WO2003000893A2 - Nucleic acids encoding g protein-coupled receptors - Google Patents

Abstract

Nucleic acids encoding G protein-coupled receptors are disclosed, and methods of using same.

Description

NUCLEIC ACIDS ENCODING G PROTEIN-COUPLED RECEPTORS

RELATED APPLICATIONS

This application claims the benefit of and priority to U.S Provisional Application 60/301,095, filed June 26, 2001 and to U.S. Provisional Application 60/333,185, filed November 6, 2001, the entire teachings of which are incorporated herein by reference.

BACKGROUND OF THE INVENTION

G protein-coupled receptors ("GPCRs") are a superfamily of intrinsic transmembrane cell-surface receptors that mediate the transmission of extracellular signals into the cell to produce a cellular response. There are thought to be anywhere from 400 to over 1000 different members of this family. GPCRs are intrinsic membrane proteins, and operate by a common transduction mechanism. In their inactive state, the GPCRs bind to the G proteins. Upon activation, they stimulate guanine nucleotide exchange on the G proteins, resulting in the release of GDP and the binding of GTP. The G-protein then dissociates from the GPCR, and interacts with the adenylate cyclases, which catalyze the conversion of ATP into cAMP. The cAMP then acts as a second messenger. The G proteins can cause intracellular coupling of the GPCRs with various intracellular enzymes, ion channels and transporters.

GPCRs (and perforce, G proteins) are involved in an enormous range of biological processes, and have been found to regulate such processes as hydrolysis

+ 2+ of plasma membrane phospholipids, the K and Ca ion channels, yeast mating signals, the signaling by cholera and pertussis toxins, and proliferation in some cancers (e.g., pituitary, adrenal, ovarian). The signal can be endogenous or exogenous or, in the case of rhodopsin receptors, the stimulus can be light. Many drugs bind to a GPCR and either produce a response or block the actions of the normal signal. The GPCR superfamily includes the cannabinoid and opioid receptors, chemokine, histamine, angiotensin, neurotensin, vasopressin, calcitonin, dopamine, glutamate and bombesin receptors, taste and odorant receptors, and many others.

SUMMARY OF THE INVENTION

The present invention relates to human G protein-coupled receptor (GPCR) genes, particularly nucleic acids comprising GPCR genes, and the amino acids encoded by such nucleic acids. These sequences are shown in Tables I and II. In Tables I and π, each GPCR entry lists the name (e.g., "MOOSE00162"), the University of California at Santa Cruz contig designation from which the sequence was analyzed (e.g., "ctgl4797"), the exon locations (e.g., "448003 . . 448092, . . ."), followed by the amino acid sequence and the nucleic acid sequence.

Sub-family information on the sequences is shown in Table El. For each sequence, the following information is provided: the University of California at Santa Cruz contig designation from which the sequence was analyzed (e.g., "ctgl4797"), the name (e.g., "MOOSE00162"), and the subfamily to which the sequence appears to belong. The assignments were made on the basis of the best E- value with which the sequence aligned. Sequences listed as "Class A Orphan" are those that have been characterized, and are known to bind GPCR-proteins.

In one embodiment, the isolated nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-124 (odd numbers), as shown in Tables I and π, and the complements thereof. The invention further relates to a nucleic acid molecule which hybridizes under high stringency conditions to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-124 (odd numbers), as shown in Tables I and II, and the complements thereof. The invention additionally relates to isolated nucleic acid molecules (e.g., cDNA molecules) encoding a GPCR polypeptide (e.g., encoding a polypeptide selected from the group consisting of SEQ ID NOs: 1-124 (even numbers), as shown in Tables I and II).

The invention further provides a method for assaying a sample for the presence of a nucleic acid molecule comprising all or a portion of a GPCR in a sample, comprising contacting said sample with a second nucleic acid molecule comprising a nucleotide sequence encoding a GPCR polypeptide (e.g., one of SEQ ID NOs: 1-124 (odd numbers), as shown in Tables I and II, or the complement of one of SEQ ID NOs: 1-124 (odd numbers); a nucleotide sequence encoding one of SEQ ID NOs: 1-124 (even numbers), as shown in Tables I and TT), or a fragment or derivative thereof, under conditions appropriate for selective hybridization. The invention additionally provides a method for assaying a sample for the level of expression of a GPCR polypeptide, or fragment or derivative thereof, comprising detecting (directly or indirectly) the level of expression of the GPCR polypeptide, fragment or derivative thereof. The invention also relates to a vector comprising an isolated nucleic acid molecule of the invention operatively linked to a regulatory sequence, as well as to a recombinant host cell comprising the vector. The invention also provides a method for preparing a polypeptide encoded by an isolated nucleic acid molecule described herein (a GPCR polypeptide), comprising culturing a recombinant host cell of the invention under conditions suitable for expression of said nucleic acid molecule. The invention further provides an isolated polypeptide encoded by isolated nucleic acid molecules of the invention (e.g., GPCR polypeptide), as well as fragments or derivatives thereof. In a particular embodiment, the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-124 (even numbers), as shown in Tables I and II. The invention also relates to an isolated polypeptide comprising an amino acid sequence which is greater than about 90 percent identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-124 (even numbers), preferably about 95, 96, 97, 98 and 99 percent identical.

The invention also relates to an antibody, or an antigen-binding fragment thereof, which selectively binds to a polypeptide of the invention, as well as to a method for assaying the presence of a polypeptide encoded by an isolated nucleic acid molecule of the invention in a sample, comprising contacting said sample with an antibody which specifically binds to the encoded polypeptide.

The invention further relates to methods of diagnosing a predisposition to a condition mediated by GPCRs. The methods of diagnosing such a predisposition in an individual include detecting the presence of a mutation in GPCR, as well as detecting alterations in expression of a GPCR polypeptide, such as the presence of different splicing variants of GPCR polypeptides. The alterations in expression can be quantitative, qualitative, or both quantitative and qualitative.

The invention additionally relates to an assay for identifying agents that alter (e.g., enhance or inhibit) the activity or expression of one or more GPCR polypeptides. For example, a cell, cellular fraction, or solution containing a GPCR polypeptide or a fragment or derivative thereof, can be contacted with an agent to be tested, and the level of GPCR polypeptide expression or activity can be assessed. The activity or expression of more than one GPCR polypeptides can be assessed concurrently (e.g., the cell, cellular fraction, or solution can contain more than one type of GPCR polypeptide, such as different splicing variants, and the levels of the different polypeptides or splicing variants can be assessed).

In another embodiment, the invention relates to assays to identify polypeptides that interact with one or more GPCR polypeptides. In a yeast two-hybrid system, for example, a first vector is used which includes a nucleic acid encoding a DNA binding domain and also an GPCR polypeptide, splicing variant, or fragment or derivative thereof, and a second vector is used which includes a nucleic acid encoding a transcription activation domain and also a nucleic acid encoding a polypeptide which potentially may interact with the GPCR polypeptide, splicing variant, or fragment or derivative thereof (e.g., a GPCR polypeptide binding agent or receptor). Incubation of yeast containing both the first vector and the second vector under appropriate conditions allows identification of polypeptides which interact with the GPCR polypeptide or fragment or derivative thereof, and thus can be agents which alter the activity of expression of an GPCR polypeptide.

Agents that enhance or inhibit GPCR polypeptide expression or activity are also included in the current invention, as are methods of altering (enhancing or inhibiting) GPCR polypeptide expression or activity by contacting a cell containing GPCR and/or polypeptide, or by contacting the GPCR polypeptide, with an agent that enhances or inhibits expression or activity of GPCR or polypeptide.

