly Tyr Ser Thr

325 330 335

Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu Gly Ser Ser Leu

340 345 350

Tyr His His Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys Leu

355 360 365

Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His Ala

370 375 380

Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe Leu His

385 390 395 400

Glu Asp Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr Val Lys

405 410 415

Ser Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser Gly Ser Ile

420425 430

Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys Asn Pro Tyr

435 440 445

Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu Tyr Glu Leu

450 455 460

Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln Ile

465 470 475 480

Ile Phe Met Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys Val

485 490 495

Arg Ser Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys Leu

500 505 510

Lys Lys Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Val Gly Lys Thr

515 520 525

Leu Leu Ser Lys Arg Gln Asn Ser Glu Val Ile Arg Glu Lys Asp Lys

530 535 540

Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro Lys Ile

545 550 555 560

His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe Gln Thr

565 570 575

Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys Thr Pro Ile Gln

580 585 590

Ala Gly Gly Tyr Gly Glu Phe Ala Ala Phe Lys

595 600

<210>41

<211>2295

<212>DNA

<213> horse (Equus caballus)

<220>

<221>misc_feature

<222>(79)..(79)

<223> n is a, c, g, or t

<400>41

atgaagacgc tgagcggcgg cggcggcggc gcggagcagg gccaggctct gttcaacggg 60

gacatggaac ccggaggcnc cgcgccggcg cccgcggcct cgtcggccgc ggaccctgcc 120

attcccgagg aggtatggaa tatcaaacaa atgattaaat tgacacagga acatatagag 180

gccctattgg acaaatttgg tggggagcat aatccaccat caatatatct ggaggcctat 240

gaagaataca ccagcaagct agatgccctc caacaaagag aacaacagtt attggaatcc 300

ctggggaatg gaactgattt ttctgtttct agttctgcat caacggacac cgttacatct 360

tcttcctctt ctagcctttc agtgctacct tcatctcttt cagtttttca aaatcccaca 420

gatgtgtcac ggagcaaccc taagtcacca caaaaaccta tcgttagagt cttcctgccc 480

aacaaacaga ggacagtggt acctgcaagg tgtggcgtta cagtccggga cagtctaaag 540

aaagcactga tgatgagagg tctaatccca gagtgctgtg ctgtttacag aattcaggat 600

ggagagaaga aaccaattgg ctgggacact gatatttcct ggctcactgg agaggaattg 660

catgtagaag tgttggagaa tgttccactt acaacacaca actttgtacg gaaaactttt 720

ttcaccttag cattttgtga cttttgtcga aagctgcttt tccagggttt ccgctgtcaa 780

acatgtggtt ataaatttca ccagcgttgt agtacagagg ttccactgat gtgtgttaat 840

tatgaccaac ttgatttgct gtttgtctcc aagttctttg aacaccaccc agtatcacag 900

gaggaggcct ccttagcaga gactgccctt acatctggat catccccttc tgcacccccc 960

tccgattcca ttgggcccca aattctcacc agtccatctc cttcaaaatc cattccaatt 1020

ccacagcctt tccgaccagc agatgaagat catcgaaatc agtttggaca acgagaccgg 1080

tcctcatcag ctccaaatgt acatataaac acaatagaac ctgtcaatat tgatgacttg 1140

attagagacc aagggtttcg tagtgatgga ggatcaacca caggtttatc tgccaccccc 1200

cctgcctcat tacctggctc actcactaat gtgaaggcat tacagaaatc tccaggacct 1260

caacgggaaa ggaaatcatc ttcatcctca gaagacagga atcgaatgaa aactcttggt 1320

agacgggatt caagtgacga ttgggagatt cctgatgggc agatcacagt gggacaaaga 1380

attggatctg ggtcatttgg gacagtctac aagggaaagt ggcatggtga tgtggcagtg 1440

aaaatgttga atgtgacagc acccacacct cagcagttac aggccttcaa aaatgaagta 1500

ggagtactca ggaaaactcg acatgtgaat atcctactct tcatgggcta ttcaacaaag 1560

ccacaactgg ctattgttac ccagtggtgt gagggctcca gcttatatca ccatctccac 1620

atcattgaga ccaaatttga gatgatcaaa cttatagata ttgctcggca aactgcacag 1680

ggcatggatt acttacacgc caagtcaatc atccacagag acctcaagag taataatatt 1740

tttcttcatg aagacctcac agtaaaaata ggtgattttg gtctagccac agtgaaatct 1800

cgatggagtg ggtcccatca gtttgaacag ttgtctggat ccattttgtg gatggcacca 1860

gaagtaatca gaatgcaaga taaaaacccg tatagctttc aatcagatgt atatgccttt 1920

gggattgttc tgtatgaatt gatgactgga cagttacctt attcaaacat caacaacagg 1980

gaccagataa tttttatggt gggaagagga tatctatctc cagatctcag taaggtacgg 2040

agtaactgtc caaaagccat gaagagatta atggcagagt gcctaaaaaa gaaaagagac 2100

gagagaccac tcttccccca aattctcgcc tctattgagc tgctggcccg ctcattgcca 2160

aaaattcacc gcagtgcatc agagccctcc ttgaatcggg ctggcttcca gacagaggat 2220

tttagtctat atgcttgtgc ttctccgaaa acacccatcc aggcaggggg atatggtgcg 2280

tttcctgtcc actga 2295

<210>42

<211>764

<212>PRT

<213> horse

<220>

<221>misc_feature

<222>(27)..(27)

<223> Xaa can be any naturally occurring amino acid

<400>42

Met Lys Thr Leu Ser Gly Gly Gly Gly Gly Ala Glu Gln Gly Gln Ala

1 5 10 15

Leu Phe Asn Gly Asp Met Glu Pro Gly Gly Xaa Ala Pro Ala Pro Ala

20 25 30

Ala Ser Ser Ala Ala Asp Pro Ala Ile Pro Glu Glu Val Trp Asn Ile

35 40 45

Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu Leu Asp

50 55 60

Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu Ala Tyr

65 70 75 80

Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu Gln Gln

85 90 95

Leu Leu Glu Ser Leu Gly Asn Gly Thr Asp Phe Ser Val Ser Ser Ser

100 105 110

Ala Ser Thr Asp Thr Val Thr Ser Ser Ser Ser Ser Ser Leu Ser Val

115 120 125

Leu Pro Ser Ser Leu Ser Val Phe Gln Asn Pro Thr Asp Val Ser Arg

130 135 140

Ser Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe Leu Pro

145 150 155 160

Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr Val Arg

165 170 175

Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro Glu Cys

180 185 190

Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile Gly Trp

195 200 205

Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val Glu Val

210 215 220

Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys Thr Phe

225 230 235 240

Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe Gln Gly

245 250 255

Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys Ser Thr

260 265 270

Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu Leu Phe

275 280 285

Val Ser Lys Phe Phe Glu His His Pro Val Ser Gln Glu Glu Ala Ser

290 295 300

Leu Ala Glu Thr Ala Leu Thr Ser Gly Ser Ser Pro Ser Ala Pro Pro

305 310 315 320

Ser Asp Ser Ile Gly Pro Gln Ile Leu Thr Ser Pro Ser Pro Ser Lys

325 330 335

Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp His Arg

340 345 350

Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn Val His

355 360 365

Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg Asp Gln

370 375 380

Gly Phe Arg Ser Asp Gly Gly Ser Thr Thr Gly Leu Ser Ala Thr Pro

385 390 395 400

Pro Ala Ser Leu Pro Gly Ser Leu Thr Asn Val Lys Ala Leu Gln Lys

405 410 415

Ser Pro Gly Pro Gln Arg Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp

420 425 430

Arg Asn Arg Met Lys Thr Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp

435 440 445

Glu Ile Pro Asp Gly Gln Ile Thr Val Gly Gln Arg Ile Gly Ser Gly

450 455 460

Ser Phe Gly Thr Val Tyr Lys Gly Lys Trp His Gly Asp Val Ala Val

465 470 475 480

Lys Met Leu Asn Val Thr Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe

485 490 495

Lys Asn Glu Val Gly Val Leu Arg Lys Thr Arg His Val Asn Ile Leu

500 505510

Leu Phe Met Gly Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr Gln

515 520 525

Trp Cys Glu Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu Thr

530 535 540

Lys Phe Glu Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln

545 550 555 560

Gly Met Asp Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu Lys

565 570 575

Ser Asn Asn Ile Phe Leu His Glu Asp Leu Thr Val Lys Ile Gly Asp

580 585 590

Phe Gly Leu Ala Thr Val Lys Ser Arg Trp Ser Gly Ser His Gln Phe

595 600 605

Glu Gln Leu Ser Gly Ser Ile Leu Trp Met Ala Pro Glu Val Ile Arg

610 615 620

Met Gln Asp Lys Asn Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe

625 630 635 640

Gly Ile Val Leu Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn

645 650 655

Ile Asn Asn Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr Leu

660 665 670

Ser Pro Asp Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met Lys

675 680 685

Arg Leu Met Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg Pro Leu

690 695 700

Phe Pro Gln Ile Leu Ala Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro

705 710 715 720

Lys Ile His Arg Ser Ala Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe

725 730 735

Gln Thr Glu Asp Phe Ser Leu Tyr Ala Cys Ala Ser Pro Lys Thr Pro

740 745 750

Ile Gln Ala Gly Gly Tyr Gly Ala Phe Pro Val His

755 760

<210>43

<211>2678

<212>DNA

<213> Chicken (Gallus Gallus)