Additionally, the invention pertains to pharmaceutical compositions comprising the nucleic acids of the invention, the polypeptides of the invention, and/or the agents that alter activity of GPCR polypeptide. The invention further pertains to methods of treating conditions mediated by GPCRs, by administering GPCR therapeutic agents, such as nucleic acids of the invention, polypeptides of the invention, the agents that alter activity of GPCR polypeptide, or compositions comprising the nucleic acids, polypeptides, and/or the agents that alter activity of GPCR polypeptide.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to nucleic acids comprising G protein-coupled receptors ("GPCRs"), and the GPCR amino acids encoded by those nucleic acids. The GPCR superfamily includes the receptors for many important signaling pathways, including, but not limited to, hormone receptors, growth factors, viral receptors, neuroreceptors, etc., such as acetylcholine, adrenocorticotropin (ACTH), adenosine, α-adrenergic receptors, β-adrenergic receptors, angiotensin, bombesin, bradykinin, C5a, calcitonin, cAMP, cannabinoid, C-C chemokine, cholecystokinin/gastrin (CCK/gastrin), cytomegalovirus, dopamine, endothelial differentiation gene-1, endothelin, formyl peptide, glutamate (metabotropic), gonadotropin-releasing hormone, growth hormone-releasing hormone, histamine, 5- hydroxytryptamine, interleukin-8, kinin, luteinizing hormone/follicle-stimulating hormone/thyroid-stimulating hormone (LH/FSH/TSH), mas, melanocortin, muscarinic, neuropeptide Y, neurotensin, odorant, opioid, opsins, parathyroid hormone, platelet-activating factor (PAF), prolactin, prostaglandin E, rhodopsins, secretin, serotonin, somatostatin, tachykinin, taste, testis specific, thrombin, thromboxine A₂, thyrotropin-releasing hormone (TRH), tyramine/octopamine, vasoactive intestinal peptide (VIP), vasopressin, viral and yeast mating factor. These receptors are involved in the treatment of infections and various diseases and conditions, including, but not limited to, bacterial, fungal, protozoan and viral infections, particularly infections caused by HTV-1 or HJV-2; cancers; diabetes; asthma; Parkinson's disease; both acute and congestive heart failure; hypotension; hypertension; urinary retention; osteoporosis; angina pectoris; myocardial infarction; ulcers; asthma; allergies; benign prostatic hypertrophy (benign prostatic hyperplasia); chronic renal failure; renal disease; impaired glucose tolerance; seizure disorder; depression; anxiety; obsessive compulsive disorder; affective neurosis/disorder; depressive neurosis/disorder; anxiety neurosis; dysthymic disorder; behavior disorder; mood disorder; shizophrenia; psychosexual dysfunction; sex disorder; sexual disorder; disturbed biological and circadian rhythms; feeding disorders, such as anorexia, bulimia, cachexia, and obesity; Cushing's syndrome/disease; basophil adenoma; prolactinoma; hyperprolactinemia; hypopituitarism; hypophysis tumor/adenoma; hypothalamic diseases; Froehlich's syndrome; adenohypophysis disease; hypophysis disease; hypophysis tumor/adenoma; pituitary growth hormone; adenohypophysis hypofunction; adrenohpophysis hyperfuunction; hypothalamic hypogonadism; Kallman's syndrome (anosmia, hyposmia); functional or psychogenic amenorrhea; hypopituitarism; hypothalamic hypothyroidism; hypothalamic-adrenal dysfunctions; idiopathic hyperprolactinemia; hypothalamic disorders of growth hormone deficiency; idiopathic growth hormone deficiency; dwarfϊsm; gigantism; acromegaly; disturbed biological and circadian rhythms; and sleep disturbances associated with such diseases as neurological disorders, heart and lung diseases, mental illness, and addictions; migraine; hyperalgesia; enhanced or exaggerated sensitivity to pain, such as hyperlgesia, causalgia and allodynia; acute pain; burn pain; atypical facial pain; neuropathic pain; back pain; complex regional pain syndromes I and U; arthritic pain; sports injury pain; pain related to infection, e.g., HTV, post-polio syndrome, and post-herpetic neuralgia; phantom limb pain; labour pain; cancer pain; post-chemotherapy pain; post-stroke pain; post-operative pain; neuralgia; and tolerance to narcotics or withdrawal from narcotics; sleep disorders; sleep apnea; narcolepsy; insomnia; parasomnia; jet-lag syndrome; and other neurodegenerative disorders, which includes nosological entities such as disinhibition-dementia-parkinsonism-amyotrophy complex; pallido-ponto-nigral degeneration; and dyskinesias, such as Huntington's disease or Gilles dela Tourett's syndrome.

With the availability of complete genomic sequences for many organisms today, including Homo sapiens, it has become clear that there is a need for data mining techniques to extract the information in them, e.g., gene prediction programs. Of these, the most successful ones are those based on the comparison of known protein or protein-derived information, or those that use expressed sequence tags (ESTs) to predict gene location and structure.

One such algorithm is Gene Wise. It bases its exon prediction on the use of Hidden Markov Models (HMMs) of proteins to be compared against a genomic sequence, so that the translation of the sequence will match the model in a similar way to other HMM profile searches (Eddy, Curr. Opin. Struct. Biol. 6(3):361-5, 1996), and allowing the presence of long insertions as long as they include donor and acceptor site sequences at both ends. To take advantage of the algorithm, the models for different protein families must be built so that they represent the full-length sequences instead of the most common features in them. This is a major difference with existing HMM databases such as Pfam (Sonnhammer et al, Proteins 28(3):405-20, 1997), in which each model is built to represent a family of proteins as broad as possible with minimum overlap between them.

In the present approach, the sequences were subdivided in several families so that the similarity inside of a group of them was over 50%. Given this approach, there are several points of overlap between different families when analyzing a sequence, so the discrimination must be done after the search is completed. Several resources that include expert-supervised classifications are used to select the best groups of sequences, e.g., the GPCR database (Horn et al, Nucleic Acids Res. 26(l):275-9, 1998), PKR (Smith et al, Trends Biochem. Sci. 22(1 1):444- 6, 1997), NuclearRdb (Horn et al, Nucleic Acids Res. 29:346-349, 2001), IOCH (Le Novere et al, Nucleic Acids Res. 27(l):340-2, 1999), Enzyme (Bairoch, Nucleic Acids Res. 28:304-305, 2000) and Swiss-Prot (Bairoch et al. , Nucleic Acids Res.

28:45-48, 2000). When none is available, or the sequences included in some groups are too disrinatly related, the grouping must be done manually, using the ClustalW (Thompson et al, Nucleic Acids Res. 22:4673-4680, 1994) package to measure the distance between different sequences. The present model was built from multiple sequence alignments of the different protein families obtained with DiAlign 2 (Morgenstern, Bioinformatics 15(3):211-8, 1999). DiAlign works based on segment-to-segment comparisons instead of arbitrary thresholds for gap opening and extension, which makes it ideally suited for building models that represent an entire, full-length sequence, since the alignments built this way have more match states that would be assigned as insertion states when using other alignment algorithms. The models were built using the standard HMMer package. To search for new genes, a genome-wide scan was done on the University of California at Santa Cruz sequences, using the Gene Wise algorithm. It translates the genomic sequence on the fly to proteins and can therefore maintain a reading frame through insertions and deletions. The algorithm also rewards gaps in the genomic sequence relative to the model if they are encapsulated within introns, like splice structure.

For each superfamily of proteins, a classification was obtained in which the sequences are grouped by length and similarity. Each one of these groups was then used to build a HMM profile representing this group of sequences. This approach aims to have models that can represent the full length of the encoded proteins for a whole range of proteins, without being too specific for any one of them or being too general, as would be a HMM built for large groups of sequences. This classification was based either on existing expert-supervised classifications, or by retrieval of sequences and classification based on pairwise alignment distances. These models were then searched against the October 2000 Fixed Release

(with its subsequent corrections) and the April 2001 Fixed Release for data shown in Tables I and π, respectively, of the Santa Cruz contigs using the Paracel GeneMatcherf Hardware Accelerator with the Gene Wise algorithm. The sequences were chopped at 100 Kb with an overlap of 1 Kb. Each one of the superfamilies required between 3 and 6 days to complete and generate results. The results represent the coding regions of the complete final protein as it is found in the organism.

The cross-validation of the results was done in two steps. First, all of the hits

-8 with an E-value lower than 10 that did not overlap with one another were selected, and in the event of overlapping, the one with lowest E-value was selected. After selecting all of those matches, the DNA sequences were compared against the RefSeq database (Pruitt et al, Trends Genet. 16(1): 44-47, 2000) using BLAST (Altschul et al, Nucleic Acids Res. 25:3389-3402, 1997).

Over 80% of the sequences were 90% or more identical to an existing human RefSeq entry and/or mRNA from GenBank. The differences are usually due to picking the wrong model for a certain sequence that appears as a hit more than once in different families, being a different valid splice variant, which can be tested by comparing to the EST database, or by addition of a small last exon to complete the match instead of accept an stop codon in a previous one. In all of such cases, the results are easily and quickly corrected by eye. Very rarely the algorithm will actually make a wrong prediction, which is consistent with the expected behaviour (Guigo et al, Genome Res. 10(10): 1631-42, 2000). Of the remaining sequences, over 50% have a match over 90% identical in the public domain protein databases, and the differences between those sequences in the databases and the potential translations is basically the same as the differences between the DNA sequences and the RefSeq entries. The full sequences of the GPCR genes and splice variants are shown in Tables

I and II as SEQ ED NOs:l-124 (odd numbers). The amino acids encoded by these nucleic acids are shown in Tables I and II as SEQ ID NOs:l-124 (even numbers). A number of the genes were linked to markers known to be associated with human diseases genes. These are shown in Table IV. The diseases were linked to the HMM genes in the following manner: (1) the HMM gene models were compared to the consensus of the human genome sequence, located and the results kept in a relational database; (2) all possible markers (Sequence Tagged Sites (STS's)) (public or deCODE Genetics) are also located in the same consensus using ePCR or BLAT and results kept in a relational database; and (3) LOD scores for diseases are linked to markers. A span of one LOD drop around the marker was also given. A computer program takes each LOD peak and links it to the consensus through the markerhit in the database. The database is then queried for all HMM genes within the span of one LOD drop or a minimum of 15 Mb in each direction from the marker. The output is the name of the peak marker and its distance to the HMM gene.

NUCLEIC ACIDS OF THE INVENTION

Accordingly, the invention pertains to isolated nucleic acid molecules comprising human GPCR genes. The term, "GPCR", as used herein, refers to an isolated nucleic acid molecule selected from the group shown in Tables I and II, and consisting of SEQ ID NOs: 1-124 (odd numbers), and also to a portion or fragment of the isolated nucleic acid molecule (e.g., cDNA or the gene) that encodes GPCR polypeptide (e.g., a polypeptide selected from the group shown in Tables I and II, and consisting of SEQ ID NOs: 1-124 (even numbers)). In a preferred embodiment, the isolated nucleic acid molecule comprises a nucleic acid molecule selected from the group consisting of SEQ ID NOs: 1-124 (odd numbers) or the complement of such a nucleic acid molecule.

The isolated nucleic acid molecules of the present invention can be RNA, for example, mRNA, or DNA, such as cDNA and genomic DNA. DNA molecules can be double-stranded or single-stranded; single stranded RNA or DNA can be the coding, or sense, strand or the non-coding, or antisense, strand. The nucleic acid molecule can include all or a portion of the coding sequence of the gene and can further comprise additional non-coding sequences such as introns and non-coding 3' and 5' sequences (including regulatory sequences, for example). Additionally, the nucleic acid molecule can be fused to a marker sequence, for example, a sequence that encodes a polypeptide to assist in isolation or purification of the polypeptide. Such sequences include, but are not limited to, those that encode a glutathione-S- transferase (GST) fusion protein and those that encode a hemagglutinin A (HA) polypeptide marker from influenza.

An "isolated" nucleic acid molecule, as used herein, is one that is separated from nucleic acids that normally flank the gene or nucleotide sequence (as in genomic sequences) and/or has been completely or partially purified from other transcribed sequences (e.g., as in an RNA library). For example, an isolated nucleic acid of the invention may be substantially isolated with respect to the complex cellular milieu in which it naturally occurs, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized. In some instances, the isolated material will form part of a composition (for example, a crude extract containing other substances), buffer system or reagent mix. In other circumstances, the material may be purified to essential homogeneity, for example as determined by PAGE or column chromatography such as HPLC. Preferably, an isolated nucleic acid molecule comprises at least about 50, 80 or 90% (on a molar basis) of all macromolecular species present. With regard to genomic DNA, the term "isolated" also can refer to nucleic acid molecules that are separated from the chromosome with which the genomic DNA is naturally associated. For example, the isolated nucleic acid molecule can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotides which flank the nucleic acid molecule in the genomic DNA of the cell from which the nucleic acid molecule is derived. The nucleic acid molecule can be fused to other coding or regulatory sequences and still be considered isolated. Thus, recombinant DNA contained in a vector is included in the definition of "isolated" as used herein. Also, isolated nucleic acid molecules include recombinant DNA molecules in heterologous host cells, as well as partially or substantially purified DNA molecules in solution. "Isolated" nucleic acid molecules also encompass in vivo and in vitro RNA transcripts of the DNA molecules of the present invention. An isolated nucleic acid molecule or nucleotide sequence can include a nucleic acid molecule or nucleotide sequence that is synthesized chemically or by recombinant means. Therefore, recombinant DNA contained in a vector is included in the definition of "isolated" as used herein. Also, isolated nucleotide sequences include recombinant DNA molecules in heterologous organisms, as well as partially or substantially purified DNA molecules in solution. In vivo and in vitro RNA transcripts of the DNA molecules of the present invention are also encompassed by "isolated" nucleotide sequences. Such isolated nucleotide sequences are useful in the manufacture of the encoded polypeptide, as probes for isolating homologous sequences (e.g., from other mammalian species), for gene mapping (e.g., by in situ hybridization with chromosomes), or for detecting expression of the gene in tissue (e.g., human tissue), such as by Northern blot analysis.

The present invention also pertains to nucleic acid molecules which are not necessarily found in nature but which encode a GPCR polypeptide (e.g., a polypeptide having an amino acid sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:l-124 (even numbers)), or another splicing variant of a GPCR polypeptide or polymoφhic variant thereof.

Thus, for example, DNA molecules which comprise a sequence that is different from the naturally-occurring nucleotide sequence but which, due to the degeneracy of the genetic code, encode a GPCR polypeptide of the present invention are also the subject of this invention. The invention also encompasses nucleotide sequences encoding portions (fragments), or encoding variant polypeptides such as analogues or derivatives of a GPCR polypeptide. Such variants can be naturally-occurring, such as in the case of allelic variation or single nucleotide polymorphisms, or non- naturally-occurring, such as those induced by various mutagens and mutagenic processes. Intended variations include, but are not limited to, addition, deletion and substitution of one or more nucleotides that can result in conservative or non- conservative amino acid changes, including additions and deletions. Preferably the nucleotide (and/or resultant amino acid) changes are silent or conserved; that is, they do not alter the characteristics or activity of a GPCR polypeptide. In one preferred embodiment, the nucleotide sequences are fragments that comprise one or more polymorphic microsatellite markers. In another preferred embodiment, the nucleotide sequences are fragments that comprise one or more single nucleotide polymorphisms in a GPCR gene.