<400>43

tccccctccc tcgccccagc gcttcgatcc aagatggcgg cgctgagcag cggcagcagc 60

gccgaggggg cctcgctctt caacggggac atggagcccg agccgccgcc gcccgtgctg 120

ggcgcctgct acgccgggag cggcggcggc gacccggcca tcccggagga ggtgtggaat 180

atcaaacaga tgattaaatt aacacaagaa catatagaag cgctgttaga caagtttgga 240

ggagagcata acccaccatc aatatattta gaggcctatg aggagtacac cagcaaacta 300

gatgctctac agcagagaga acagcagtta ttggaatcca tgggaaatgg aactgatttc 360

tctgtttcca gttcagcttc aacggacaca gttgcatcat cttcctcctc tagcctctct 420

gtagcacctt catccctttc agtttatcaa aatcctactg atatgtcgcg gaataaccct 480

aagtctccac agaagcctat tgttagagtc ttcctgccca acaagcaaag gactgtggtt 540

ccggcaagat gtggggtgac agtccgagac agcctgaaga aagctctgat gatgagaggt 600

cttattccag aatgctgtgc tgtttacaga atacaggatg gagagaagaa gccaattggc 660

tgggacactg acatttcctg gctaaccgga gaggagttac acgtggaggt cttggagaat 720

gtgccactca caacacacaa ttttgtacga aaaacattct tcacgttagc gttctgcgac 780

ttctgtcgaa agctgctttt ccagggattc cgatgccaga catgtggcta caaatttcac 840

cagcgctgta gcacagaagt gccactgatg tgtgttaact acgaccaact cgatttgctg 900

tttgtctcca agttctttga acatcacccc atatcgcagg aggagaccac cttaggagag 960

accaccccgg catcgggatc gtacccctca gtgcccccat cagattctgt tggaccacca 1020

attctcccta gtccttctcc ttcaaaatcc attccaatcc cacagccctt ccgaccagca 1080

gatgaagacc atcggaatca gtttgggcaa cgcgaccgat cctcttcagc tcccaatgtt 1140

cacatcaata caattgagcc agtcaatatt gatgacttga ttagagacca gggtgtacga 1200

ggagagggag cccctttgaa ccagctgatg cgctgtcttc ggaaatacca atcccggact 1260

cccagtcccc tccttcattc tgtccccagt gaaatagtgt ttgattttga gcctggccca 1320

gtgttcagag gttcaactgc aggtttgtct gcaacacctc ctgcatcttt gcctgggtca 1380

cttaccaatg tgaaagcatt acagaaatca ccaggccccc aacgggaaag gaaatcatcc 1440

tcatcctcag aagacagaaa taggatgaaa acccttggtc gacgagattc aagtgatgat 1500

tgggaaatac cagatgggca gatcacagtt ggacaaagga taggatctgg atcatttgga 1560

acagtctaca aaggaaagtg gcatggtgac gtggcagtga aaatgttgaa tgttacagca 1620

cccacacctc aacagttaca ggctttcaaa aatgaagtag gagtgctcag gaaaacacgg 1680

catgtgaata tcctactttt tatgggttat tcaacaaaac ctcagttggc tattgttaca 1740

cagtggtgtg aggggtccag cttatatcac catctgcaca taattgagac caagtttgaa 1800

atgatcaaac taattgatat tgcacgacag actgcacaag gcatggatta tttgcatgcc 1860

aagtcaatca tccacagaga cctcaagagt aataatattt ttcttcatga agacctcaca 1920

gtaaaaatag gtgacttcgg tctggctaca gtgaaatcac gatggagtgg atctcatcaa 1980

tttgaacagt tatctggatc aattctatgg atggcaccgg aagtgatcag gatgcaagac 2040

aaaaacccat atagctttca gtcagatgtg tatgcattcg ggattgtgct ttatgaactg 2100

atgactggac agttaccata ctcaaacatc aacaacaggg accagataat ttttatggtg 2160

ggacgaggat atctatctcc agacctcagt aaagtaagaa gtaactgtcc aaaagctatg 2220

aagagactaa tggcagaatg cttgaaaaag aaaagagatg agagacctct ttttccacag 2280

attcttgcct ccattgagct tctggcccgg tcgttgccaa aaattcaccg cagtgcatct 2340

gagccgtcac taaaccgggc tggcttccag accgaggatt tcagtctgta tgcttgtgct 2400

tctccaaaaa cgcccatcca agcaggggga tacggtgggt ttccagtaca ctgaaaagaa 2460

atgtgaaagc gtgtgcctgt ttgctcatgt gctggtgtgt tcctgtgtgt gcaacgcata 2520

cgtacgttct cagttcctac cagcgacttt ttaaggttta ctgagggaat gaagactcat 2580

ttcctaacat ggggcattga acgtcctgag cacaagtcag tgctggtaag gaatgtcttg 2640

ggaacagctg gcaagaagaa ttagaaggta cttaaagg 2678

<210>44

<211>806

<212>PRT

<213> Chicken

<400>44

Met Ala Ala Leu Ser Ser Gly Ser Ser Ala Glu Gly Ala Ser Leu Phe

1 5 10 15

Asn Gly Asp Met Glu Pro Glu Pro Pro Pro Pro Val Leu Gly Ala Cys

20 25 30

Tyr Ala Gly Ser Gly Gly Gly Asp Pro Ala Ile Pro Glu Glu Val Trp

35 40 45

Asn Ile Lys Gln Met Ile Lys Leu Thr Gln Glu His Ile Glu Ala Leu

50 55 60

Leu Asp Lys Phe Gly Gly Glu His Asn Pro Pro Ser Ile Tyr Leu Glu

65 70 75 80

Ala Tyr Glu Glu Tyr Thr Ser Lys Leu Asp Ala Leu Gln Gln Arg Glu

85 90 95

Gln Gln Leu Leu Glu Ser Met Gly Asn Gly Thr Asp Phe Ser Val Ser

100 105 110

Ser Ser Ala Ser Thr Asp Thr Val Ala Ser Ser Ser Ser Ser Ser Leu

115 120 125

Ser Val Ala Pro Ser Ser Leu Ser Val Tyr Gln Asn Pro Thr Asp Met

130 135 140

Ser Arg Asn Asn Pro Lys Ser Pro Gln Lys Pro Ile Val Arg Val Phe

145 150 155 160

Leu Pro Asn Lys Gln Arg Thr Val Val Pro Ala Arg Cys Gly Val Thr

165 170 175

Val Arg Asp Ser Leu Lys Lys Ala Leu Met Met Arg Gly Leu Ile Pro

180 185 190

Glu Cys Cys Ala Val Tyr Arg Ile Gln Asp Gly Glu Lys Lys Pro Ile

195 200 205

Gly Trp Asp Thr Asp Ile Ser Trp Leu Thr Gly Glu Glu Leu His Val

210 215 220

Glu Val Leu Glu Asn Val Pro Leu Thr Thr His Asn Phe Val Arg Lys

225 230 235 240

Thr Phe Phe Thr Leu Ala Phe Cys Asp Phe Cys Arg Lys Leu Leu Phe

245 250 255

Gln Gly Phe Arg Cys Gln Thr Cys Gly Tyr Lys Phe His Gln Arg Cys

260 265 270

Ser Thr Glu Val Pro Leu Met Cys Val Asn Tyr Asp Gln Leu Asp Leu

275 280 285

Leu Phe Val Ser Lys Phe Phe Glu His His Pro Ile Ser Gln Glu Glu

290 295 300

Thr Thr Leu Gly Glu Thr Thr Pro Ala Ser Gly Ser Tyr Pro Ser Val

305 310 315 320

Pro Pro Ser Asp Ser Val Gly Pro Pro Ile Leu Pro Ser Pro Ser Pro

325 330 335

Ser Lys Ser Ile Pro Ile Pro Gln Pro Phe Arg Pro Ala Asp Glu Asp

340 345 350

His Arg Asn Gln Phe Gly Gln Arg Asp Arg Ser Ser Ser Ala Pro Asn

355 360 365

Val His Ile Asn Thr Ile Glu Pro Val Asn Ile Asp Asp Leu Ile Arg

370 375 380

Asp Gln Gly Val Arg Gly Glu Gly Ala Pro Leu Asn Gln Leu Met Arg

385 390 395 400

Cys Leu Arg Lys Tyr Gln Ser Arg Thr Pro Ser Pro Leu Leu His Ser

405 410 415

Val Pro Ser Glu Ile Val Phe Asp Phe Glu Pro Gly Pro Val Phe Arg

420 425 430

Gly Ser Thr Ala Gly Leu Ser Ala Thr Pro Pro Ala Ser Leu Pro Gly

435 440 445

Ser Leu Thr Asn Val Lys Ala Leu Gln Lys Ser Pro Gly Pro Gln Arg

450 455 460

Glu Arg Lys Ser Ser Ser Ser Ser Glu Asp Arg Asn Arg Met Lys Thr

465 470 475 480

Leu Gly Arg Arg Asp Ser Ser Asp Asp Trp Glu Ile Pro Asp Gly Gln

485 490 495

Ile Thr Val Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr Val Tyr

500 505 510

Lys Gly Lys Trp His Gly Asp Val Ala Val Lys Met Leu Asn Val Thr

515 520 525

Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val Gly Val

530 535 540

Leu Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met Gly Tyr Ser

545 550 555 560

Thr Lys ProGln Leu Ala Ile Val Thr Gln Trp Cys Glu Gly Ser Ser

565 570 575

Leu Tyr His His Leu His Ile Ile Glu Thr Lys Phe Glu Met Ile Lys

580 585 590

Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly Met Asp Tyr Leu His

595 600 605

Ala Lys Ser Ile Ile His Arg Asp Leu Lys Ser Asn Asn Ile Phe Leu

610 615 620

His Glu Asp Leu Thr Val Lys Ile Gly Asp Phe Gly Leu Ala Thr Val

625 630 635 640

Lys Ser Arg Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser Gly Ser

645 650 655

Ile Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys Asn Pro

660 665 670

Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu Tyr Glu

675 680 685

Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn Arg Asp Gln

690 695 700

Ile Ile Phe Met Val Gly Arg Gly Tyr Leu Ser Pro Asp Leu Ser Lys

705 710 715 720

Val Arg Ser Asn Cys Pro Lys Ala Met Lys Arg Leu Met Ala Glu Cys

725 730 735

Leu Lys Lys Lys Arg Asp Glu Arg Pro Leu Phe Pro Gln Ile Leu Ala

740 745 750

Ser Ile Glu Leu Leu Ala Arg Ser Leu Pro Lys Ile His Arg Ser Ala

755 760 765

Ser Glu Pro Ser Leu Asn Arg Ala Gly Phe Gln Thr Glu Asp Phe Ser

770 775 780

Leu Tyr Ala Cys Ala Ser Pro Lys Thr Pro Ile Gln Ala Gly Gly Tyr

785 790 795 800

Gly Gly Phe Pro Val His

805

<210>45

<211>4454

<212>DNA

<213> human

<400>45

gaaacgtccc gtgtgggagg ggcgggtctg ggtgcggcct gccgcatgac tcgtggttcg 60

gaggcccacg tggccggggc ggggactcag gcgcctgggg cgccgactga ttacgtagcg 120

ggcggggccg gaagtgccgc tccttggtgg gggctgttca tggcggttcc ggggtctcca 180

acatttttcc cggctgtggt cctaaatctg tccaaagcag aggcagtgga gcttgaggtt 240

cttgctggtg tgaaatgact gagtacaaac tggtggtggt tggagcaggt ggtgttggga 300

aaagcgcact gacaatccag ctaatccaga accactttgt agatgaatat gatcccacca 360

tagaggattc ttacagaaaa caagtggtta tagatggtga aacctgtttg ttggacatac 420

tggatacagc tggacaagaa gagtacagtg ccatgagaga ccaatacatg aggacaggcg 480

aaggcttcct ctgtgtattt gccatcaata atagcaagtc atttgcggat attaacctct 540

acagggagca gattaagcga gtaaaagact cggatgatgt acctatggtg ctagtgggaa 600

acaagtgtga tttgccaaca aggacagttg atacaaaaca agcccacgaa ctggccaaga 660

gttacgggat tccattcatt gaaacctcag ccaagaccag acagggtgtt gaagatgctt 720

tttacacact ggtaagagaa atacgccagt accgaatgaa aaaactcaac agcagtgatg 780

atgggactca gggttgtatg ggattgccat gtgtggtgat gtaacaagat acttttaaag 840

ttttgtcaga aaagagccac tttcaagctg cactgacacc ctggtcctga cttccctgga 900

ggagaagtat tcctgttgct gtcttcagtc tcacagagaa gctcctgcta cttccccagc 960

tctcagtagt ttagtacaat aatctctatt tgagaagttc tcagaataac tacctcctca 1020

cttggctgtc tgaccagaga atgcacctct tgttactccc tgttattttt ctgccctggg 1080

ttcttccaca gcacaaacac acctctgcca ccccaggttt ttcatctgaa aagcagttca 1140

tgtctgaaac agagaaccaa accgcaaacg tgaaattcta ttgaaaacag tgtcttgagc 1200

tctaaagtag caactgctgg tgattttttt tttcttttta ctgttgaact tagaactatg 1260

ctaatttttg gagaaatgtc ataaattact gttttgccaa gaatatagtt attattgctg 1320

tttggtttgt ttataatgtt atcggctcta ttctctaaac tggcatctgc tctagattca 1380

taaatacaaa aatgaatact gaattttgag tctatcctag tcttcacaac tttgacgtaa 1440

ttaaatccaa ctttcacagt gaagtgcctt tttcctagaa gtggtttgta gacttccttt 1500

ataatatttc agtggaatag atgtctcaaa aatccttatg catgaaatga atgtctgaga 1560

tacgtctgtg acttatctac cattgaagga aagctatatc tatttgagag cagatgccat 1620

tttgtacatg tatgaaattg gttttccaga ggcctgtttt ggggctttcc caggagaaag 1680

atgaaactga aagcacatga ataatttcac ttaataattt ttacctaatc tccacttttt 1740

tcataggtta ctacctatac aatgtatgta atttgtttcc cctagcttac tgataaacct 1800

aatattcaat gaacttccat ttgtattcaa atttgtgtca taccagaaag ctctacattt 1860

gcagatgttc aaatattgta aaactttggt gcattgttat ttaatagctg tgatcagtga 1920

ttttcaaacc tcaaatatag tatattaaca aattacattt tcactgtata tcatggtatc 1980

ttaatgatgt atataattgc cttcaatccc cttctcaccc caccctctac agcttccccc 2040

acagcaatag gggcttgatt atttcagttg agtaaagcat ggtgctaatg gaccagggtc 2100

acagtttcaa aacttgaaca atccagttag catcacagag aaagaaattc ttctgcattt 2160