Other alterations of the nucleic acid molecules of the invention can include, for example, labeling, methylation, internucleotide modifications such as uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoamidates, carbamates), charged linkages (e.g., phosphorothioates, phosphorodithioates), pendent moieties (e.g., polypeptides), intercalators (e.g., acridine, psoralen), chelators, alkylators, and modified linkages (e.g., alpha anomeric nucleic acids). Also included are synthetic molecules that mimic nucleic acid molecules in the ability to bind to designated sequences via hydrogen bonding and other chemical interactions. Such molecules include, for example, those in which peptide linkages substitute for phosphate linkages in the backbone of the molecule. The invention also pertains to nucleic acid molecules that hybridize under high stringency hybridization conditions, such as for selective hybridization, to a nucleotide sequence described herein (e.g., nucleic acid molecules which specifically hybridize to a nucleotide sequence encoding polypeptides described herein, and, optionally, have an activity of the polypeptide). In one embodiment, the invention includes variants described herein which hybridize under high stringency hybridization conditions (e.g., for selective hybridization) to a nucleotide sequence comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-124 (odd numbers). In another embodiment, the invention includes variants described herein which hybridize under high stringency hybridization conditions

(e.g., for selective hybridization) to a nucleotide sequence encoding an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-124 (even numbers) or a polymorphic variant thereof. In a preferred embodiment, the variant that hybridizes under high stringency hybridizations has an activity of a GPCR. Such nucleic acid molecules can be detected and/or isolated by specific hybridization (e.g., under high stringency conditions). "Specific hybridization," as used herein, refers to the ability of a first nucleic acid to hybridize to a second nucleic acid in a manner such that the first nucleic acid does not hybridize to any nucleic acid other than to the second nucleic acid (e.g., when the first nucleic acid has a higher similarity to the second nucleic acid than to any other nucleic acid in a sample wherein the hybridization is to be performed). "Stringency conditions" for hybridization is a term of art which refers to the incubation and wash conditions, e.g., conditions of temperature and buffer concentration, which permit hybridization of a particular nucleic acid to a second nucleic acid; the first nucleic acid may be perfectly (i.e., 100%) complementary to the second, or the first and second may share some degree of complementarity which is less than perfect (e.g., 70%, 75%, 85%>, 90%, 95%). For example, certain high stringency conditions can be used which distinguish perfectly complementary nucleic acids from those of less complementarity. "High stringency conditions", "moderate stringency conditions" and "low stringency conditions" for nucleic acid hybridizations are explained on pages 2.10.1-2.10.16 and pages 6.3.1-6.3.6 in Current Protocols in Molecular Biology (Ausubel, F.M. et al, "Current Protocols in Molecular Biology", John Wiley & Sons, 1998 and 2001), the entire teachings of which are incorporated by reference herein). The exact conditions which determine the stringency of hybridization depend not only on ionic strength (e.g., 0.2X SSC, 0. IX SSC), temperature (e.g., room temperature, 42°C, 68°C) and the concentration of destabilizing agents such as formamide or denaturing agents such as SDS, but also on factors such as the length of the nucleic acid sequence, base composition, percent mismatch between hybridizing sequences and the frequency of occurrence of subsets of that sequence within other non-identical sequences. Thus, equivalent conditions can be determined by varying one or more of these parameters while maintaining a similar degree of identity or similarity between the two nucleic acid molecules. Typically, conditions are used such that sequences at least about 60%, at least about

70%, at least about 80%, at least about 90%) or at least about 95% or more identical to each other remain hybridized to one another. By varying hybridization conditions from a level of stringency at which no hybridization occurs to a level at which hybridization is first observed, conditions which will allow a given sequence to hybridize (e.g., selectively) with the most similar sequences in the sample can be determined.

Exemplary conditions are described in Krause, M.H. and S.A. Aaronson, Methods in Enzymology 200:546-556, 1991, and in, Ausubel, et al, "Current Protocols in Molecular Biology", John Wiley & Sons, 1998 and 2001, which describes the determination of washing conditions for moderate or low stringency conditions. Washing is the step in which conditions are usually set so as to determine a minimum level of complementarity of the hybrids. Generally, starting from the lowest temperature at which only homologous hybridization occurs, each °C by which the final wash temperature is reduced (holding SSC concentration constant) allows an increase by 1% in the maximum extent of mismatching among the sequences that hybridize. Generally, doubling the concentration of SSC results in an increase in T_m of -17°C. Using these guidelines, the washing temperature can be determined empirically for high, moderate or low stringency, depending on the level of mismatch sought. For example, a low stringency wash can comprise washing in a solution containing 0.2X SSC/0.1% SDS for 10 minutes at room temperature; a moderate stringency wash can comprise washing in a prewarmed solution (42°C) solution containing 0.2X SSC/0.1% SDS for 15 minutes at 42°C; and a high stringency wash can comprise washing in prewarmed (68°C) solution containing 0.1X SSC/0.1%SDS for 15 minutes at 68°C. Furthermore, washes can be performed repeatedly or sequentially to obtain a desired result as known in the art. Equivalent conditions can be determined by varying one or more of the parameters given as an example, as known in the art, while maintaining a similar degree of identity or similarity between the target nucleic acid molecule and the primer or probe used. The percent identity of two nucleotide or amino acid sequences can be determined by aligning the sequences for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first sequence). The nucleotides or amino acids at corresponding positions are then compared, and the percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity = # of identical positions/total # of positions x 100). In certain embodiments, the length of a sequence aligned for comparison puφoses is at least 30%, preferably at least 40%, more preferably at least 60%, and even more preferably at least 70%, 80%, 90% or 95% of the length of the reference sequence.

The actual comparison of the two sequences can be accomplished by well-known methods, for example, using a mathematical algorithm. A preferred, non-limiting example of such a mathematical algorithm is described in Karlin et al, Proc. Natl. Acad. Sci. USA 90:5873-5877, 1993. Such an algorithm is incoφorated into the NBLAST and XBLAST programs (version 2.0) as described in Altschul et al. ,

Nucleic Acids Res. 25:389-3402, 1997. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., NBLAST) can be used. In one embodiment, parameters for sequence comparison can be set at score=100, wordlength=12, or can be varied (e.g., W=5 or W=20). Another preferred, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller C_3/OS 4(1): 11-17, 1998. Such an algorithm is incoφorated into the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package (ACCELRYS, Cambridge, UK). When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used. Additional algorithms for sequence analysis are known in the art and include ADVANCE and ADAM as described in Torellis and Robotti, Comput. Appl. Biosci. 10:3-5, 1994; and FASTA described in Pearson and Lipman Proc. Natl. Acad. Sci. USA 85:2444-8, 1988. In another embodiment, the percent identity between two amino acid sequences can be accomplished using the GAP program in the GCG software package using either a BLOSUM63 matrix or a PAM250 matrix, and a gap weight of 12, 10, 8, 6, or 4 and a length weight of 2, 3, or 4. In yet another embodiment, the percent identity between two nucleic acid sequences can be accomplished using the GAP program in the GCG software package, using a gap weight of 50 and a length weight of 3.

The present invention also provides isolated nucleic acid molecules that contain a fragment or portion that hybridizes under highly stringent conditions to a nucleotide sequence comprising a nucleotide sequence selected from the group consisting of SEQ JD NOs: 1 - 124 (odd numbers), or the complement of such a sequence, and also provides isolated nucleic acid molecules that contain a fragment or portion that hybridizes under highly stringent conditions to a nucleotide sequence encoding an amino acid sequence selected SEQ ID NOs: 1-124 (even numbers), or polymoφhic variant thereof. The nucleic acid fragments of the invention are at least about 15, preferably at least about 18, 20, 23 or 25 nucleotides, and can be 30, 40, 50, 100, 200 or more nucleotides in length. Longer fragments, for example, 30 or more nucleotides in length, which encode antigenic polypeptides described herein are particularly useful, such as for the generation of antibodies as described below.

In a related aspect, the nucleic acid fragments of the invention are used as probes or primers in assays such as those described herein. "Probes" or "primers" are oligonucleotides that hybridize in a base-specific manner to a complementary strand of nucleic acid molecules. Such probes and primers include polypeptide nucleic acids, as described in Nielsen et al, Science 254: 1497-1500, (1991).

Typically, a probe or primer comprises a region of nucleotide sequence that hybridizes to at least about 15, typically about 20-25, and more typically about 40, 50 or 75, consecutive nucleotides of a nucleic acid molecule comprising a contiguous nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-124 (odd numbers), or the complement of such a sequence, or a sequence encoding an amino acid sequence selected from SEQ ID NOs: 1-124 (even numbers), or polymoφhic variant thereof. In preferred embodiments, a probe or primer comprises 100 or fewer nucleotides, preferably from 6 to 50 nucleotides, preferably from 12 to 30 nucleotides. In other embodiments, the probe or primer is at least 70%) identical to the contiguous nucleotide sequence or to the complement of the contiguous nucleotide sequence, preferably at least 80% identical, more preferably at least 90% identical, even more preferably at least 95% identical, or even capable of selectively hybridizing to the contiguous nucleotide sequence or to the complement of the contiguous nucleotide sequence. Often, the probe or primer further comprises a label, e.g., radioisotope, fluorescent compound, enzyme, or enzyme co-factor.

The nucleic acid molecules of the invention such as those described above can be identified and isolated using standard molecular biology techniques and the sequence information provided herein. For example, nucleic acid molecules can be amplified and isolated by the polymerase chain reaction using synthetic oligonucleotide primers designed based on one or more of the sequences selected from the group consisting of SEQ ID NOs: 1-124 (odd numbers), or the complement of such a sequence, or designed based on nucleotides based on sequences encoding one or more of the amino acid sequences provided herein. See generally PCR Technology: Principles and Applications for DNA Amplification (ed. H.A. Erlich,

Freeman Press, NY, NY, 1992); PCR Protocols: A Guide to Methods and Applications (Eds. Innis et al, Academic Press, San Diego, CA, 1990); Mattila et al., Nucl Acids Res. 19:4967, 1991; Eckert et al, PCR Methods and Applications 1 : 17, 1991 ; PCR (eds. McPherson et al, IRL Press, Oxford); and U.S. Patent 4,683,202. The nucleic acid molecules can be amplified using cDNA, mRNA or genomic DNA as a template, cloned into an appropriate vector and characterized by DNA sequence analysis. Other suitable amplification methods include the ligase chain reaction (LCR)

(see Wu and Wallace, Genomics 4:560, 1989, Landegren et al, Science 241 : 1077, 1988, transcription amplification (Kwoh et al, Proc. Natl. Acad. Sci. USA 86:1 173, 1989), and self-sustained sequence replication (Guatelli et al, Proc. Nat. Acad. Sci. USA 87: 1874, 1990) and nucleic acid based sequence amplification (NASBA). The latter two amplification methods involve isothermal reactions based on isothermal transcription, which produce both single stranded RNA (ssRNA) and double stranded DNA (dsDNA) as the amplification products in a ratio of about 30 or 100 to 1, respectively.

The amplified DNA can be radiolabelled and used as a probe for screening a cDNA library derived from human cells, mRNA in zap express, ZIPLOX or other suitable vector. Corresponding clones can be isolated, DNA can obtained following in vivo excision, and the cloned insert can be sequenced in either or both orientations by art recognized methods to identify the correct reading frame encoding a polypeptide of the appropriate molecular weight. For example, the direct analysis of the nucleotide sequence of nucleic acid molecules of the present invention can be accomplished using well-known methods that are commercially available. See, for example, Sambrook et al, Molecular Cloning, A Laboratory Manual (2nd Ed., CSHP, New York 1989); Zyskind et al, Recombinant DNA Laboratory Manual, (Acad. Press, 1988)). Using these or similar methods, the polypeptide and the DNA encoding the polypeptide can be isolated, sequenced and further characterized.

Antisense nucleic acid molecules of the invention can be designed using the nucleotide sequences of one or more of SEQ ID NOs: 1-124 (odd numbers) and or the complement of one or more of SEQ ID NOs: 1-124 (odd numbers), and/or a portion of one or more of SEQ ID NOs: 1-124 (odd numbers), or the complement of one or more of SEQ ID NOs:l-124 (odd numbers) and/or a sequence encoding the amino acid sequences of one or more of SEQ ID NOs: 1-124 (even numbers), or encoding a portion of one or more of SEQ ID NOs: 1-124 (even numbers), and constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid molecule (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used. Alternatively, the antisense nucleic acid molecule can be produced biologically using an expression vector into which a nucleic acid molecule has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid molecule will be of an antisense orientation to a target nucleic acid of interest).

In general, the isolated nucleic acid sequences of the invention can be used as molecular weight markers on Southern gels, and as chromosome markers which are labeled to map related gene positions. The nucleic acid sequences can also be used to compare with endogenous DNA sequences in patients to identify one or more of the disorders described above, and as probes, such as to hybridize and discover related DNA sequences or to subtract out known sequences from a sample. The nucleic acid sequences can further be used to derive primers for genetic fingeφrinting, to raise anti-polypeptide antibodies using DNA immunization techniques, and as an antigen to raise anti-DNA antibodies or elicit immune responses. Portions or fragments of the nucleotide sequences identified herein (and the corresponding complete gene sequences) can be used in numerous ways as polynucleotide reagents. For example, these sequences can be used to: (i) map their respective genes on a chromosome; and, thus, locate gene regions associated with genetic disease; (ii) identify an individual from a minute biological sample (tissue typing); and (iii) aid in forensic identification of a biological sample. Additionally, the nucleotide sequences of the invention can be used to identify and express recombinant polypeptides for analysis, characterization or therapeutic use, or as markers for tissues in which the corresponding polypeptide is expressed, either constitutively, during tissue differentiation, or in diseased states. The nucleic acid sequences can additionally be used as reagents in the screening and/or diagnostic assays described herein, and can also be included as components of kits (e.g., reagent kits) for use in the screening and/or diagnostic assays described herein.

Another aspect of the invention pertains to nucleic acid constructs containing a nucleic acid molecule selected from the group consisting of SEQ ID NOs: 1-124 (odd numbers) and the complements thereof (or a portion thereof). Yet another aspect of the invention pertains to nucleic acid constructs containing a nucleic acid molecule encoding an amino acid sequence of SEQ ID NOs: 1-124 (even numbers) or polymoφhic variant thereof. The constructs comprise a vector (e.g., an expression vector) into which a sequence of the invention has been inserted in a sense or antisense orientation. As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid", which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors, expression vectors, are capable of directing the expression of genes to which they are operably linked. In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses) that serve equivalent functions.