gctcattgca ccagtaactc cagctagtaa ttttgctagg tagctgcagt tagccctgca 2220

aggaaagaag aggtcagtta gcacaaaccc tttaccatga ctggaaaact cagtatcacg 2280

tatttaaaca tttttttttc ttttagccat gtagaaactc taaattaagc caatattctc 2340

atttgagaat gaggatgtct cagctgagaa acgttttaaa ttctctttat tcataatgtt 2400

ctttgaaggg tttaaaacaa gatgttgata aatctaagct gatgagtttg ctcaaaacag 2460

gaagttgaaa ttgttgagac aggaatggaa aatataatta attgatacct atgaggattt 2520

ggaggcttgg cattttaatt tgcagataat accctggtaa ttctcatgaa aaatagactt 2580

ggataacttt tgataaaaga ctaattccaa aatggccact ttgttcctgt ctttaatatc 2640

taaatactta ctgaggtcct ccatcttcta tattatgaat tttcatttat taagcaaatg 2700

tcatattacc ttgaaattca gaagagaaga aacatatact gtgtccagag tataatgaac 2760

ctgcagagtt gtgcttctta ctgctaattc tgggagcttt cacagtactg tcatcatttg 2820

taaatggaaa ttctgctttt ctgtttctgc tccttctgga gcagtgctac tctgtaattt 2880

tcctgaggct tatcacctca gtcatttctt ttttaaatgt ctgtgactgg cagtgattct 2940

ttttcttaaa aatctattaa atttgatgtc aaattaggga gaaagatagt tactcatctt 3000

gggctcttgt gccaatagcc cttgtatgta tgtacttaga gttttccaag tatgttctaa 3060

gcacagaagt ttctaaatgg ggccaaaatt cagacttgag tatgttcttt gaatacctta 3120

agaagttaca attagccggg catggtggcc cgtgcctgta gtcccagcta cttgagaggc 3180

tgaggcagga gaatcacttc aacccaggag gtggaggtta cagtgagcag agatcgtgcc 3240

actgcactcc agcctgggtg acaagagaga cttgtctcca aaaaaaaagt tacacctagg 3300

tgtgaatttt ggcacaaagg agtgacaaac ttatagttaa aagctgaata acttcagtgt 3360

ggtataaaac gtggttttta ggctatgttt gtgattgctg aaaagaattc tagtttacct 3420

caaaatcctt ctctttcccc aaattaagtg cctggccagc tgtcataaat tacatattcc 3480

ttttggtttt tttaaaggtt acatgttcaa gagtgaaaat aagatgttct gtctgaaggc 3540

taccatgccg gatctgtaaa tgaacctgtt aaatgctgta tttgctccaa cggcttacta 3600

tagaatgtta cttaatacaa tatcatactt attacaattt ttactatagg agtgtaatag 3660

gtaaaattaa tctctatttt agtgggccca tgtttagtct ttcaccatcc tttaaactgc 3720

tgtgaatttt tttgtcatga cttgaaagca aggatagaga aacactttag agatatgtgg 3780

ggttttttta ccattccaga gcttgtgagc ataatcatat ttgctttata tttatagtca 3840

tgaactccta agttggcagc tacaaccaag aaccaaaaaa tggtgcgttc tgcttcttgt 3900

aattcatctc tgctaataaa ttataagaag caaggaaaat tagggaaaat attttatttg 3960

gatggtttct ataaacaagg gactataatt cttgtacatt atttttcatc tttgctgttt 4020

ctttgagcag tctaatgtgc cacacaatta tctaaggtat ttgttttcta taagaattgt 4080

tttaaaagta ttcttgttac cagagtagtt gtattatatt tcaaaacgta agatgatttt 4140

taaaagcctg agtactgacc taagatggaa ttgtatgaac tctgctctgg agggagggga 4200

ggatgtccgt ggaagttgta agacttttat ttttttgtgc catcaaatat aggtaaaaat 4260

aattgtgcaa ttctgctgtt taaacaggaa ctattggcct ccttggccct aaatggaagg 4320

gccgatattt taagttgatt attttattgt aaattaatcc aacctagttc tttttaattt 4380

ggttgaatgt tttttcttgt taaatgatgt ttaaaaaata aaaactggaa gttcttggct 4440

tagtcataat tctt 4454

<210>46

<211>189

<212>PRT

<213> human

<400>46

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr

20 25 30

Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly

35 40 45

Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr

50 55 60

Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys

65 70 75 80

Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr

85 90 95

Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val

100 105 110

Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys

115 120 125

Gln Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr

130 135 140

Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val

145 150 155 160

Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Asp Asp

165 170 175

Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met

180 185

<210>47

<211>1326

<212>DNA

<213> rat

<400>47

gccgttcatg gcggtttcgg ggtctccaac agcttctcag gttgaaatcc aaaagcctcc 60

cgaggcgggg tctgcggagt ttgagatttt tgcaggtgtg aaatgactga gtacaaactg 120

gtggtggttg gagcaggtgg cgttgggaaa agtgctttga caatccagct aatccagaac 180

cactttgtgg atgaatatga tcccaccata gaggattctt accgaaaaca agtggtgatt 240

gacggtgaga cctgtctact ggacatactg gacacagctg gacaagagga gtacagtgcc 300

atgagagacc aatacatgag gacaggcgaa gggttcctct gtgtgtttgc catcaataat 360

agcaaatcct ttgcagatat taacctctac agggagcaaa ttaagcgcgt gaaagactct 420

gatgatgtac ccatggtgct ggtagggaac aagtgtgact tgccaacaag gacagttgac 480

acaaagcaag cccacgagct ggccaagagt tatggaattc cattcattga aacctcagcc 540

aagacccgac agggtgtgga ggatgccttt tacacgcttg taagggagat acgccagtac 600

cggatgaaga agctcaacag cagtgaggat ggcactcaag gctgtatggg gctgccctgt 660

gtggtgatgt agtaagaccc tttaaaagtt ctgtcatcag aaacgagcca ctttcaagcc 720

tcactgatgc cctggttctg acatccctgg aggagacgtg tttctgctgc tctctgcatc 780

tcagagaagc tcctgcttcc tgcttcccca acttagttac tgagcacagc catctaacct 840

gagacctctt cagaataact acctcctcac tcggctgtcc gaccagagaa atgaacctgt 900

ttctccccag tagttctctg ccctgggttt cccctagaaa caaacacacc tgccagctgg 960

ctttgtcctc cgaaaagcag tttacattga tgcagagaac caaactatag acaagcaatt 1020

ctgttgtcaa cagtttctta agctctaagg taacaattgc tggtgatttc cccctttgcc 1080

cccaactgtt gaacttggcc ttgttagttt tgggggaaat gtcaaaaatt aatctcttcc 1140

cgagaataga attagtgttg ctgattgcct gatttgcaat gtgatcagct atattctata 1200

agctggcgtc tgctctgtat tcataaatgc aaacatgagt actgacgtaa gtgcatccct 1260

agtcttctca gctgcatgca attaaatcca acgttcacaa caaaaaaaaa aaaaaaaaaa 1320

aaaaaa 1326

<210>48

<211>189

<212>PRT

<213> rat

<400>48

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 1015

Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr

20 25 30

Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly

35 40 45

Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr

50 55 60

Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys

65 70 75 80

Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr

85 90 95

Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val

100 105 110

Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys

115 120 125

Gln Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr

130 135 140

Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val

145150 155 160

Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Glu Asp

165 170 175

Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met

180 185

<210>49

<211>4470

<212>DNA

<213> mice

<400>49

gggactgggg cgccttgggc gcctagtgat tacgtagcgg gtggggccgg aagtgccgct 60

ccctggcggg ggctgttcat ggcggtttcg gggtctccaa cagcttctca ggttgaagtc 120

caaaagcctc ccgaggcggg gtctgcggag tttgaggttt ttgctggtgt gaaatgactg 180

agtacaaact ggtggtggtt ggagcaggtg gtgttgggaa aagcgccttg acgatccagc 240

taatccagaa ccactttgtg gatgaatatg atcccaccat agaggattct taccgaaagc 300

aagtggtgat tgatggtgag acctgcctgc tggacatact ggacacagct ggacaagagg 360

agtacagtgc catgagagac cagtacatga ggacaggcga agggttcctc tgtgtatttg 420

ccatcaataa tagcaaatca tttgcagata ttaacctcta cagggagcaa attaagcgtg 480

tgaaagattc tgatgatgtc cccatggtgc tggtaggcaa caagtgtgac ttgccaacaa 540

ggacagttga cacaaagcaa gcccacgaac tggccaagag ttacggaatt ccattcattg 600

agacctcagc caagacccga cagggtgtgg aggatgcctt ttacacactg gtaagggaga 660

tacgccagta ccgaatgaaa aagctcaaca gcagtgacga tggcactcaa ggttgtatgg 720

ggctgccctg tgtgctgatg tagtaagaca ctttgaaagt tctgtcatca gaaaagagcc 780

actttgaagc tgcactgatg ccctggttct gacatccctg gaggagacct gttcctgctg 840

ctctctgcat ctcagagaag ctcctgcttc ctgcttcccc gactcagtta ctgagcacag 900

ccatctaacc tgagacctct tcagaataac tacctcctca ctcggctgtc tgaccagaga 960

aatagacctg tctctcccgg tcgttctctg ccctgggttc ccctagaaac agacacagcc 1020

tccagctggc tttgtcctct gaaaagcagt ttacattgat gcagagaacc aaactagaca 1080

tgccattctg ttgacaacag tttcttatac tctaaggtaa caactgctgg tgattttccc 1140

ctgcccccaa ctgttgaact tggccttgtt ggtttggggg gaaaatgtca taaattactt 1200

tcttcccaaa atataattag tgttgctgat tgatttgtaa tgtgatcagc tatattccat 1260

aaactggcat ctgctctgta ttcataaatg caaacacgaa tactctcaac tgcatgcaat 1320

taaatccaac attcacaaca aagtgccttt ttcctaaaag tgctctgtag gctccattac 1380

agtttgtaat tggaatagat gtgtcaagaa ccattgtata ggaaagtgac tctgagccat 1440

ctacctttga gggaaaggtg tatgtacctg atggcagatg ctttgtgtat gcacatgaag 1500

atagtttccc tgtctgggat tctcccagga gaaagatgga actgaaacaa ttacaagtaa 1560

tttcatttaa ttctagctaa tctttttttt tttttttttt ttttttggta gactatcacc 1620

tataaatatt tggaatatct tctagcttac tgataatcta ataattaatg agcttccatt 1680

ataatgaatt ggttcatacc aggaagccct ccatttatag tatagatact gtaaaaattg 1740

gcatgttgtt actttatagc tgtgattaat gattcctcag accttgctga gatatagtta 1800

ttagcagaca ggttatatct ttgctgcata gtttcttcat ggaatatata tctatctgta 1860

tgtggagaga acgtggccct cagttccctt ctcagcatcc ctcatctctc agcctagaga 1920

agttcgagca tcctagaggg gcttgaacag ttatctcggt taaaccatgg tgctaatgga 1980

ccgggtcatg gtttcaaaac ttgaacaagc cagttagcat cacagagaaa cagtccatcc 2040

atatttgctc cctgcctatt attcctgctt acagactttt gcctgatgcc tgctgttagt 2100

gctacaagga taaagcttgt gtggttctca ccaggactgg aagtacctgg tgagctctgg 2160

ggtaagccta gatatcttta cattttcaga cccttattct tagccacgtg gaaactgaag 2220

ccagagtcca tacctccatc tccttccccc cccaaaaaaa ttagattaat gttctttata 2280

tagctttttt aaagtattta aaacatgtct ataagttagg ctgccaacta acaaaagctg 2340

atgtgtttgt tcaaataaag aggtatcctt cgctactcga gagaagaatg taaaatgcca 2400

ttgattgttg tcacttggag gcttgatgtt tgccctgata attcattagt gggttttgtt 2460

tgtcacatga tacctaagat gtaactcagc tcagtaattc taatgaaaac ataaattgga 2520

taccttaatt gaaaaaagca aacctaattc caaaatggcc attttctctt ctgatcttgt 2580

aatacctaaa attctgaggt ccttgggatt cttttgttta taacaggatc ttgctgtgta 2640

gtcctagctg gcctcaaact cacaatactc ttcctggatc aatctcccaa gtgctgggat 2700

tacaggcaca ttccaccaca cacacctgac tgagctcgtt cctaatgagt tttcattaag 2760

caaattcccc atcaccttga aactaatcag aagggggaag aaacatttgc tatgctcctg 2820

agtgctaaca ctgggatcat tcacatgggg tttgcattcc taggcaaact aaactgctgc 2880

cttttacaac aaggctcagt catcttcctg aagctgctga gaccagcact tggtcttgtt 2940

ttgttttaat atgtctatat gactggtggt ggatccctaa atagtttatt aattaaactc 3000

cagttaagga gaaagttact caccttgacc cgtttgacca tatcccgtgt gtgtgtgtgt 3060

gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgcacgcgt atgtacgtac gtatgtatgt 3120