Preferred recombinant expression vectors of the invention comprise a nucleic acid molecule of the invention in a form suitable for expression of the nucleic acid molecule in a host cell. This means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, which is operably linked to the nucleic acid sequence to be expressed. Within a recombinant expression vector, "operably linked" or "operatively linked" is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner which allows for expression of the nucleotide sequence (e.g., in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell). The term "regulatory sequence" is intended to include promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are described, for example, in Goeddel, "Gene Expression Technology", Methods in Enzymology

185, Academic Press, San Diego, CA (1990). Regulatory sequences include those which direct constitutive expression of a nucleotide sequence in many types of host cell and those which direct expression of the nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences). It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed and the level of expression of polypeptide desired. The expression vectors of the invention can be introduced into host cells to thereby produce polypeptides, including fusion polypeptides, encoded by nucleic acid molecules as described herein. The recombinant expression vectors of the invention can be designed for expression of a polypeptide of the invention in prokaryotic or eukaryotic cells, e.g., bacterial cells such as E. coli, insect cells (using baculovirus expression vectors), yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, supra. Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.

Another aspect of the invention pertains to host cells into which a recombinant expression vector of the invention has been introduced. The terms "host cell" and

"recombinant host cell" are used interchangeably herein. It is understood that such terms refer not only to the particular subject cell but also to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.

A host cell can be any prokaryotic or eukaryotic cell. For example, a nucleic acid molecule of the invention can be expressed in bacterial cells (e.g., E. coli), insect cells, yeast or mammalian cells (such as Chinese hamster ovary cells (CHO) or COS cells). Other suitable host cells are known to those skilled in the art.

Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms "transformation" and "transfection" are intended to refer to a variety of art- recognized techniques for introducing a foreign nucleic acid molecule (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co-precipitation,

DEAE-dextran-mediated transfection, lipofection, or electroporation. Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (supra), and other laboratory manuals.

For stable transfection of mammalian cells, it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome. In order to identify and select these integrants, a gene that encodes a selectable marker (e.g., for resistance to antibiotics) is generally introduced into the host cells along with the gene of interest. Preferred selectable markers include those that confer resistance to drugs, such as G418, hygromycin and methotrexate. Nucleic acid molecules encoding a selectable marker can be introduced into a host cell on the same vector as the nucleic acid molecule of the invention or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid molecule can be identified by drug selection (e.g., cells that have incoφorated the selectable marker gene will survive, while the other cells die).

A host cell of the invention, such as a prokaryotic or eukaryotic host cell in culture, can be used to produce (i.e., express) a polypeptide of the invention. Accordingly, the invention further provides methods for producing a polypeptide using the host cells of the invention. In one embodiment, the method comprises culturing the host cell of invention (into which a recombinant expression vector encoding a polypeptide of the invention has been introduced) in a suitable medium such that the polypeptide is produced. In another embodiment, the method further comprises isolating the polypeptide from the medium or the host cell.

The host cells of the invention can also be used to produce nonhuman transgenic animals. For example, in one embodiment, a host cell of the invention is a fertilized oocyte or an embryonic stem cell into which a nucleic acid molecule of the invention has been introduced (e.g., an exogenous GPCR gene, or an exogenous nucleic acid encoding a GPCR polypeptide). Such host cells can then be used to create non-human transgenic animals in which exogenous nucleotide sequences have been introduced into the genome or homologous recombinant animals in which endogenous nucleotide sequences have been altered. Such animals are useful for studying the function and/or activity of the nucleotide sequence and polypeptide encoded by the sequence and for identifying and/or evaluating modulators of their activity. As used herein, a "transgenic animal" is a non-human animal, preferably a mammal, more preferably a rodent such as a rat or mouse, in which one or more of the cells of the animal include a transgene. Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens and amphibians. A transgene is exogenous DNA which is integrated into the genome of a cell from which a transgenic animal develops and which remains in the genome of the mature animal, thereby directing the expression of an encoded gene product in one or more cell types or tissues of the transgenic animal. As used herein, an "homologous recombinant animal" is a non-human animal, preferably a mammal, more preferably a mouse, in which an endogenous gene has been altered by homologous recombination between the endogenous gene and an exogenous DNA molecule introduced into a cell of the animal, e.g., an embryonic cell of the animal, prior to development of the animal.

Methods for generating transgenic animals via embryo manipulation and microinjection, particularly animals such as mice, have become conventional in the art and are described, for example, in U.S. Patent Nos. 4,736,866 and 4,870,009, U.S. Pat. No. 4,873,191 and in Hogan, Manipulating the Mouse Embryo (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986). Methods for constructing homologous recombination vectors and homologous recombinant animals are described further in Bradley (1991) Current Opinion in BioTechnology

2:823-829 and in PCT Publication Nos. WO 90/11354, WO 91/01140, WO 92/0968, and WO 93/04169. Clones of the non-human transgenic animals described herein can also be produced according to the methods described in Wilmut et al (1997) Nature 385:810-813 and PCT Publication Nos. WO 97/07668 and WO 97/07669.

POLYPEPTIDES OF THE INVENTION The present invention also pertains to isolated polypeptides encoded by

GPCRs ("GPCR polypeptides") and fragments and variants thereof, as well as polypeptides encoded by nucleotide sequences described herein (e.g., other splicing variants). The term "polypeptide" refers to a polymer of amino acids, and not to a specific length; thus, peptides, oligopeptides and proteins are included within the definition of a polypeptide. As used herein, a polypeptide is said to be "isolated" or

"purified" when it is substantially free of cellular material when it is isolated from recombinant and non-recombinant cells, or free of chemical precursors or other chemicals when it is chemically synthesized. A polypeptide, however, can be joined to another polypeptide with which it is not normally associated in a cell (e.g., in a "fusion protein") and still be "isolated" or "purified."

The polypeptides of the invention can be purified to homogeneity. It is understood, however, that preparations in which the polypeptide is not purified to homogeneity are useful. The critical feature is that the preparation allows for the desired function of the polypeptide, even in the presence of considerable amounts of other components. Thus, the invention encompasses various degrees of purity. In one embodiment, the language "substantially free of cellular material" includes preparations of the polypeptide having less than about 30% (by dry weight) other proteins (i.e., contaminating protein), less than about 20% other proteins, less than about 10% other proteins, or less than about 5% other proteins. When a polypeptide is recombinantly produced, it can also be substantially free of culture medium, i.e., culture medium represents less than about 20%, less than about 10%, or less than about 5% of the volume of the polypeptide preparation. The language "substantially free of chemical precursors or other chemicals" includes preparations of the polypeptide in which it is separated from chemical precursors or other chemicals that are involved in its synthesis. In one embodiment, the language

"substantially free of chemical precursors or other chemicals" includes preparations of the polypeptide having less than about 30%ι (by dry weight) chemical precursors or other chemicals, less than about 20% chemical precursors or other chemicals, less than about 10%) chemical precursors or other chemicals, or less than about 5% chemical precursors or other chemicals.