aggtatgtag gtggtttcca gtataaacac agaaacaaat ggagccaatt caggtttcag 3180

atgcccttac taacatatat tcccacgggg tgtgggtttt ggcacaacag tgacaaactt 3240

aaaagccaag taagagccgg gcgtggtggc gcacgccttt aatcccagca cttgggaggc 3300

agaggcaggc ggatttctga gttctaggcc atcctggtct acagtgagtt ccaggacagc 3360

cagtgctaca cagagaaacc ctgtctcgaa aagccaaaaa aaaaaaaaaa aaaaaaaaaa 3420

aaaagccaag taggtccagt tggtatagta tcaaagtgtt tttagagtaa ttagtgaagg 3480

tctgctttac ctcaaagttg cagagcctct cttcctgagt ttaagtgcct ggccggcagt 3540

cacaaattaa catgttgctg taaggcagtt agttgaagct ttgttcacac attggagagt 3600

atgaaaataa agtgttctaa gagcgctgat actggatctg tgtaaacctg gtaaatgccg 3660

tttgtccagg acttagcgtg tgtgagttgg tagctcagta cgagtttact agttccgcag 3720

tgtgtacaat ggaggcgggt ttgttttagc tggccacctg tagaatcagc ctttaaactg 3780

ctgtgaactt tgtcatgact tgaatatgaa gatagacaaa aactctgtaa agacaaatgt 3840

ttgttttccc ccttacagaa cgtgtgagct tggttttatc ttcctttgta tttagtcata 3900

acctctcaag ctggcagctc cgaccaagga tcagaagctg tgtgcgttcc acctggtgga 3960

attagctcag ctctatatga gaagtggagt taatggaaaa cgtgttgact gggtggtttc 4020

tatttaaaag agtgatgata attcttgaac agtagttttt attttgctat ttctttaagc 4080

tgactgatgt gccacaaaat tattttaagg tatttgtgtt ttaagagtgt tctcatgaga 4140

ttagttgtag atatttttta aaatacaact ggtttttaaa atctgagtat tgctctaagc 4200

aagtgtttag actcttacgg gaaggtgggt ggaagttgtt tggcttccgt atttccatgc 4260

gtgccgtcag acataggtca gaacgccaac tgtgcatcct gctgtttaaa gacctcttgg 4320

cctctgtgac cctcatgaag gggctgatat tttaagttga ctgtttgatt gtaaattaat 4380

cctttctaat ttttaaagac ttgcttgact gttttccttg ttaaataatt ttaaaaaaat 4440

aaaaaactgg aagttctttg cttaactgta 4470

<210>50

<211>189

<212>PRT

<213> mice

<400>50

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr

20 25 30

Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly

35 40 45

Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr

50 55 60

Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys

65 70 75 80

Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr

85 90 95

Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val

100 105 110

Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys

115 120 125

Gln Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr

130 135 140

Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val

145 150 155 160

Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Asp Asp

165 170 175

Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Leu Met

180 185

<210>51

<211>570

<212>DNA

<213> Guinea pig

<400>51

atgactgagt ataaactggt ggtggttgga gcaggtggtg tcgggaaaag tgcactgacc 60

atccagctaa ttcagaacca ctttgtcgat gaatatgatc ccaccataga ggattcttac 120

cgaaaacagg tggttataga tggtgaaact tgtctgttgg atattctgga tacagctgga 180

caagaggagt acagtgccat gagagaccaa tacatgagga caggcgaagg cttcctctgt 240

gtgtttgcca tcaataatag caaatcattt gcagatatta acctctacag ggagcagatt 300

aaacgagtaa aagactcaga tgatgtacct atggtgctgg tagggaacaa gtgtgatttg 360

ccaacaagga ctgttgacac aaaacaagcc catgaactgg ccaagagtta cgggattcca 420

ttcattgaaa cctcagccaa gaccagacag ggtgttgaag atgcatttta cacactcgta 480

agagaaatac gccagtacag aatgaaaaaa ctcaacagca atgatgatgg gactcaaggt 540

tgtatggggt tgccatgtgt ggtgatgtaa 570

<210>52

<211>189

<212>PRT

<213> Guinea pig

<400>52

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr

20 25 30

Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly

35 40 45

Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr

50 55 60

Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys

65 70 75 80

Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr

85 90 95

Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val

100 105 110

Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys

115 120 125

Gln Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr

130 135 140

Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val

145 150 155 160

Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Asn Asp Asp

165 170 175

Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met

180 185

<210>53

<211>1220

<212>DNA

<213> Guinea pig

<400>53

gttccggggt cctcaacgtt tctcagggtt gagattctat atccttttga agctggggcg 60

gcagagcttg aggttcttgc tggtgtgaaa tgactgagta taaactggtg gtggttggag 120

caggtggtgt cgggaaaagt gcactgacca tccagctaat tcagaaccac tttgtcgatg 180

aatatgatcc caccatagag gattcttacc gaaaacaggt ggttatagat ggtgaaactt 240

gtctgttgga tattctggat acagctggac aagaggagta cagtgccatg agagaccaat 300

acatgaggac aggcgaaggc ttcctctgtg tgtttgccat caataatagc aaatcatttg 360

cagatattaa cctctacagg gagcagatta aacgagtaaa agactcagat gatgtaccta 420

tggtgctggt agggaacaag tgtgatttgc caacaaggac tgttgacaca aaacaagccc 480

atgaactggc caagagttac gggattccat tcattgaaac ctcagccaag accagacagg 540

gtgttgaaga tgcattttac acactcgtaa gagaaatacg ccagtacaga atgaaaaaac 600

tcaacagcaa tgatgatggg actcaaggtt gtatggggtt gccatgtgtg gtgatgtaac 660

aagatattta acaaagttct atcagaaaag agccactttc aagctgcact gataccctgg 720

tcctgacttc cctggaggag aagtatccct gttgctctct tcatctcaga gaagctcctg 780

ctgtttgtcc acctctcagt gtatgagcac agtctctgct tgagaacttc tcagaataac 840

tacctcctca cttggttgtc tgaccagaga aatgcacctc ttgttaattc cccaataatt 900

ttctgccctg ggctctcccc aacaaaaaac aaacacttct gccatccaaa aagcaacttg 960

gtctgaaaca gaaccaaact gtagattgaa attctcttaa aaagtcttga gctctaaagt 1020

tagcaaccgc tggtgatttt tattttcctt tttatttttg aacttggaac tgacctatgt 1080

tagattttgg agaaatgtca taaagtactg ttgtgccaag aagataatta tgttgctgaa 1140

tggttgattt atagtgttat cagctatatt ttacaaactg gcatctgctc tgtattcata 1200

aatacaaaaa tgaagccagg 1220

<210>54

<211>189

<212>PRT

<213> Guinea pig

<400>54

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr

20 25 30

Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly

35 40 45

Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr

50 55 60

Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys

65 70 75 80

Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr

85 90 95

Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val

100 105 110

Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys

115 120 125

Gln Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr

130 135 140

Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val

145 150 155 160

Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Asn Asp Asp

165 170 175

Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met

180 185

<210>55

<211>1307

<212>DNA

<213> dog

<400>55

tgattacgta gcgggcgggg ccggaagtgc cgctccctag tgggggctgt tcatggcggt 60

tccggggtct ccaacctttc tcctagttgt ggtcctaaat acgtcggaag cggaggcggc 120

gaagcttgag gttcttgctg gtgtgaaatg actgagtaca aactggtggt ggttggagca 180

ggtggtgttg ggaaaagcgc actgacaatc cagctaatcc agaaccactt tgtagatgaa 240

tatgatccca ccatagagga ttcttaccga aaacaggtgg ttatagacgg tgaaacctgt 300

ctgttggata tactggatac agctggtcaa gaagagtaca gtgccatgag agaccaatac 360

atgaggacag gcgaaggctt cctctgtgta tttgccatca ataatagcaa atcatttgca 420

gacattaacc tctacaggga acagattaag cgagtaaaag attcagatga tgtacctatg 480

gtgctagtag gaaacaagtg tgatttgcca acaaggacag ttgacacaaa acaagcccat 540

gaactggcca agagttatgg gattccattc attgaaacct cagccaagac cagacagggt 600

gtcgaggatg ccttttacac actggtaaga gaaatacgtc agtaccgaat gaagaaactc 660

aacagcagtg atgatgggac tcaaggttgt atggggttac catgtgtggt gatgtaacaa 720

gacactttta aagttctagc atcagaaaag agccactgtc aagctgcact gacaccctgg 780

tcctgacttc cctggaggag aagtattcct gttgctatct tcagtctcac aaagaagctc 840

ctgctacttc cccaactctc agtagatcag tacaatgttc tctatttgag aagttctccg 900

aacaactacc tcctcacttg gttgtctgac cagagaaatg aacctcttgt tccttcccgc 960

tgtttttcca ccctgaattc tcccccaaca cacataaaca aacctctgcc atcccaggtt 1020

tttcatctga aaaataattc atgctctgaa acagagaaca aaactgtaga catgaaattc 1080

tgtaggaaac aaggtcttga gctcaaaagt agcaactgct ggtgaccttt ttttcccccc 1140

tttttactgt tgaacttgga actatgttgg tttttggaga aatgtcataa gttactgttt 1200

tgctgagaat atagttaagt tgacatttgg tttgtttgta atatcattag ctattttcta 1260

taaattggca tctgctctgc attcataaat acacgagtga attctga 1307

<210>56

<211>189

<212>PRT

<213> dog

<400>56

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr

20 25 30

Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly

35 40 45

Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr

50 55 60

Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys

65 70 75 80

Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr

85 90 95

Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val

100 105 110

Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys

115 120 125

Gln Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr

130 135 140

Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val

145 150 155 160

Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Asp Asp

165 170 175

Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met

180185

<210>57

<211>3104

<212>DNA

<213> Cat

<400>57

aaaaaataaa taaatttaag aaaccatttt aaaattatgc acagttgcag cctggaaaac 60

ttaaggtggc gccttatagt atcaatctta ggagctttat ttggtgcatt taacgcaact 120

ggtaattgca aaatccactt cgcctgtgta agtgaaaaat atagactgtt atcttgttgg 180

ccctatgaaa ttctgcactt ggtatttagc atatactcta ccttcattac tatctggcaa 240

gatgttctgc cttagcactc agttgcattc ttttcctttt ctttcctgtt cattatgctt 300

taattctgag gaccatatga gggtagaata tattaaaaat tacaaaaatt ataaaaattt 360

gtataggcaa accatttcct taagttgatg gccaaatgtt aaaatgttat ttttcatatc 420

atttataatc ttgtcacagt ccacttaacg aagtttggtt agatttcagt gaaaattatc 480

ttccagagta gttttttttt ttttttcctg ggattaggga ggggggtaac tttactgcaa 540

ttagtatgta tggtgcagaa tttcatgcaa atgaggtgtg ccagcagtgt ggtaatttaa 600

tcgtatttaa acaaaaacaa acaaaaaaaa aacgaatgca caaacttgct gctgcttaga 660

tcactgcagc ttctaggacc cagtttcttt tactgatttc aaaacaaaac aaaacaaaaa 720

aataaaaaaa gttgtgcctg aaatgaatct tgtttttttt ataagtagcc gcctggttcc 780

tgtgtcctgt gaaatacagg cacttgaccc ttggtgtagc ttctgttcga ctttatatca 840

cgggaatgga ttggtctgat ttcttggccc tcatcttgaa ttggccacat ccagggtccc 900

tggccagtgg actgaaggct ttgtctaaga ggacaagggc agctcagggg atgtggggga 960

gggcgctttt atcttccccg ttgtcgtttg aggttttgat cttctctggg taaagaggcc 1020

gtttatcttt gtaaacacaa aacatttttg ctttctccag ttttctgtta atggcgaaag 1080

aatggaagcg aataaagttt tactgatttt tgagactcta gcacctagcg ctttcatttt 1140

tgaaacgtcc tgtgtgggag gggcgggtct gggtgcggcc cgccgcgtga ctcctgagtc 1200

gggggcccac gtggctgggg cggggactcg gacgccccgg gcgccgactg attacgtagc 1260

gggcggggcc ggaagtgccg ctccctagtg ggggctgttc atggcggttc cggggtctcc 1320

atcctttttc ccagttgttc taaatcagtc ggaagcggag gcagcgaagt ttgaggttct 1380

cgctggtgtg aaatgactga gtacaaactg gtggtggttg gagcaggtgg tgttgggaaa 1440

agcgcactga caatccagct aatccagaac cactttgtag atgaatatga tcccaccata 1500

gaggattctt accgaaaaca ggtggttata gacggtgaaa cctgtctgtt ggacatactg 1560

gatacagctg gtcaagaaga gtacagtgcc atgagagacc aatacatgag gacaggcgaa 1620

ggcttcctct gtgtatttgc catcaacaat agcaaatcat ttgcagatat taacctttac 1680

agggaacaga ttaagcgagt aaaagactcc gatgatgtac ctatggtgct agtaggaaac 1740

aagtgtgatt tgccaacaag gaccgtcgac acaaaacaag cccacgaact ggccaagagt 1800

tatgggattc cattcattga aacctcagcc aagaccagac agggtgttga agatgccttt 1860

tacacactgg taagagaaat acgtcagtac cgaatgaaga aactcaacag cagtgatgac 1920

gggactcaag gttgtatggg gttaccgtgt gtggtgatgt aacaagatac ttttaaagtt 1980

ctagcatcag aaaagagcca ctgtcaagct gcactgacac cctggtcctg acttccctgg 2040

aggagaagcg ttcctgttgc tattttcagt ttcacaaaga agctcctgct atttccccaa 2100

ctctccgtag atcagtacat tattctctgt ttgagaagtt ctccgaataa ctacctcctc 2160

acttggttgt ctgaccagag aaatgaacct cttgttactc cccactgttt ttccaccctg 2220

gttctccccc agcacatata aacaaacctc ccaggttttt catctgaaaa gtaattcatg 2280

ctctgaaaca gagaaccaaa ctgtagacat gaaattctgt aggaaacaat gtcttgagct 2340

ctaaagtagc aactgctggt gacttttttt tttttttttt cctttttact gttgaacttg 2400

gaactatgtt ggtttttgga gaaatgtcgt aagttactgt tttgctgagt atatagttaa 2460

gtttaccatt cggtttgttt gtaatgtcat tggctatact ctgtacctgg catctgctct 2520

gcattcataa atacaaaagt gaattctgac ttttgagtct atcctagtgt tctcaacttc 2580

cacataatta aatctaactt ttgcagcaaa gtgccttttt cctagaagtg gtttgtagat 2640

ttgctttata atactttggt ggaatagatg tctcaaaaac cattatacat gaaaatgaat 2700

gtctgagata cgtctatgat ctgtctacct ttgagggaaa aatataccga cataatagca 2760

gatgccatgt cttacgtgta tgaagttgga tttccagaga cctgatttgg gtctcttcca 2820

agagaaagat gaaactggaa acaattatga ataacttcac ttaattttta cctaatctct 2880

acttcggggt gggagggcag ggagtaggtt accacttaca aaatatatgc aatttgtttc 2940

ttctagctta ctgataatga acttccattc ttatttaaat ttaggtcata tcctaaagct 3000

ttacatttgc aggtgttcga aattgtaagt ttaatgcagt tttatttaat agctatgatc 3060

aatgattttc aagcctcaga tgtattaacg gacacatttt cact 3104

<210>58

<211>189

<212>PRT

<213> Cat

<400>58

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr

20 25 30

Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly

35 40 45

Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr

50 55 60

Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys

65 70 75 80

Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr

85 90 95

Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val

100 105 110

Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys

115 120 125

Gln Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr

130 135 140

Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val

145 150 155 160

Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Asp Asp

165 170 175

Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met

180 185

<210>59

<211>4283

<212>DNA

<213> cattle

<400>59

ggccgctccc tagtgggggc tgttcatggc ggttccgggg tctcccaaca attttcccgg 60

ttgtggtcgt aatctatccg aagtggaggc agtggagcta gaggttcttg ctggtgtgaa 120

atgactgagt acaaactggt ggtggttgga gcaggtggtg ttgggaaaag tgcactgaca 180

atccagctaa tccagaacca ctttgtagat gaatatgatc ccaccataga ggattcctac 240

cgaaaacagg tggttataga tggtgaaacc tgtctgttgg acatactgga tacagctgga 300

caagaggagt acagtgccat gagagaccaa tacatgagga caggcgaagg cttcctttgt 360

gtgtttgcca tcaataatag caaatcattt gcagatatta acctctacag ggaacagata 420

aagcgtgtaa aggactcgga tgatgtacct atggtgctag taggaaacaa gtgtgatttg 480

ccaacaagga cagttgacac aaaacaagcc catgaactgg ccaaaagtta tgggattcca 540

ttcattgaaa cctcagccaa gaccagacag ggtgttgaag atgcctttta cacactggta 600

agagaaatac gtcagtaccg aatgaaaaag ctcaacagca gtgatgatgg cactcaaggc 660

tgtatggggt tgccgtgtgt ggtgatgtaa caagatactt ttaaagttct cacatcagaa 720

aagagccact gtcaagctgc actgacaccc tggtcctgac ttccctggag gagaagtatt 780

cctgttgcta tcttcagttt caaaaagaag ctcctgctat ttccccaact ctcagtagat 840

caatataata ttctctattt gagaagttct caagaataac tacctcctca cttggttgtc 900

tgaccagaga attgaacctc ttgttactcc cagtattttt ccaccctggg ttctccccca 960

gcacacacaa acgcacctct gccacccagg tttttcatct gaaaagcaat taatactctg 1020

aaacagagaa ccaaactgta gaaacatgaa attctgtaga aaacaatgtc ttgagctcta 1080

aagtagcaac tgctggtgat tttttttttt tttttttcct ttttattgtt gaacttggaa 1140

ctatgttggt ttgtggagaa atgtcataaa ttactgtttt gctgagaata tagttaatgt 1200

tgctctctgg tttgtttgta atgttatcag ctatattcta taaactggca tctactctgt 1260

atttagaaat acaaaaatga atactgacct tttgagtcta ccctcatctt ctcgactttc 1320

ttgtaattaa atgtaacttt cacgatgaag tgccttttgc ctgggagtga ctcgtagact 1380

tcctttaaaa tacttcagtg gaatagatgt ctcagaaact gttatacata agaataaatg 1440

tctgagatat gtctatgacc catctagctt tgagggaaag atataccaat atgatagcag 1500

atgccatttc ttacatctat aacgttgatt ttctggagac ctattttggg gctctccgag 1560

agaaagatga gactataaat gattaggaat aatttcactt aatttttaca taacctccac 1620

tttttgtttt gtagtttact acctgcaaaa catataattt gattcctttt agcttacaga 1680

taatctaatg ttaaatgaac ttccattcat attttaattt ggatcatatc aggaagtcta 1740

catttgcagg tgttcaaaaa ttgtaaaagt gtgatgcagt tttatttaat agctgtgatc 1800

aatgattttc aagcctcaaa tatgttaata gacacatttt cactgtatat catggtatta 1860

ataattattg atgtatataa ttgtccttgg tccccttctc tgttcatcac ctcatggcaa 1920

tggcttgatt aattatttca gctgagtaaa gcatggtgct aatagaccag ggtcacagtg 1980

tcaaaacttc agtgagccag taagcatcac agagaaagaa attctttcac atttgctcac 2040

cattaactcc agctaatagt tttgccagat gtgtgtggtt agtcctgcaa ggaaaggaga 2100

agtcagttaa tacaaattct taaccaggac tggaaaaact tgttttcctg agaagggtca 2160

gcttagaagt ctttatctgg actctatttt tagccacatg gaaatcaaat taagctgatc 2220

ttttttctca agtttttgag agtgaggatg cctcagatca acatttttaa aatattcttt 2280

attcttacgt tcttttaagg gtttaaaaca acgttgagta attagtctgg gcataccagg 2340

taacaagctg ataagtttgt gctgaacaag aagtagcctt tggattgaaa ttgctgtttt 2400

gagaagggat agaaaatata attaataatt atgagacttg acttttctat ttgcagataa 2460

tatcctgata attctgatga aaatagactt ggataatttt tgataaaaga atcgttccaa 2520

aatggccact tgctgttctt gtcttctaat gtgtaaatac ttactgaggt cctcttctaa 2580

tatgagttgt catttattaa gcaaattcca cattgccttg aaatgaattc ggaagagaag 2640

aaaaagtcat agtataccca gagaatgaaa aatccagaga attgtgctcc ttagtgttaa 2700

ttctgaagcc ttcgtagtcc acacccatag acagaaactc tctgccactt tgcttctgct 2760

cctcttggag cattgcgctg tcatttcctt gaggatagat tgaggcttgt caactcagtt 2820

gtattgtctt cctcctcttc ctcttgtctg tgtgactgac agtgtgactc ttactaatgt 2880

cagatgcggg gatgcgggga ggtggggggg agtagctcat tttaggctct tgcacccttt 2940

accgttgtat gtgtgtgtct tttagttttc tcaagaatgt tctaagcaca gaagtatcta 3000

aatggggcca aaattcagac ttgaaaatgt tcttttaata gcttcttaaa aagttacact 3060

ttggtgtgaa ttttggcagg atagagtgac aaactcttaa acgctgaata acttcagtta 3120

gtgtgttata gtttttagaa tatgtttgtg attgctgaaa acaattatag tttacctcaa 3180

aatctgaaag tctctttccc caagttaagt gcctggccag ctgtcaaaga ttacatatta 3240

ctttatgttt gtttgttttt taaaggttgc acattcaaga ttgtgaaaat aaggtgttct 3300

gtctgaaagc taccatgcct gtctgtaaat gaatccactg agtgctgtac ttgttccaac 3360

agcttactac agaatgctac ttggtaatat catactcgtt acagttttca cttcaggagt 3420

gtactaggta gaatgatcct gtgtgtattg tagtgggctc catgtttagt cttttcagca 3480

tcctttaaac tgctgtgaat ttttgtcttg acttgaaagc aaggatagag aaacacttta 3540

aagagatact ttgggttttt ttccattcca gaattggtga gcatagttag attttgcttt 3600

acatttacag tcatgaactc ttaagctggc agctacaacc aagaaccaaa agagggtgca 3660

ttctgcttct tgtaattcat ctttgctaat aaattatgag aagcaaagat aattaattag 3720

agaaactatt ttatttgggt ggtttctata aacaagggac tataattctt aaacattatt 3780

tttcattttt gctgtttctt taagaaacct aatgtgccac aacattattt taaggtgttt 3840

cttaaaagaa ttgtttttaa aagtgttctc attttcagag taattgtaga tatatttcaa 3900

aatataactg ataattttta aaggcctgag tactgaccta agaagcagtt gtatgaattc 3960

tctgggggga agggaggagc tcagtgaaag ttgtatgact tttatatttc tgtgccatca 4020

aataaaggta aaaatgtctt ttgtgcagtt ttgctgttca aacagaaact attggcctcc 4080

ttggccctaa atgaaagggc tggtatttta agttgactat tttattgtaa attaatccat 4140

cttaattttt ttaaatttgg ttgaatgttc tcttgttaaa tgtttaaaaa ataaaaactg 4200

gaagttcttt gcttagtcat aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 4260

aaaaaaataa aaaaaaaaaa aaa 4283

<210>60

<211>189

<212>PRT

<213> cattle

<400>60

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr

2025 30

Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly

35 40 45

Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr

50 55 60

Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys

65 70 75 80

Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr

85 90 95

Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val

100 105 110

Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys

115 120 125

Gln Ala His Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr

130 135 140

Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val

145 150 155 160

Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Ser Asp Asp

165 170 175

Gly Thr Gln Gly Cys Met Gly Leu Pro Cys Val Val Met

180 185

<210>61

<211>4825

<212>DNA

<213> Chicken (Gallus Gallus)

<400>61

gcgccgggac cggaagccgg aagctttgca gaagggtgtt ccgcgttcgc ggtgcgggag 60

cggtcagccg gggtggcggg gctggggccg gccggggcag gcggctccgc gctccgcact 120

gggccgctgg gagggcgatg actgaataca agctggtggt ggtgggagct ggcggcgtcg 180

ggaagagcgc gttgaccatc cagctcatcc agaaccactt cgtggacgag tacgacccca 240

ccatcgagga ttcgtacaga aagcaggttg tcatcgatgg agagacgtgc ttgttggaca 300

ttctggacac tgcaggacag gaagaataca gtgctatgcg tgatcagtac atgagaactg 360

gggaaggatt cctttgtgtg tttgccatta acaacagtaa atcattcgct gatattaacc 420

tttacagaga gcaaatcaag agagtgaaag attcagacga tgtgccaatg gtgctggtgg 480

ggaataagtg cgatttgcca acaaggacag tagacaccaa acaggctcaa gagttagcaa 540

aaagctacgg cattcccttc atagagacat cagccaaaac gagacagggt gtggaagatg 600

cgttttacac actggtgagg gagattcggc agtaccggat gaaaaagctc aacagcaacg 660

aagatgggaa tcagggctgt atggggttgt cctgcattgt gatgtgataa gatgccaggt 720

tcagatgtag ctgctggaca agtctcgatg ctactgtatt gtgtctcatg ctgatgccct 780

gcagtatttt ggtgccagcg accagactct tggtaccagt taattagctc aggatccttt 840

cctgtgctcc atctgaagaa aacatctctg gtatctacct ccttgctcag ctcacagagc 900

agtcatatct cttggtgtac tgggattctt ttctagctgt gttgtctggg tttgttcaag 960

aagaaaacca gtcacaagaa aagtgaatta cagagactaa atgctgtgaa aaagatcaca 1020

ctttacctcc agagtaaaag ctagaagtgg cgtttgaccc ctttgcattg gattcagatt 1080

tgcggtgttg tcagaggagt ggcagaagta attttgccat tacaaaggtt tctgtcacca 1140

gtcggattgg tatctgctgt ctgtgcaccc acacagtgta tctgcaacat ctgcattgtg 1200

ccagaagtat cacttaactg atgaactgat cctttatttt tctgtaataa aaaggagata 1260

tctttgctaa cttaagtgcc tgtttgctca gaaggttgga ggttgtatgc tgttcccttg 1320

ggctgaggag aaccccaagg atgaatttct tgggtgctca ttgtcttgag caggcaagtt 1380

ttgtgtgggt gatctctttt catggcagga tattaaaatg ggaatttgta gtctggaaga 1440

tggagcagct gtttgtgaga ctcttgagtt agggagagaa atgtatacca cgtctgttct 1500

cgatccatca gaatggatcc atccacctct ttgtgtgtgg aactgtgtat agtctgtatt 1560

ggttttctac agcacttgga tctctttgga ccaaattagc gagctgttca ttttaacata 1620

actgccagta tttatagaca atttcttacg gacagataat gaatttagaa actggaggtt 1680

actttgggca gctgttcctc agctctgtct gtaacttgca aattattctg agttattttc 1740

tgcagaacct ccttccttat cacgggagga gcctgggagt tgaggttgac tgtaattggg 1800

tcaatggttg tcacagactt aaggtgtcca ggctgattgg aggaggcact gagccctaac 1860

agagcactga gctgacttct aattgcagca tccttgcaaa atgaggaagg gagttcagtg 1920

atgtctgcac tgaagatgta tgatacactg atagcagttc tgggtatgtt gtaacagctt 1980

caaagtagaa ccgcagtact gcgtgagctg tgtgacttct tcctagaaca cagcactgtc 2040

accccatatg gttgggacgt gcaggtgaga ccaacaccta ccaggttccc tggcgtaccg 2100

tggccttctc agttcttgtg ccagtgatac tgggttctgt tctgtggtgt cagacagcgt 2160

cctgtagcaa agctgaattc ccacttagtc tggtgagaga ataaagagcc atcagccaac 2220

agagggagcg ttcattctgc tggagcagtg cgagctgtaa gcattacgag aggcgtagtt 2280

tcagtttgtt gcagtcaggt tcctatattt tcaaagctga aatcagaaat aagtaaatac 2340

ggagaaaata agctgttgct tttaatgctc tttcctccac taattgtact cttaattttc 2400

ttcttgggag gccgaggatc catctgcata actttagctg tgatgctcca gataagtgtt 2460

tagaattcat tttatctttg actgatggga ctgataagaa gttaacgcac aatattttta 2520

catacaacat cgttttccag tgacctcctg agcggtggga agcattatgg gatagcaccg 2580

gctgtgactc gagttcattt gaaggcgatc tcttgcctgc aggttaaatg ggacggagtc 2640

agaatcactg tgagccgtct gtaatcagca aacagtctgt gggcttttct tactgtgttc 2700

tctctgtttg ccttagtttg gtgcaggaag agttccttgt gacagcgtcc tttgaggtgt 2760

gttgcaggag ctgaccattt gctccttgag ctgtgtgatg aactgttgtc cacttaatgg 2820

agttacagaa gcagcttctg ggagtcgcat ctggtcgcat acattcagtg ttttgggaag 2880

ctgtcagtgt ggtgtttgca ctgtgtttga atggtgttca tggtgggtct gttatgctcc 2940

tggatgattt ggggagatgt ggggctgctt ccgtggcaga caggatcagc tcagggcgct 3000

gctgcctatg gctgtgggaa acctcacagt tggtgtttga atagtggcca agtatgtcaa 3060

ttaaaaatac attttgaagg gaggtttgtc atagctctgt actttggcat gctctgctta 3120

ctgaaaacat actagctgta gctcaaaaaa agttgtgaat cctcagaata atacaggagc 3180

tggcaattgt ggctgctttc tctttgtgtt ccttttctct tgggttggat gaagctttaa 3240

aaaggaagga gccctggtga gggttggtca gtgtgcattt cattcttgga accagagagg 3300

aagttgcatc aactttcagg acgctgcaga gctcacttgc acaggtggtg ctccagtcta 3360

tgtgattttt ggggtcaaat cttgagatga tcttacaaaa tcagattttg tacccatcat 3420

gagcatgagg tgagtggttg tgctcggttt ctagctgcat gtatgtatac agacacgtgt 3480

atgcagacat gtctatgtgt gagtagttcg agtcagtcaa ggttactggc agcacctaaa 3540

gcgtatgcac cacataatgc atgcaggcaa aagtcctatc ttaggagcca tctcttcatg 3600

ggtttgggtt tatataggca gtatttttaa acagaatatc cgaagcactt tctggagttc 3660

tgtggtaatg cagtgacacc tatttggatg aaggaagatg tgtctgagga gcacgtaagc 3720

agatttgctg ccctaacaga gaggttttgg taaccgtgga aaaggttttc tcctggatct 3780

gtgtgtgctc ttggtgagct gcaatccatg acagggcaca accagatgag aaggaaaccc 3840

ggccatccca tgcttgagca cagctctgac tcagtagttc caccagatgt gccctttcag 3900

tcaaagtgtt ctgatctctt agagctttct gtagttcaag ttaccactca ctctccagct 3960

tgctcggtta atgtctgttg gcggcgttga gttggacttg ggaaaggtgt gtgtggtagg 4020

aacaagcaga gtgtgatgtg cttctgttat caggacttaa gctagagtgg ttggcagata 4080

ggaaatgcag ctattccttg aaagcaagca gatcatggat ggtcagccaa actgccctgg 4140

ctttggtggg agctgcactg cagaaggacc aaaccccaac aagatttggc acatttgttt 4200

agaagataag cacagatggt tttgcacaag gcagctcctc ataatggtgg ctttgtagat 4260

ttagtccaaa tgttcttatt tagatctagc agcacatcac tgtgtccgtg cccatctaac 4320

ctcgctatcc taagtagagc agaccccaaa caaccttgtt caaaaactac cagtgcaaat 4380

aactgaacta aatatttgtt actgctgact gagaacagct gttcgagtgt agcattgtgg 4440

cttgttaatg tgagtgcccc aactctatgg tcttattaaa gaaacccaaa cattgctcag 4500

attttgttct tattgtcatc ataagacttg aatagtgatg gtaatgctta cgtagacgtg 4560

tcttgtgagt gcacttcagt gatttagaaa gaactggatt tcaagcaact ttggacctgt 4620

ggggggaggg agattaatga aggtttgaat cacattctaa ttctatgtac agtccttcat 4680

tactccacaa gcctaaatcc tatacagcct ccaggatagc tggaaactgt tgagatctgg 4740

actttttttt tttaatccaa gggctaactt gttgtaactt ggtataatta tctgctttcg 4800

gaaatgcatc tctgttggtt tgaaa 4825

<210>62

<211>189

<212>PRT

<213> Chicken

<400>62

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr

20 25 30

Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly

35 40 45

Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr

50 55 60

Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys

65 70 75 80

Val Phe Ala Ile Asn Asn Ser Lys Ser Phe Ala Asp Ile Asn Leu Tyr

85 90 95

Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Asp Asp Val Pro Met Val

100 105 110

Leu Val Gly Asn Lys Cys Asp Leu Pro Thr Arg Thr Val Asp Thr Lys

115 120 125

Gln Ala Gln Glu Leu Ala Lys Ser Tyr Gly Ile Pro Phe Ile Glu Thr

130 135 140

Ser Ala Lys Thr Arg Gln Gly Val Glu Asp Ala Phe Tyr Thr Leu Val

145 150 155 160

Arg Glu Ile Arg Gln Tyr Arg Met Lys Lys Leu Asn Ser Asn Glu Asp

165 170 175

Gly Asn Gln Gly Cys Met Gly Leu Ser Cys Ile Val Met

180 185

<210>63

<211>2603

<212>DNA

<213> human

<400>63

aggcgaggct tccccttccc cgcccctccc ccggcctcca gtccctccca gggccgcttc 60

gcagagcggc taggagcacg gcggcggcgg cactttcccc ggcaggagct ggagctgggc 120

tctggtgcgc gcgcggctgt gccgcccgag ccggagggac tggttggttg agagagagag 180

aggaagggaa tcccgggctg ccgaaccgca cgttcagccc gctccgctcc tgcagggcag 240

cctttcggct ctctgcgcgc gaagccgagt cccgggcggg tggggcgggg gtccactgag 300

accgctaccg gcccctcggc gctgacggga ccgcgcgggg cgcacccgct gaaggcagcc 360

ccggggcccg cggcccggac ttggtcctgc gcagcgggcg cggggcagcg cagcgggagg 420

aagcgagagg tgctgccctc cccccggagt tggaagcgcg ttacccgggt ccaaaatgcc 480

caagaagaag ccgacgccca tccagctgaa cccggccccc gacggctctg cagttaacgg 540

gaccagctct gcggagacca acttggaggc cttgcagaag aagctggagg agctagagct 600

tgatgagcag cagcgaaagc gccttgaggc ctttcttacc cagaagcaga aggtgggaga 660

actgaaggat gacgactttg agaagatcag tgagctgggg gctggcaatg gcggtgtggt 720

gttcaaggtc tcccacaagc cttctggcct ggtcatggcc agaaagctaa ttcatctgga 780

gatcaaaccc gcaatccgga accagatcat aagggagctg caggttctgc atgagtgcaa 840

ctctccgtac atcgtgggct tctatggtgc gttctacagc gatggcgaga tcagtatctg 900

catggagcac atggatggag gttctctgga tcaagtcctg aagaaagctg gaagaattcc 960

tgaacaaatt ttaggaaaag ttagcattgc tgtaataaaa ggcctgacat atctgaggga 1020

gaagcacaag atcatgcaca gagatgtcaa gccctccaac atcctagtca actcccgtgg 1080

ggagatcaag ctctgtgact ttggggtcag cgggcagctc atcgactcca tggccaactc 1140

cttcgtgggc acaaggtcct acatgtcgcc agaaagactc caggggactc attactctgt 1200

gcagtcagac atctggagca tgggactgtc tctggtagag atggcggttg ggaggtatcc 1260

catccctcct ccagatgcca aggagctgga gctgatgttt gggtgccagg tggaaggaga 1320

tgcggctgag accccaccca ggccaaggac ccccgggagg ccccttagct catacggaat 1380

ggacagccga cctcccatgg caatttttga gttgttggat tacatagtca acgagcctcc 1440

tccaaaactg cccagtggag tgttcagtct ggaatttcaa gattttgtga ataaatgctt 1500

aataaaaaac cccgcagaga gagcagattt gaagcaactc atggttcatg cttttatcaa 1560

gagatctgat gctgaggaag tggattttgc aggttggctc tgctccacca tcggccttaa 1620

ccagcccagc acaccaaccc atgctgctgg cgtctaagtg tttgggaagc aacaaagagc 1680

gagtcccctg cccggtggtt tgccatgtcg cttttgggcc tccttcccat gcctgtctct 1740

gttcagatgt gcatttcacc tgtgacaaag gatgaagaac acagcatgtg ccaagattct 1800

actcttgtca tttttaatat tactgtcttt attcttatta ctattattgt tcccctaagt 1860

ggattggctt tgtgcttggg gctatttgtg tgtatgctga tgatcaaaac ctgtgccagg 1920

ctgaattaca gtgaaatttt ggtgaatgtg ggtagtcatt cttacaattg cactgctgtt 1980

cctgctccat gactggctgt ctgcctgtat tttcgggatt ctttgacatt tggtggtact 2040

ttattcttgc tgggcatact ttctctctag gagggagcct tgtgagatcc ttcacaggca 2100

gtgcatgtga agcatgcttt gctgctatga aaatgagcat cagagagtgt acatcatgtt 2160

attttattat tattatttgc ttttcatgta gaactcagca gttgacatcc aaatctagcc 2220

agagcccttc actgccatga tagctggggc ttcaccagtc tgtctactgt ggtgatctgt 2280

agacttctgg ttgtatttct atatttattt tcagtatact gtgtgggata cttagtggta 2340

tgtctcttta agttttgatt aatgtttctt aaatggaatt attttgaatg tcacaaattg 2400

atcaagatat taaaatgtcg gatttatctt tccccatatc caagtaccaa tgctgttgta 2460

aacaacgtgt atagtgccta aaattgtatg aaaatccttt taaccatttt aacctagatg 2520

tttaacaaat ctaatctctt attctaataa atatactatg aaataaaaaa aaaaggatga 2580

aagctaaaaa aaaaaaaaaa aaa 2603

<210>64

<211>393

<212>PRT

<213> human

<400>64

Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp

1 5 10 15

Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala

20 25 30

Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys

35 40 45

Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys

50 55 60

Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly

65 70 75 80

Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Val Met Ala Arg

85 90 95

Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile

100 105 110

Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly

115 120 125

Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu

130 135 140

His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg

145 150 155 160

Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly

165 170 175

Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys

180 185 190

Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp

195 200 205

Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val

210 215 220

Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr

225 230 235 240

Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met

245 250 255

Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu

260 265 270

Leu Met Phe Gly Cys Gln Val Glu Gly Asp Ala Ala Glu Thr Pro Pro

275 280 285

Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser

290 295 300

Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu

305 310 315 320

Pro Pro Pro Lys Leu Pro Ser Gly Val Phe Ser Leu Glu Phe Gln Asp

325 330 335

Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu

340 345 350

Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu

355 360 365

Val Asp Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro

370 375 380

Ser Thr Pro Thr His Ala Ala Gly Val

385 390

<210>65

<211>2151

<212>DNA

<213> rat

<400>65

cggccgcgcg ctccctgctg agttgcaggc tgtttcccgg ctgcaagatg cccaagaaga 60

agccgacgcc catccagctg aacccggccc ccgatggctc cgcggttaac gggaccagct 120

cggccgagac caacctggag gccttgcaga agaagctgga ggagctggag ctggacgagc 180

agcagcggaa gcgccttgag gcctttctga cgcagaagca gaaggtggga gagttgaagg 240

atgatgactt tgagaagatc agtgaactgg gggctggcaa tggtggagtg gtgttcaagg 300

tctcccacaa gccatctggc ctggttatgg ctaggaagct aattcacctg gagatcaaac 360

ccgcaatccg gaaccagatc atccgggagc tgcaggtgct gcatgagtgc aactccccgt 420

acatagtggg cttctacggg gccttctaca gtgacggcga gatcagcatc tgcatggagc 480

acatggatgg tgggtccttg gatcaagtgc tgaagaaagc tggaagaatt cctgagcaaa 540

ttttaggaaa agtcagcatc gctgtgataa aaggcctgac atatctacga gagaagcaca 600

agattatgca cagagatgtc aagccttcca acattctagt gaactcacgt ggggagatca 660

aactctgcga ttttggggtc agcgggcagc taattgactc catggccaac tccttcgtgg 720

gaacaaggtc ctacatgtcg cctgagagac tccaggggac tcactactct gtgcagtcgg 780

acatctggag catggggctc tctctggtgg agatggcagt tggaagatac cccattcctc 840

ctcctgatgc caaggagctg gagctgctgt ttggatgcca ggtggaagga gacgcggccg 900

aaacgccacc caggccaagg acccctggga ggcccctcag ctcatatgga atggatagcc 960

gacctcccat ggcaattttt gagttgttgg attacatcgt caatgagcct cctccaaaac 1020

tgcccagtgg agtattcagt ctggaatttc aggattttgt gaataagtgc ttaataaaga 1080

accctgcaga gagagcagat ctgaagcagc tcatggtaca tgctttcatc aagagatctg 1140

atgccgagga ggtagacttc gcaggctggc tctgctccac cattgggctt aaccagccca 1200

gcacaccaac ccacgctgcc agcatctgag cctttgggaa gcagcagaga ggaatcctct 1260

gcccagtggc atgccatgtt gctttcaggc ctctcccatg cttgtctatg ttcagatgtg 1320

catctcatct gtgacaaagg atgaagaaca cagcatgtgc caaatcgtac ttgtgtcatt 1380

tttaatattg tctttatcgc tatggttact cccctaagtg gattggcttt gtgcttgggg 1440

ctatttgtct gttcatcaaa tacatgccag gttgaactac agtgaaaccc tggtgacctg 1500

ggtggtcttc ttactgatgt ttgcgctgct gttcatcgtg actcactagc tggctgcctg 1560

tattgtcagg attctcggac ccttggtact tcactcttgc tggtgacctc tcagtctgag 1620