In one embodiment, a polypeptide of the invention comprises an amino acid sequence encoded by a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-124 (odd numbers), or the complement of such a nucleic acid, or portions thereof, e.g., SEQ ID NO: 1-124 (even numbers), or a portion or polymoφhic variant thereof. However, the polypeptides of the invention also encompass fragment and sequence variants. Variants include a substantially homologous polypeptide encoded by the same genetic locus in an organism, i.e., an allelic variant, as well as other splicing variants. Variants also encompass polypeptides derived from other genetic loci in an organism, but having substantial homology to a polypeptide encoded by a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-124 (odd numbers), or a complement of such a sequence, or portions thereof, or having substantial homology to a polypeptide encoded by a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of nucleotide sequences encoding SEQ ID NOs:l-124 (even numbers), or polymoφhic variants thereof. Variants also include polypeptides substantially homologous or identical to these polypeptides but derived from another organism, i.e., an ortholog. Variants also include polypeptides that are substantially homologous or identical to these polypeptides that are produced by chemical synthesis. Variants also include polypeptides that are substantially homologous or identical to these polypeptides mat are produced by recombinant methods. As used herein, two polypeptides (or a region of the polypeptides) are substantially homologous or identical when the amino acid sequences are at least about 45-55%), typically at least about 70-75%, more typically at least about 80- 85%, and most typically greater than about 90% or more homologous or identical. A substantially homologous amino acid sequence, according to the present invention, will be encoded by a nucleic acid molecule hybridizing to one or more of SEQ ID NOs: 1-124 (odd numbers), or portion thereof, under stringent conditions as more particularly described above, or will be encoded by a nucleic acid molecule hybridizing to a nucleic acid sequence encoding one of SEQ ID NOs: 1-124 (even numbers), a portion thereof or polymoφhic variant thereof, under stringent conditions as more particularly described thereof. To determine the percent homology or identity of two amino acid sequences, or of two nucleic acid sequences, the sequences are aligned for optimal comparison puφoses (e.g., gaps can be introduced in the sequence of one polypeptide or nucleic acid molecule for optimal alignment with the other polypeptide or nucleic acid molecule). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in one sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the other sequence, then the molecules are homologous at that position. As used herein, amino acid or nucleic acid "homology" is equivalent to amino acid or nucleic acid "identity". The percent homology between the two sequences is a function of the number of identical positions shared by the sequences (i.e., percent homology equals the number of identical positions/total number of positions times 100). The invention also encompasses polypeptides having a lower degree of identity but having sufficient similarity so as to perform one or more of the same functions performed by a polypeptide encoded by a nucleic acid molecule of the invention. Similarity is determined by conserved amino acid substitution. Such substitutions are those that substitute a given amino acid in a polypeptide by another amino acid of like characteristics. Conservative substitutions are likely to be phenotypically silent. Typically seen as conservative substitutions are the replacements, one for another, among the aliphatic amino acids Ala, Val, Leu and He; interchange of the hydroxyl residues Ser and Thr, exchange of the acidic residues Asp and Glu, substitution between the amide residues Asn and Gin, exchange of the basic residues Lys and Arg and replacements among the aromatic residues Phe and Tyr. Guidance concerning which amino acid changes are likely to be phenotypically silent are found in Bowie et al, Science 247:1306-1310 (1990).

A variant polypeptide can differ in amino acid sequence by one or more substitutions, deletions, insertions, inversions, fusions, and truncations or a combination of any of these. Further, variant polypeptides can be fully functional or can lack function in one or more activities. Fully functional variants typically contain only conservative variation or variation in non-critical residues or in non- critical regions. Functional variants can also contain substitution of similar amino acids that result in no change or an insignificant change in function. Alternatively, such substitutions may positively or negatively affect function to some degree.

Non-functional variants typically contain one or more non-conservative amino acid substitutions, deletions, insertions, inversions, or truncation or a substitution, insertion, inversion, or deletion in a critical residue or critical region.

Amino acids that are essential for function can be identified by methods known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis

(Cunningham et al, Science 244:1081-1085 (1989)). The latter procedure introduces single alanine mutations at every residue in the molecule. The resulting mutant molecules are then tested for biological activity in vitro, or in vitro proliferative activity. Sites that are critical for polypeptide activity can also be determined by structural analysis such as crystallization, nuclear magnetic resonance or photoaffinity labeling (Smith et al, J. Mol. Biol. 224:899-904 (1992); de Vos et al, Science 255:306-312 (1992)). The invention also includes polypeptide fragments of the polypeptides of the invention. Fragments can be derived from a polypeptide encoded by a nucleic acid molecule comprising one of SEQ ID NOs:l-124 (odd numbers), or a complement of such a nucleic acid (e.g., SEQ ID NOs:l-124 (even numbers), or other variants). However, the invention also encompasses fragments of the variants of the polypeptides described herein. As used herein, a fragment comprises at least 6 contiguous amino acids. Useful fragments include those that retain one or more of the biological activities of the polypeptide as well as fragments that can be used as an immunogen to generate polypeptide-specific antibodies. Biologically active fragments (peptides which are, for example, 6, 9, 12, 15,

16, 20, 30, 35, 36, 37, 38, 39, 40, 50, 100 or more amino acids in length) can comprise a domain, segment, or motif that has been identified by analysis of the polypeptide sequence using well-known methods, e.g., signal peptides, extracellular domains, one or more transmembrane segments or loops, ligand binding regions, zinc finger domains, DNA binding domains, acylation sites, glycosylation sites, or phosphorylation sites.

Fragments can be discrete (not fused to other amino acids or polypeptides) or can be within a larger polypeptide. Further, several fragments can be comprised within a single larger polypeptide. In one embodiment a fragment designed for expression in a host can have heterologous pre- and pro-polypeptide regions fused to the amino terminus of the polypeptide fragment and an additional region fused to the carboxyl terminus of the fragment.

The invention thus provides chimeric or fusion polypeptides. These comprise a polypeptide of the invention operatively linked to a heterologous protein or polypeptide having an amino acid sequence not substantially homologous to the polypeptide. "Operatively linked" indicates that the polypeptide and the heterologous protein are fused in-frame. The heterologous protein can be fused to the N-terminus or C-terminus of the polypeptide. In one embodiment the fusion polypeptide does not affect function of the polypeptide per se. For example, the fusion polypeptide can be a GST-fusion polypeptide in which the polypeptide sequences are fused to the C-terminus of the GST sequences. Other types of fusion polypeptides include, but are not limited to, enzymatic fusion polypeptides, for example β-galactosidase fusions, yeast two-hybrid GAL fusions, poly-His fusions and Ig fusions. Such fusion polypeptides, particularly poly-His fusions, can facilitate the purification of recombinant polypeptide. In certain host cells (e.g. , mammalian host cells), expression and/or secretion of a polypeptide can be increased using a heterologous signal sequence. Therefore, in another embodiment, the fusion polypeptide contains a heterologous signal sequence at its N-terminus. EP-A-O 464 533 discloses fusion proteins comprising various portions of immunoglobulin constant regions. The Fc is useful in therapy and diagnosis and thus results, for example, in improved pharmacokinetic properties (EP-A 0232 262). In drug discovery, for example, human proteins have been fused with Fc portions for the puφose of high-throughput screening assays to identify antagonists. Bennett et al, J. Mol. Recog. 8:52-58 (1995) and Johanson et al, J. Biol. Chem. 270:16:9459- 9471 (1995). Thus, this invention also encompasses soluble fusion polypeptides containing a polypeptide of the invention and various portions of the constant regions of heavy or light chains of immunoglobulins of various subclasses (IgG, IgM, IgA, IgE).

A chimeric or fusion polypeptide can be produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different polypeptide sequences are ligated together in-frame in accordance with conventional techniques. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of nucleic acid fragments can be carried out using anchor primers which give rise to complementary overhangs between two consecutive nucleic acid fragments which can subsequently be annealed and re-amplified to generate a chimeric nucleic acid sequence (see Ausubel et al, Current Protocols in Molecular Biology, 1992). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST protein). A nucleic acid molecule encoding a polypeptide of the invention can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the polypeptide.

The isolated polypeptide can be purified from cells that naturally express it, purified from cells that have been altered to express it (recombinant), or synthesized using known protein synthesis methods. In one embodiment, the polypeptide is produced by recombinant DNA techniques. For example, a nucleic acid molecule encoding the polypeptide is cloned into an expression vector, the expression vector introduced into a host cell and the polypeptide expressed in the host cell. The polypeptide can then be isolated from the cells by an appropriate purification scheme using standard protein purification techniques.