gagagggggc cttctgagac ccttcacagg cagtgcatgc atgaaaagca tgctttgctg 1680

ctactgaaat gagcaccaga acgtgtacat catggtattt tatttttgct tttggtatag 1740

aactcagcag ttcccattta aaaaaaaaat ctaaccagag cccatcactg ccatgatagc 1800

tggggcttca gtctgtctac tgtggtgatt tttagacttc tggttgtatt tctatattta 1860

tttttaaata tactgtgtgg gatacttagt ggtatatgtc tctgagtttg gattagtgtt 1920

tctaaattgg tagttatttt gaatgtcaca aatggattaa ggaatcaacg tatcaagagt 1980

tctatctttc ttccagtcta agtaccaatg ctattgtaaa cgtgtatagt gcctacaaat 2040

tgtatgaaaa cccttttaac cactttactc aagatgttta tcaaatctaa tctcttattc 2100

taataaaaat actatcaagt taaagtaaaa aaaaaaaaaa aaaaaaaaaa a 2151

<210>66

<211>393

<212>PRT

<213> rat

<400>66

Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp

1 5 10 15

Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala

20 25 30

Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys

35 40 45

Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys

50 55 60

Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly

65 70 75 80

Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Val Met Ala Arg

85 90 95

Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile

100 105 110

Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly

115 120 125

Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu

130 135 140

His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg

145 150 155 160

Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly

165 170 175

Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys

180 185 190

Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp

195 200 205

Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val

210 215 220

Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr

225 230 235 240

Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met

245 250 255

Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu

260 265 270

Leu Leu Phe Gly Cys Gln Val Glu Gly Asp Ala Ala Glu Thr Pro Pro

275 280 285

Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser

290 295 300

Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu

305 310 315 320

Pro Pro Pro Lys Leu Pro Ser Gly Val Phe Ser Leu Glu Phe Gln Asp

325 330 335

Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu

340 345 350

Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu

355 360 365

Val Asp Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro

370 375 380

Ser Thr Pro Thr His Ala Ala Ser Ile

385 390

<210>67

<211>2387

<212>DNA

<213> mice

<400>67

agtccctcac tgggacgtct gtgcgcggcg tctcggagcg ccggagcagc ggtggccgca 60

ctttctccaa gctggggctg tagctgagct gtgggtagtg cgcagggagc cgtccgagcc 120

cgaggaaccg gtgtgctgag gcgagagttc ccggccggcg agcgcgcgca gctggttctc 180

cgcgtgggtt gggcggaggg tcccaggagc gcggcgttga tcgagccgcc ccgactctgg 240

gcagagccga gggaggaagc gagaagcggc cgcgcgctcc ctgctgagtt gcaggctctt 300

tcccggctgc aagatgccca agaagaagcc gacgcccatc cagctgaacc cggcccccga 360

tggctcggcg gttaacggga ccagctcggc cgagaccaac ctggaggcct tgcagaagaa 420

gctggaggag ctggagcttg acgagcagca gcggaagcgg ctcgaggcct ttctgacgca 480

gaagcagaag gtgggggaac tgaaggatga tgactttgag aagatcagcg aactgggagc 540

tggcaacggt ggagtggtct tcaaggtctc ccacaagcca tctggcctgg ttatggctag 600

aaagctgatc cacctggaga tcaaacccgc aatccggaac cagatcatcc gggagctgca 660

ggtactgcac gagtgcaact ccccgtacat cgtgggcttc tacggggcct tctacagcga 720

cggcgagatc agcatctgca tggagcacat ggatggtggg tccttggatc aagttctgaa 780

gaaagctgga agaattcctg agcaaatttt aggaaaagtt agcattgctg tgataaaagg 840

cctgacctat cttcgggaga agcacaagat tatgcacaga gatgtcaagc catccaacat 900

tctagtgaac tcacgtgggg agatcaaact ctgtgatttt ggggtcagcg ggcagctaat 960

tgactctatg gccaactcct tcgtgggcac gagatcctac atgtcgcctg agagactcca 1020

ggggactcac tactctgtgc agtcggacat ctggagcatg gggctctctc tggtggagat 1080

ggcagttggg agatacccca ttcctcctcc tgatgccaag gagctggagc tactgtttgg 1140

atgccatgtg gaaggagacg cagccgaaac accacccagg ccaaggaccc ctgggaggcc 1200

tctcagctca tatggaatgg acagccgacc tcccatggca atttttgagt tgttggatta 1260

cattgtcaat gagcctcctc caaaactgcc cagtggagta ttcagtctgg agtttcagga 1320

ttttgtgaat aaatgcttaa taaagaaccc tgcagagaga gcagatctga agcagctcat 1380

ggtacatgct ttcatcaaaa gatctgacgc cgaggaggta gacttcgcag gctggctctg 1440

ctccaccatt gggcttaacc agcccagcac accaacccac gctgccagca tctgagcctt 1500

taggaagcag caaagaggaa ttctctgccc agtggcatgc catgttgctt tcaggcctct1560

cccatgcttg tctatgttca gacgtgcatc tcatctgtga caaaggatga agaacacagc 1620

atgtgccaaa ttgtacttgt gtcattttta atatcattgt ctttatcact atggttactc 1680

ccctaagtgg attggctttg tgcttggggc tatttgtctg ttcatcaaac acatgccagg 1740

ctgaactaca gtgaaaccct agtgacctgg gtggtcgttc ttactgatgt ttgcactgct 1800

gttcatcgtg actcactagc tggctgcctg tattgtcagg attctcggac cttggtactt 1860

cactcttgct ggtgacctct cagtctgaga gggagccttg tgagaccctt cacaggcagt 1920

gcatgcatgg aaagcatgct ttgctgctac tgaaatgagc atcagaacgt gtacgtcatg 1980

gtatttttat tttttgcttt tggtatagaa ctcagcaatt cccatcaaaa aaacctaagc 2040

agagcccatc actgccatga tagctgggct tcagtctgtc tactgtggtg atttttagac 2100

ttctggttgt atttctatat ttatttttaa atatacagtg tgggatactt agtggtgtgt 2160

gtctctaagt ttggattagt gtttctaaat tggtggttat tttgaatgtc acaaatggat 2220

taaagcatca atgtatcaag agttctatct ttcttccagt ctaagtacca atgctattgt 2280

aaacaacgtg tatagtgcct acaaattgta tgaaacccct tttaaccact ttaatcaaga 2340

tgtttatcaa atctaatctc ttattctaat aaaaatacta tcaagtt 2387

<210>68

<211>393

<212>PRT

<213> mice

<400>68

Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp

1 5 10 15

Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala

20 25 30

Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys

35 40 45

Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys

50 55 60

Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly

65 70 75 80

Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Val Met Ala Arg

85 90 95

Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile

100 105 110

Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly

115 120 125

Phe Tyr Gly Ala PheTyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu

130 135 140

His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg

145 150 155 160

Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly

165 170 175

Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys

180 185 190

Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp

195 200 205

Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val

210 215 220

Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr

225 230 235 240

Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met

245 250 255

Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu

260 265 270

Leu Leu Phe Gly Cys His Val Glu Gly Asp Ala Ala Glu Thr Pro Pro

275 280 285

Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser

290 295 300

Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu

305 310 315 320

Pro Pro Pro Lys Leu Pro Ser Gly Val Phe Ser Leu Glu Phe Gln Asp

325 330 335

Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu

340 345 350

Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu

355 360 365

Val Asp Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro

370 375 380

Ser Thr Pro Thr His Ala Ala Ser Ile

385 390

<210>69

<211>1182

<212>DNA

<213> Rabbit

<400>69

atgccaaaga agaagcccac ccccatccag ctgaatcctg cccctgacgg ctcggcggtg 60

aatggtacca gctcggcgga gaccaacctg gaggccttgc agaagaagct ggaggagctg 120

gagcttgacg agcagcagcg gaagcgcctg gaggccttcc tcacccagaa gcagaaagtg 180

ggagagctga aggacgatga cttcgagaag atcagtgagc tgggagccgg caacggcggc 240

gtggtgttca aggtctccca caagcccagt ggcctggtga tggccagaaa gcttattcac 300

ctggagatca aacctgctat ccggaaccag atcataaggg agctgcaggt tctgcacgag 360

tgcaactccc cgtacatcgt gggcttctac ggggcattct acagcgatgg cgagatcagc 420

atctgcatgg agcacatgga cgggggttcc ttggatcaag tcctgaagaa agctggacgg 480

attcccgagc aaattttggg gaaagttagc attgctgtga tcaagggcct gacgtatctg 540

agggagaagc acaagatcat gcacagagat gtgaagccct ccaacatcct ggtcaactcc 600

cgcggggaga tcaagctctg tgacttcggg gtcagtgggc agctcatcga ctccatggcc 660

aactccttcg tgggcaccag gtcttatatg tcgcccgaga gactccaggg gacacactac 720

tctgtgcagt cggacatctg gagcatgggg ctgtccctgg tggagatggc ggtggggcgg 780

taccccatcc cgccccccga cgccaaggag ctggagctga tgtttgggtg ccaggtggag 840

ggcgatgcgg ccgagactcc gcccaggccc aggacccctg ggcggcccct cagctcgtat 900

ggaatggata gccggcctcc catggcgatt tttgagctgc tggattacat cgtcaatgag 960

cctcctccga aactccccag cgcagtcttc agcctggagt ttcaagattt tgtgaataaa 1020

tgcttaataa aaaaccccgc cgagagagca gacttgaagc agctcatggt tcatgctttt 1080

atcaagaggt ctgatgccga ggaggtggat tttgctggtt ggctgtgctc caccatcggc 1140

cttaaccagc ccagcacgcc gacgcacgcg gccggtgtgt ga 1182

<210>70

<211>393

<212>PRT

<213> Rabbit

<400>70

Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp

1 5 10 15

Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala

20 25 30

Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys

35 40 45

Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys

50 55 60

Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly

6570 75 80

Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Val Met Ala Arg

85 90 95

Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile

100 105 110

Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly

115 120 125

Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu

130 135 140

His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg

145 150 155 160

Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly

165 170 175

Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys

180 185 190

Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp

195 200 205

Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val

210 215 220

Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr

225 230 235 240

Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met

245 250 255

Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu

260 265 270

Leu Met Phe Gly Cys Gln Val Glu Gly Asp Ala Ala Glu Thr Pro Pro

275 280 285

Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser

290 295 300

Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu

305 310 315 320

Pro Pro Pro Lys Leu Pro Ser Ala Val Phe Ser Leu Glu Phe Gln Asp

325 330 335

Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu

340 345 350

Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu

355 360 365

Val Asp Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro

370 375 380

Ser Thr Pro Thr His Ala Ala Gly Val

385 390

<210>71

<211>1872

<212>DNA

<213> Guinea pig

<400>71

cgtgtcttcg tcgggaccgc cctcctcctt gagtcctccc cccaccggga cggccgagtg 60

gagaggccgg acgaaggcgg cggccccggc ggcggctttt cctcggcttc gctgtgcagc 120

gtgcgcggcg aggttgaccg cccgcgagcg cccgtgactg agggaacagg gagagagctc 180

gggcggccga gcgcgcagcc ctccgtgggc attgccgcct ggacctcccg gaaggcaccc 240

cgggccgcgg ccgccacccg tcccgccctc gttcggagct gagacgccgt cgccgcgcaa 300

gatgcccaag aagaagccga cgcccatcca gctgaacccg gcccccgacg gctcggcggt 360

gaacgggacc agctcggccg agaccaacct agaggctttg cagaagaagc tggaggagct 420

ggagctggat gagcagcagc ggaagcgcct cgaagctttc ctgacacaga agcagaaggt 480

gggcgagctg aaggacgatg actttgagaa gatcagtgag ctgggtgccg gcaatggcgg 540

tgtggtgttc aaggtctccc acaagccatc tggcctggtc atggcccgaa agcttatcca 600

cctggagatc aagccagcca tccgcaatca gatcatccgt gagctgcagg ttctgcacga 660

gtgcaactcg ccctacattg tgggcttcta tggggccttc tacagtgatg gcgagatcag 720

catctgcatg gagcacatgg atggaggttc cttggatcaa gtcctgaaga aagctggaag 780

aattcctgag caaattttag gaaaagttag cattgctgtg atcaaaggcc tgacatacct 840

gagggagaag cacaagatta tgcacagaga tgtcaagccc tccaacatcc tggtcaactc 900

ccgcggggag atcaagctct gtgactttgg ggtcagcggg cagctcatcg attccatggc 960

caactccttc gtgggcaccc ggtcctacat gtcgccagag agactgcagg gcacacacta 1020

ctcagtgcag tcggacatct ggagcatggg actgtcactg gtggagatgg cggttgggag 1080

gtaccccatc ccccctccag atgccaagga gctggagctg gtgttcgggt gccaggtgga 1140

aggagatgca gctgagatgc cgcccaggcc caggaccccc ggaagacccc tgagctcata 1200

tggaatggac agccggcctc ccatggcgat tttcgagctg ttggattaca tagtcaacga 1260

gccacctccc aaactgccca gtggagtctt cagtctggaa ttccaggact ttgtaaataa 1320

atgcttaata aagaaccctg cggagagagc agacttgaag cagctcatgg ttcatgcctt 1380

catcaagcgc tctgatgctg aggaggtgga cttcgcaggt tggctctgtg ccaccatcgg 1440

ccttaaccag cccagtaccc cgacccacgt ggccagcatctgagctgcgg cccggcccag 1500

acgtgctctg ccagcagccg ctatgctctg gcctctccct cgcttctctt cagacgtgcg 1560

tttcacctcc gaccagggtg cagacacagc atgtgccaag ctgtatttgt gttccttttc 1620

agtctttatt gccaccgtgt cacccgagtg gatttgcttt gtgcttaggg ctgtttgtgc 1680

tgatgatcac acacacgctg agctgaacag tgacacttgg tgatgtggtt gtcactgttc 1740

tcactccatg tggctggcct gttgcctcca gtgtctccag acttggggat gtctggtggc 1800

acttcccctg ccagggcatc tcctcagcag agagggaggc ctctgggccc ttgtccttgg 1860

cagtgcaagt ga 1872

<210>72

<211>393

<212>PRT

<213> Guinea pig

<400>72

Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp

1 5 10 15

Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala

20 25 30

Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys

35 40 45

Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly GluLeu Lys

50 55 60

Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly

65 70 75 80

Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Val Met Ala Arg

85 90 95

Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile

100 105 110

Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly

115 120 125

Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu

130 135 140

His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg

145 150 155 160

Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly

165 170 175

Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys

180 185 190

Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp

195 200 205

Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val

210 215 220

Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr

225 230 235 240

Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met

245 250 255

Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu

260 265 270

Leu Val Phe Gly Cys Gln Val Glu Gly Asp Ala Ala Glu Met Pro Pro

275 280 285

Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser

290 295 300

Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu

305 310 315 320

Pro Pro Pro Lys Leu Pro Ser Gly Val Phe Ser Leu Glu Phe Gln Asp

325 330335

Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu

340 345 350

Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu

355 360 365

Val Asp Phe Ala Gly Trp Leu Cys Ala Thr Ile Gly Leu Asn Gln Pro

370 375 380

Ser Thr Pro Thr His Val Ala Ser Ile

385 390

<210>73

<211>1257

<212>DNA

<213> dog

<400>73

ggagagcgag acacgggccg ctctccgctc ggagccggac gcgccttccc gcgtccaaaa 60

tgcccaagaa gaagccgacg cccatccagc tgaacccggc ccccgacggc tcggcggtga 120

acgggaccag ctcggcggag accaacctgg aggccttgca gaagaagctg gaggagctgg 180

agcttgatga gcagcagcgg aagcgccttg aggcctttct cacccagaag cagaaggtcg 240

gggaactgaa ggatgacgac ttcgagaaga tcagtgagct gggtgctggc aacggtggcg 300

tggtgttcaa ggtctcccac aagccgtccg gcctagtcat ggccagaaag ctaattcacc 360

tggagatcaa acctgcaatc cggaaccaga tcataaggga gctacaggtt ctacatgagt 420

gcaactcccc gtacatcgtg ggcttctatg gtgcattcta cagcgatggc gagatcagta 480

tctgcatgga gcacatggat gggggttcct tggatcaagt cctgaagaaa gctggaagaa 540

ttcctgaaca aattctagga aaagttagca tcgctgtaat aaaaggtctg acatacctga 600

gagagaagca caagattatg cacagagatg tcaagccttc caacatcctc gtgaactccc 660

gtggggagat caagctctgt gactttgggg tcagcgggca gctcattgac tccatggcca 720

actccttcgt gggcacaagg tcctacatgt cgccagaaag actccagggg actcattact 780

ccgtgcagtc ggacatctgg agcatggggc tctctctggt ggagatggca gttgggaggt 840

atcccatccc tcctccggat gccaaggagc tggagctgat gtttgggtgc caagtggagg 900

gagacgtggc tgagacccca cccagaccaa ggaccccggg aagacccctt agctcttatg 960

gaatggacag ccgaccgccc atggcaattt ttgagctgtt ggattacata gtcaacgagc 1020

cccctccaaa actgcccagt ggagtattca gtctggaatt tcaagatttt gtgaataaat 1080

gcttaataaa aaacccagca gagagagcag atctgaagca actcatggtt catgccttca 1140

tcaagagatc tgacggtgaa gaagtggatt ttgcaggttg gctctgctcc ccccattggc 1200

cttaaccagc ccagcacgcc gacccacgca gctggcgtct aactcgagtc tagagat 1257

<210>74

<211>381

<212>PRT

<213> dog

<400>74

Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp

1 5 10 15

Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala

20 25 30

Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys

35 40 45

Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys

50 55 60

Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly

65 70 75 80

Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Val Met Ala Arg

85 90 95

Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile

100 105 110

Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly

115 120 125

Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu

130 135 140

His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg

145 150 155 160

Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly

165 170 175

Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys

180 185 190

Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp

195 200 205

Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val

210 215 220

Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr

225 230 235 240

Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met

245 250 255

Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu

260 265 270

Leu Met Phe Gly Cys Gln Val Glu Gly Asp Val Ala Glu Thr Pro Pro

275 280 285

Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser

290 295 300

Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu

305 310 315 320

Pro Pro Pro Lys Leu Pro Ser Gly Val Phe Ser Leu Glu Phe Gln Asp

325 330 335

Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu

340 345 350

Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp Gly Glu Glu

355 360 365

Val Asp Phe Ala Gly Trp Leu Cys Ser Pro His Trp Pro

370 375 380

<210>75

<211>2095

<212>DNA

<213> Cat

<400>75

agccggcaag gagttgagcg tgcggggtgc ataggcgcgg gtcgtgggag atgaagctgg 60

agaggaccaa cctggaggcc ttgcagaaga agctggagga gctggagctc gatgagcagc 120

aacggaagcg cctggaggcc tttcttaccc agaagcagaa ggtcggggaa ttgaaggatg 180

acgacttcga gaagatcagc gagctgggcg ctggcaacgg tggtgtggtg ttcaaggtct 240

cccataagcc gtctggcctg gtcatggcca gaaagctaat tcacctggag atcaaacctg 300

caatccggaa ccagatcata agggagctgc aggttctaca tgagtgcaac tccccataca 360

tcgtgggctt ctatggcgcg ttctacagcg acggcgagat cagtatctgt atggagcaca 420

tggatggggg ttccttggat caagtcctga agaaagctgg aagaattcct gaacaaattt 480

taggaaaagt tagcattgct gtaataaaag gtctgacata cctgagggag aagcacaaga 540

ttatgcacag agatgtcaag ccttccaaca tcctagtgaa ctctcgtggg gagatcaagc 600

tctgtgactt tggggtcagc gggcagctca tcgactccat ggccaactcc ttcgtgggca 660

caaggtccta catgtcgcca gaaagactcc aggggactca ttactccgtg cagtcggaca 720

tctggagcat ggggctatct ctggttgaga tggcagtcgg gaggtatccc atccctcctc 780

ccgatgccaa ggagctggag ctgatgtttg ggtgccaagt ggagggagat gcggctgaga 840

cgccacccag gccgaggacc cccggaaggc ccctcagctc gtatggaatg gacagccgac 900

ctcccatggc aatttttgag ttgttggatt acatagtcaa cgagcctcct ccaaagctgc 960

ccagtggagt attcagtctg gaatttcaag attttgtgaa taaatgcctc ataaaaaacc 1020

cagcagagag agcagatctg aaacaactca tggttcatgc ctttatcaag agatctgatg 1080

gtgaggaagt ggattttgca ggttggctct gctccaccat cggccttaac cagcccagca 1140

caccgaccca cgcggccggc gtctaagtat ctgggaagca gcaaagagcg agtcccctgc 1200

ccagtggtgt gccattgtcg ctttcaggcc tctttgccat gcctgtctcc gttcagacgt 1260

gcatttcgcc tacgacaaag gatgaagaac acagcatgtg ccaaaattct atttgtgtct 1320

tttttaatat tactgtcatt tattctgtta tttccctaag tggattggct ttgtgcttgg 1380

ggctattttt gtgtatgttg atccaaacat gcgcaacgtt cagttacagt gaaaccttgg 1440

tgactgtggg tagtcattct tactgaaaat tgcactgctc ttcccccacc gtgactggct 1500

agctgcctgt agttttggga ttcttttgac acttggtggt actgcattct tgccgggcgc 1560

accttccttc tgttggggta ggagccttgt aagatccttc acaggcactg catgtgaagc 1620

atgctttgct gctatgaaaa agaacatcag aaagtataga tcttgttatt ttattatatt 1680

tttgcttttg gtgtagaatg aagcaatttc tgtcaaaatc tagccagagc ccttcactgc 1740

cacgatagct ggggcttcac cagtctgtct actgtgatga tttgtagact tctggttgta 1800

tttctatatt tattttaaaa tatattatgt gggatattta gtggtatgtg tctctttaag 1860

tttgaattag tgtttctaaa atgatggtta ctttgaatgt tacaaatgga tcaaggcatt 1920

aaaatgtatg agatttatct ttccccaaat ccaagtaccg atgctattgt aaacaacagt 1980

gtgtatagtg cctaagaatt gtatgaaaat ccttttaacc atttcaaccc agatgtttaa 2040

caaatctaat ctcttattct aataaatata ctatcaagtt aaaaggatga aaaaa 2095

<210>76

<211>371

<212>PRT

<213> Cat

<400>76

Met Lys Leu Glu Arg Thr Asn Leu Glu Ala Leu Gln Lys Lys Leu Glu

1 5 10 15

Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys Arg Leu Glu Ala Phe Leu

20 25 30

Thr Gln Lys Gln Lys Val Gly Glu Leu Lys Asp Asp Asp Phe Glu Lys

35 40 45

Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly Val Val Phe Lys Val Ser

50 55 60

His Lys Pro Ser Gly Leu Val Met Ala Arg Lys Leu Ile His Leu Glu

65 70 75 80

Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile Arg Glu Leu Gln Val Leu

85 90 95

His Glu Cys Asn Ser Pro Tyr Ile Val Gly Phe Tyr Gly Ala Phe Tyr

100 105 110

Ser Asp Gly Glu Ile Ser Ile Cys Met Glu His Met Asp Gly Gly Ser

115 120 125

Leu Asp Gln Val Leu Lys Lys Ala Gly Arg Ile Pro Glu Gln Ile Leu

130 135 140

Gly Lys Val Ser Ile Ala Val Ile Lys Gly Leu Thr Tyr Leu Arg Glu

145 150 155 160

Lys His Lys Ile Met His Arg Asp Val Lys Pro Ser Asn Ile Leu Val

165 170 175

Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp Phe Gly Val Ser Gly Gln

180 185 190

Leu Ile Asp Ser Met Ala Asn Ser Phe Val Gly Thr Arg Ser Tyr Met

195 200 205

Ser Pro Glu Arg Leu Gln Gly Thr His Tyr Ser Val Gln Ser Asp Ile

210 215 220

Trp Ser Met Gly Leu Ser Leu Val Glu Met Ala Val Gly Arg Tyr Pro

225 230 235 240

Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu Leu Met Phe Gly Cys Gln

245 250 255

Val Glu Gly Asp Ala Ala Glu Thr Pro Pro Arg Pro Arg Thr Pro Gly

260 265 270

Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser Arg Pro Pro Met Ala Ile

275 280 285

Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu Pro Pro Pro Lys Leu Pro

290 295 300

Ser Gly Val Phe Ser Leu Glu Phe Gln Asp Phe Val Asn Lys Cys Leu

305 310 315 320

Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu Lys Gln Leu Met Val His

325 330 335

Ala Phe Ile Lys Arg Ser Asp Gly Glu Glu Val Asp Phe Ala Gly Trp

340 345 350

Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro Ser Thr Pro Thr His Ala

355 360 365

Ala Gly Val

370

<210>77

<211>2405

<212>DNA

<213> cattle

<400>77

ccggttgact gagggagagt gggagggaat cccgggctgc cgagctgcgc cggcggggaa 60

gcccttcggt tccctgtgca ctgagcaagt gggccggggg gttcccccag accgccactg 120

gccctttggc cctgacggga ccgcgcaggg cgcgcccccc gaaggcagcc ttcgggcttg 180

cggcccagac ttggccccgc gaggccggcg cggggcagct cagagggagg aagctagagg 240

ggccgccctc agagttggga gcgcctttcc tgggtccaaa atgcccaaga agaagccgac 300

gcccatccag ctgaacccgg ccccggacgg ctccgcggtt aacgggacca gctcggcgga 360

gaccaacctg gaggccttgc agaagaagct ggaggagctg gagctcgatg aacagcagcg 420

gaagcgcctc gaggcctttc tgacccagaa gcagaaggtg ggggaactga aggatgatga 480

ctttgagaag atcagtgagc tgggtgccgg caatggaggt gtggtgttca aggtctccca 540

caagccgtcc ggacttgtta tggccagaaa gctaattcac ctggagatca aacctgccat 600

ccggaaccag atcataaggg agctgcaggt tctccatgag tgcaactcgc cttatatcgt 660

gggcttctac ggggcgttct acagcgacgg cgagatcagc atctgcatgg agcacatgga 720

tgggggttcc ttggatcaag ttctgaagaa agctggaaga attcctgaac aaattttagg 780

aaaagttagc attgctgtaa taaaaggcct gacatacctg agggagaagc acaagattat 840

gcacagagat gtcaagccgt ccaacatcct agtgaacagc cgtggagaga tcaagctctg 900

tgactttggg gtcagcgggc agctcatcga ctccatggcc aactccttcg tgggcaccag 960

gtcctacatg tcgccagagc gactccaggg gacccattac tccgtgcagt cggacatctg 1020

gagcatgggg ctctctctgg ttgagatggc tgtcgggagg tatcccatcc ctcctccaga 1080

tgccaaggag ctggagctga tgtttgggtg ccaggtggag ggagatgcgg ctgagacccc 1140

gcccaggcca aggacccccg ggaggcccct cagctcttat ggaatggaca gccgacctcc 1200

aatggcaatt tttgagttgt tggattacat agtcaatgag cctcctccaa aactgcccag 1260

tggagtattc agtctggaat ttcaagattt tgtgaataaa tgcttaataa aaaaccccgc 1320

agagagagca gatttgaagc aactcatggt tcatgctttt atcaagagat ctgatgctga 1380

ggaagtggat tttgcaggtt ggctctgctc caccatcggc cttaaccaac ccagcacacc 1440

cacccatgcg gctggcgtct aagtggttgg gaagcagcag tccctgccca agggcatgca 1500

ctgttgcttc cgggcagcct tcccatgcct gtctctgttc agacgtgcat ttcacctatg 1560

acaaaggatg aagaacacag catgtgccaa aattctattt gtgtcatttt caatattatc 1620

atctttactc ttattactat tgttattccc ctaagtggat tggctttgtg cttggggcta 1680

tttttgtgta tattgatgat gaagacatgt gcaatgtaga attacagtga aactctggtg 1740

actgtgggta gtcattctta ctgaaaactg cactgctttc ccacaccatg aactggctgg 1800

tcgcctctat tttcgggatt ctttgacact tggtggtact tcattcttgc caggcatacc 1860

ttctaactga gtaggaagga gccttgtaag atccttcaca ggcagtgcat gtgaagcatg 1920

ctttgctgct ataaaaatga gcatcagaaa gtgtgtatca tgttatttta ttatgttctt 1980

gcttttggtg tagaattcag caaattttca tcaaaatcta gccagagccc ttcactgcca 2040

tgatagctgg ggcttcacca gtctgtctac tgtgatgatt tgtagacttc tggttgtatt 2100

tctgtattta tttttaaatc taccgtgtgg atatttagtg ctatgtctct ttaagtttgg 2160

attagtgttt ctaaaatggt ggagttgctc tgaatgttac aaatggatca aggcattaaa 2220

atgaatgaga tctacctttc accaagtact gatgctattg taaacaacag tgtgtatagt 2280

gcctaacaac tgtatgaaaa tccttttacc attttaatcc agatgtttaa caagcctaat 2340

ctcttactct aataaatata ctatcaaatt caaaggaaaa aaaaaaaaaa aaaaaaaaaa 2400

aaaaa 2405

<210>78

<211>393

<212>PRT

<213> cattle

<400>78

Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro Ala Pro Asp

1 5 10 15

Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala

20 25 30

Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys

35 40 45

Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys

50 55 60

Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly

65 70 75 80

Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Val Met Ala Arg

85 90 95

Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile

100 105 110

Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly

115 120 125

Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu

130 135 140

His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg

145 150 155 160

Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly

165 170 175

Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys

180 185 190

Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp

195 200 205

Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val

210 215 220

Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr

225 230 235 240

Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met

245 250 255

Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu

260 265 270

Leu Met Phe Gly Cys Gln Val Glu Gly Asp Ala Ala Glu Thr Pro Pro

275 280 285

Arg Pro Arg Thr Pro Gly Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser

290 295 300

Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu

305 310 315 320

Pro Pro Pro Lys Leu Pro Ser Gly Val Phe Ser Leu Glu Phe Gln Asp

325 330 335

Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu

340 345 350

Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu

355 360 365

Val Asp Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro

370 375 380

Ser Thr Pro Thr His Ala Ala Gly Val

385 390

<210>79

<211>2146

<212>DNA

<213> horse (Equus caballus)

<400>79

caagtgggaa agcttgggat gcgtggagga gccagctact agtttagtgt gctgggtgcg 60

tcggtgcagg tcgcaggaga tgaagccgga gaggaccaac ctggaggcct tgcagaagaa 120

gctggaggag ctggagctcg atgaacagca gcgaaagcgc cttgaggcct ttcttactca 180

gaagcagaag gttggggaac tgaaggatga tgactttgag aagatcagtg agctgggtgc 240

tggcaatggt ggtgtggtat tcaaggttgc ccacaaaccg tctggtttgg tcatggccag 300

aaagctaatt cacctggaga tcaagcctgc aatccggaac cagatcataa gggagctgca 360

ggttctacat gagtgcaact ccccatatat tgtgggcttc tatggcgcat tctacagcga 420

tggtgagatc agcatctgca tggagcacat ggatgggggt tccttggatc aagtcctaaa 480

gaaagctgga agaattcctg agcaaatttt aggaaaagtt agcattgctg taataaaagg 540

cctgacgtat ctgagggaga agcacaagat tatgcacaga gatgtcaagc cctccaacat 600

cctagtgaac tcccgtgggg agatcaagct gtgtgatttt ggggtcagcg ggcagctcat 660

cgactccatg gccaactcct tcgtgggcac aaggtcttac atgtcgccgg aaagactcca 720

ggggactcat tattcagtgc agtcggacat ctggagcatg gggctctctc tggttgagat 780

ggcggtcggg aggtatccca tccctcctcc agatgccaag gagctggagc tgatgtttgg 840

gtgccaagtg gagggagatg cggctgagac tccgcctagg ccaaggaccc ctggaagacc 900

cctcagctct tatggaatgg acagccgacc tcctatggca atttttgagt tactggatta 960

catagtcaac gagcctcctc ccaagctgcc cagtggagta ttcagtctgg aatttcagga 1020

ttttgtgaat aaatgcttaa tcaaaaaccc tgcagagaga gcagatttga agcaactcat 1080

ggttcacgct tttatcaaga gatctgatgc cgaggaagtg gattttgcag gttggctctg 1140

ctccaccatt ggccttaacc agcccagcac accaacccac gcggctggcg tctaagcgtt 1200

tgggaagcag caaaaagcga gccccctgcc gcgtggtgtg ccatgttgct ttcgggcctc 1260

cttcccatgc ctgtctgttc acacgtgcat ttcacctgtg acaaaggatg aagaacacag 1320

catgtgccaa aattctattt gtgtcatttt taatagtact gtctttattc ttattactat 1380

tgttattccc ctaagtggat tggctttgtg cttgggacta ttttgtgtat gttgatgatc 1440

aaaacatgcg caatgttgaa ttaccgtgaa actggtgact gtgggtagtc cttcttattg 1500

aaaattgcac tgctcttccc tccctgtcac tggctggctg cctgtatttc tggggttctt 1560

tgacacttgg tggtacttca ttcttgcagg gcatacctcc tattcgagta ggaaggagcc 1620

tttaagatcc ttcacaggca gtgcatgtga agcatgcttt gctgctatga aaatgagcat 1680

cagaaagtgt atatcatgtt attttattat tattatgttt ttgcttttgg tgtagaattc 1740

agcaatttcc atcaagatct agccagagcc cttcactgcc atgatagctg gggcttcacc 1800

agtctgccta ctgtgatgat ttgtagactt ctggttgtat ttctatattt atttttaaat 1860

atactgtgtg ggatatttag tggtatatgt ctctctaagt ttggagtggt gtttctaaaa 1920

tggagttact ttgaatgtta tagatggatc aaggcataaa atgtatgaga tttatttttc 1980

cccaaatcca aatactgatg ctattgtaaa caacaaacag tgtgtatagt gcctaaaaat 2040

tgtatgaaag tccttttaac cattttaatc cagatgttta acaaatctaa tctcttattc 2100

taataaatat actatcaagt taaacggaca aaagatttct actttc 2146

<210>80

<211>371

<212>PRT

<213> horse

<400>80

Met Lys Pro Glu Arg Thr Asn Leu Glu Ala Leu Gln Lys Lys Leu Glu

1 5 10 15

Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys Arg Leu Glu Ala Phe Leu

20 25 30

Thr Gln Lys Gln Lys Val Gly Glu Leu Lys Asp Asp Asp Phe Glu Lys

35 40 45

Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly Val Val Phe Lys Val Ala

50 55 60

His Lys Pro Ser Gly Leu Val Met Ala Arg Lys Leu Ile His Leu Glu

65 70 75 80

Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile Arg Glu Leu Gln Val Leu

85 90 95

His Glu Cys Asn Ser Pro Tyr Ile Val Gly Phe Tyr Gly Ala Phe Tyr

100 105 110

Ser Asp Gly Glu Ile Ser Ile Cys Met Glu His Met Asp Gly Gly Ser

115 120 125

Leu Asp Gln Val Leu Lys Lys Ala Gly Arg Ile Pro Glu Gln Ile Leu

130 135 140

Gly Lys Val Ser Ile Ala Val Ile Lys Gly Leu Thr Tyr Leu Arg Glu

145 150 155 160

Lys His Lys Ile Met His Arg Asp Val Lys Pro Ser Asn Ile Leu Val

165 170 175

Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp Phe Gly Val Ser Gly Gln

180 185 190

Leu Ile Asp Ser Met Ala Asn Ser Phe Val Gly Thr Arg Ser Tyr Met

195 200 205

Ser Pro Glu Arg Leu Gln Gly Thr His Tyr Ser Val Gln Ser Asp Ile

210 215 220

Trp Ser Met Gly Leu Ser Leu Val Glu Met Ala Val Gly Arg Tyr Pro

225 230 235 240

Ile Pro Pro Pro Asp Ala Lys Glu Leu Glu Leu Met Phe Gly Cys Gln

245 250 255

Val Glu Gly Asp Ala Ala Glu Thr Pro Pro Arg Pro Arg Thr Pro Gly

260 265 270

Arg Pro Leu Ser Ser Tyr Gly Met Asp Ser Arg Pro Pro Met Ala Ile

275 280 285

Phe Glu Leu Leu Asp Tyr Ile Val Asn Glu Pro Pro Pro Lys Leu Pro

290 295 300

Ser Gly Val Phe Ser Leu Glu Phe Gln Asp Phe Val Asn Lys Cys Leu

305 310 315 320

Ile Lys Asn Pro Ala Glu Arg Ala Asp Leu Lys Gln Leu Met Val His

325 330 335

Ala Phe Ile Lys Arg Ser Asp Ala Glu Glu Val Asp Phe Ala Gly Trp

340 345 350

Leu Cys Ser Thr Ile Gly Leu Asn Gln Pro Ser Thr Pro Thr His Ala

355 360 365

Ala Gly Val

370

<210>81

<211>2274

<212>DNA

<213> Chicken

<400>81

ctaaccaggc gggagctgtc ggtgcggagc tcggtgtcgc tccgccgggc aggccgggtc 60

gacggccgcg ctgtgccgga gcggcagcgt cgcgggctcg gctccttctc ggggaggcgg 120

ccgcgcgctg ctccggcgct gaggggcggc cccgaagttt gcttcgcgtc gggaagtccg 180

tcggacctgg ccgaagtggg gccgcggccg ctccgtccgt cacgctctgc gctggccggg 240

gggcaacatg cccaagaaga agccagggcc gatccagctc aaccccgctc cggatggctc 300

cgccgtcaac gggaccagct ctgccgagac aaacttggaa gctcttcaga agaagctgga 360

agagctagag ctggatgaac agcaaaggaa gcgccttgaa gctttcctta cccagaaaca 420

aaaagttggg gagctgaagg atgatgactt cgagaagatc agtgagctgg gagcagggaa 480

tggcggggtg gtgttcaaag tatctcacaa gccttctggc ctcatcatgg caagaaaatt 540

gattcatcta gaaatcaagc cagctattcg aaaccagatc atccgtgagc tgcaggttct 600

acatgagtgc aattcaccat acatagtggg cttttatgga gctttttaca gtgatgggga 660

aatcagcatt tgcatggaac acatggatgg tggctcattg gatcaagtgc tgaaaaaggc 720

tggaagaatt ccagagcaga tactgggcaa agtcagcatt gcagtaataa aaggactcac 780

atatctgaga gaaaagcata aaataatgca cagagatgtt aaaccatcta acatcttggt 840

aaactctaga ggtgagatca agctttgtga ttttggtgtc agtggacaac tgatagattc 900

tatggcaaac tcatttgttg gcacgcgctc ctacatgtct ccggaaagac tgcagggaac 960

tcattattca gtgcagtcag atatatggag tatggggctg tctctggtag aaatggccat 1020

tggcagatac ccgattcctc ctcctgactc taaggagctc gagttgatgt ttggctgccc 1080

ggtagaggga gattctccag tcacagagac ctcacccagg caaagaacac ctggtcgacc 1140

aatgagctcc tatggaccag acagcagacc cccgatggca atctttgaac ttctggatta 1200

catcgtcaat gagccacctc caaaactgcc caatggtgtc tttggttctg aatttcaaga 1260

ttttgttaac aaatgtttaa ttaaaaatcc tgctgagaga gctgatttga agcagctgat 1320

gattcacgct ttcattaaga gatctgaagc agaggaggtg gattttgcag gatggctttg 1380

ctcaaccata ggccttaacc aaccgagtac acccacgcat gctgctggag tctgaatgtg 1440

gaagagcaaa tcctgtcccg tacatctgtt aacagcgcta ctttggtcct atttcctaag 1500

cttgtacctg ttcaaacatg tatttcacct cttaaggaag aatgtcttta tagcatgtgc 1560

caaattgttt tcaattttgt catcaactaa ttggtattgt actgggttac atttgtttgc 1620

tgaccaaaat gtaaaatgtt taagttacag tgcttgctga ttttaagtga ttatggaatt 1680

atggatattc tttcttaatg aaaatatcac tgggggggaatttacccctg gattgtttga 1740

actttatcaa gactctttgt aaactgttgg tacttcagtc atgcttacct aatctcccat 1800

gcaaaaaaag gggtagggat gctccaaaac tgtatctgtt gagcatgctt ttgctgctgc 1860

caaactgtat cttggaagtt aggcctaatg gttccaattt ggtgttgtgt agagatcact 1920

ctttccaggt aaagaaggta agagctctgc attccttggg atggacaggg cagtatccta 1980

cttgtagact tgttcatatt tctatattta tttttaaaat gtatcatcat acttggattt 2040

agtgatatat gtctttccaa ttgattttta aaggttagct ctcagaagcg tcctacagaa 2100

tcatgacaaa gatctgggct ttcttttaac cttaagattc atgacagctg tgtttggtgt 2160

ctaaaatgta tgaagatcct ctattgtttt attctctcag atgtttagca atggtttctc 2220

ttaataaata tattatcaag taaaaaaaaa aaaaaaataa aaaaaaaaaa aaaa 2274

<210>82

<211>395

<212>PRT

<213> Chicken

<400>82

Met Pro Lys Lys Lys Pro Gly Pro Ile Gln Leu Asn Pro Ala Pro Asp

1 5 10 15

Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn Leu Glu Ala

20 25 30

Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp Glu Gln Gln Arg Lys

35 40 45

Arg Leu Glu Ala Phe Leu Thr Gln Lys Gln Lys Val Gly Glu Leu Lys

50 55 60

Asp Asp Asp Phe Glu Lys Ile Ser Glu Leu Gly Ala Gly Asn Gly Gly

65 70 75 80

Val Val Phe Lys Val Ser His Lys Pro Ser Gly Leu Ile Met Ala Arg

85 90 95

Lys Leu Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn Gln Ile Ile

100 105 110

Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr Ile Val Gly

115 120 125

Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile Cys Met Glu

130 135 140

His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys Ala Gly Arg

145 150 155 160

Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile Ala Val Ile Lys Gly

165 170175

Leu Thr Tyr Leu Arg Glu Lys His Lys Ile Met His Arg Asp Val Lys

180 185 190

Pro Ser Asn Ile Leu Val Asn Ser Arg Gly Glu Ile Lys Leu Cys Asp

195 200 205

Phe Gly Val Ser Gly Gln Leu Ile Asp Ser Met Ala Asn Ser Phe Val

210 215 220

Gly Thr Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly Thr His Tyr

225 230 235 240

Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu Val Glu Met

245 250 255

Ala Ile Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ser Lys Glu Leu Glu

260 265 270

Leu Met Phe Gly Cys Pro Val Glu Gly Asp Ser Pro Val Thr Glu Thr

275 280 285

Ser Pro Arg Gln Arg Thr Pro Gly Arg Pro Met Ser Ser Tyr Gly Pro

290 295 300

Asp Ser Arg Pro Pro Met Ala Ile Phe Glu Leu Leu Asp Tyr Ile Val

305 310 315 320

Asn Glu Pro Pro Pro Lys Leu Pro Asn Gly Val Phe Gly Ser Glu Phe

325 330 335

Gln Asp Phe Val Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg Ala

340 345 350

Asp Leu Lys Gln Leu Met Ile His Ala Phe Ile Lys Arg Ser Glu Ala

355 360 365

Glu Glu Val Asp Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu Asn

370 375 380

Gln Pro Ser Thr Pro Thr His Ala Ala Gly Val

385 390 395

Claims

1. A method of treating a subject with unresectable or metastatic BRAF600 mutation-positive melanoma comprising administering to the subject 600mg BID of BVD-523 or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the mutation is BRAF^V600EAnd (4) mutation.

3. The method of claim 1, wherein the mammal is selected from the group consisting of a human, a primate, a farm animal, and a domestic animal.

4. The method of claim 1, wherein the mammal is a human.

5. The method of claim 1, wherein the melanoma has MAPK activity.

6. A composition for use in treating a subject with unresectable or metastatic BRAF600 mutation-positive melanoma, the composition comprising 600mg BVD-523 or a pharmaceutically acceptable salt thereof and optionally a pharmaceutically acceptable carrier, adjuvant or vehicle.

7. The composition of claim 6, wherein the subject is a mammal.

8. The composition according to claim 6, wherein the mammal is selected from the group consisting of humans, primates, farm animals and domestic animals.

9. The composition according to claim 6, wherein the mammal is a human.

10. The composition of claim 6, wherein the melanoma has MAPK activity.

11. The composition of claim 6, wherein the mutation is BRAF^V600EAnd (4) mutation.