In general, polypeptides of the present invention can be used as a molecular weight marker on SDS-PAGE gels or on molecular sieve gel filtration columns using art-recognized methods. The polypeptides of the present invention can be used to raise antibodies or to elicit an immune response. The polypeptides can also be used as a reagent, e.g., a labeled reagent, in assays to quantitatively determine levels of the polypeptide or a molecule to which it binds (e.g., a ligand) in biological fluids. The polypeptides can also be used as markers for cells or tissues in which the corresponding polypeptide is preferentially expressed, either constitutively, during tissue differentiation, or in a disease state. The polypeptides can be used to isolate a corresponding binding agent, e.g., ligand, such as, for example, in an interaction trap assay, and to screen for peptide or small molecule antagonists or agonists of the binding interaction.

ANTIBODIES OF THE INVENTION

Polyclonal and/or monoclonal antibodies that specifically bind one form of the gene product but not to the other form of the gene product are also provided. Antibodies are also provided which bind a portion of either the variant or the reference gene product that contains the polymoφhic site or sites. The invention provides antibodies to the polypeptides and polypeptide fragments of the invention, e.g., having an amino acid sequence of one of SEQ ID NOs: 1-124 (even numbers) or a portion thereof, or having an amino acid sequence encoded by a nucleic acid molecule comprising all or a portion of SEQ JJD NOs: 1-124 (odd numbers), or a complement or another variant or portion thereof. The term "antibody" as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site that specifically binds an antigen. A molecule that specifically binds to a polypeptide of the invention is a molecule that binds to that polypeptide or a fragment thereof, but does not substantially bind other molecules in a sample, e.g., a biological sample, which naturally contains the polypeptide. Examples of immunologically active portions of immunoglobulin molecules include F(ab) and F(ab')₂ fragments which can be generated by treating the antibody with an enzyme such as pepsin. The invention provides polyclonal and monoclonal antibodies that bind to a polypeptide of the invention. The term "monoclonal antibody" or "monoclonal antibody composition", as used herein, refers to a population of antibody molecules that contain only one species of an antigen binding site capable of immunoreacting with a particular epitope of a polypeptide of the invention. A monoclonal antibody composition thus typically displays a single binding affinity for a particular polypeptide of the invention with which it immunoreacts.

Polyclonal antibodies can be prepared as described above by immunizing a suitable subject with a desired immunogen, e.g., polypeptide of the invention or fragment thereof. The antibody titer in the immunized subject can be monitored over time by standard techniques, such as with an enzyme linked immunosorbent assay (ELISA) using immobilized polypeptide. If desired, the antibody molecules directed against the polypeptide can be isolated from the mammal (e.g., from the blood) and further purified by well-known techniques, such as protein A chromatography to obtain the IgG fraction. At an appropriate time after immunization, e.g., when the antibody titers are highest, antibody-producing cells can be obtained from the subject and used to prepare monoclonal antibodies by standard techniques, such as the hybridoma technique originally described by Kohler and Milstein (1975) Nature 256:495-497, the human B cell hybridoma technique (Kozbor et al. (1983) Immunol. Today 4:72), The EBV-hybridoma technique (Cole et al. (1985), Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96) or trioma techniques. The technology for producing hybridomas is well known (see generally Current Protocols in Immunology (1994) Coligan et al. (Eds.) John Wiley & Sons, Inc., New York, NY). Briefly, an immortal cell line (typically a myeloma) is fused to lymphocytes (typically splenocytes) from a mammal immunized with an immunogen as described above, and the culture supernatants of the resulting hybridoma cells are screened to identify a hybridoma producing a monoclonal antibody that binds a polypeptide of the invention.

Any of the many well known protocols used for fusing lymphocytes and immortalized cell lines can be applied for the puφose of generating a monoclonal antibody to a polypeptide of the invention (see, e.g., Current Protocols in Immunology, supra; Galfre et al, Nature 266:55052 (1977); R.H. Kenneth, in Monoclonal Antibodies: A New Dimension In Biological Analyses, Plenum

Publishing Coφ., New York, New York (1980); and Lerner, YaleJ. Biol. Med. 54:387-402 (1981). Moreover, the ordinarily skilled worker will appreciate that there are many variations of such methods that also would be useful.

Alternative to preparing monoclonal antibody-secreting hybridomas, a monoclonal antibody to a polypeptide of the invention can be identified and isolated by screening a recombinant combinatorial immunoglobulin library (e.g., an antibody phage display library) with the polypeptide to thereby isolate immunoglobulin library members that bind the polypeptide. Kits for generating and screening phage display libraries are commercially available (e.g., the Pharmacia Recombinant Phage Antibody System, Catalog No. 27-9400-01 ; and the Stratagene Swr/ZAP™

Phage Display Kit, Catalog No. 240612). Additionally, examples of methods and reagents particularly amenable for use in generating and screening antibody display library can be found in, for example, U.S. Patent No. 5,223,409; PCT Publication No. WO 92/18619; PCT Publication No. WO 91/17271 ; PCT Publication No. WO 92/20791 ; PCT Publication No. WO 92/15679; PCT Publication No. WO 93/01288;

PCT Publication No. WO 92/01047; PCT Publication No. WO 92/09690; PCT Publication No. WO 90/02809; Fuchs et al, Bio/Technology 9:1370-1372 (1991); Hay et al, Hum. Antibod. Hybridomas 3:81-85 (1992); Huse et al, Science 246: 1275-1281 (1989); Griffiths et al, EMBOJ. 12:725-734 (1993).

Additionally, recombinant antibodies, such as chimeric and humanized monoclonal antibodies, comprising both human and non-human portions, which can be made using standard recombinant DNA techniques, are within the scope of the invention. Such chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art.

In general, antibodies of the invention (e.g., a monoclonal antibody) can be used to isolate a polypeptide of the invention by standard techniques, such as affinity chromatography or immunoprecipitation. A polypeptide-specific antibody can facilitate the purification of natural polypeptide from cells and of recombinantly produced polypeptide expressed in host cells. Moreover, an antibody specific for a polypeptide of the invention can be used to detect the polypeptide (e.g., in a cellular lysate, cell supernatant, or tissue sample) in order to evaluate the abundance and pattern of expression of the polypeptide. Antibodies can be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, e.g., to, for example, determine the efficacy of a given treatment regimen. Antibody detection can be facilitated by coupling it to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials.

Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable

125 131 35 3 radioactive material include I, I, S or H.

DIAGNOSTIC AND SCREENING ASSAYS OF THE INVENTION

The present invention also pertains to a method of diagnosing or aiding in the diagnosis of a disease or condition associated with a GPCR gene or gene product in an individual. Diagnostic assays can be designed for assessing GPCR gene expression, or for assessing activity of GPCR polypeptides of the invention. In one embodiment, the assays are used in the context of a biological sample (e.g., blood, serum, cells, tissue) to thereby determine whether an individual is afflicted with a disease or condition associated with a GPCR, or a defect in a GPCR. The invention also provides for prognostic (or predictive) assays for determining whether an individual is susceptible to a disease of condition associated with a GPCR, e.g., if an individual is at risk for addiction to an opoid. For example, mutations in the gene can be assayed in a biological sample. Such assays can be used for prognostic or predictive puφose to thereby prophylactically treat an individual prior to the onset of symptoms associated with a susceptibility to a disease or condition associated with a GPCR. Another aspect of the invention pertains to assays for monitoring the influence of agents (e.g., drugs, compounds or other agents) on the gene expression or activity of polypeptides of the invention, as well as to assays for identifying agents that bind to a polypeptides. These and other assays and agents are described in further detail in the following sections.

DIAGNOSTIC ASSAYS

The nucleic acids, probes, primers, polypeptides and antibodies described herein can be used in methods of diagnosis of a susceptibility to a disease or condition associated with a GPCR, as well as in kits useful for diagnosis of a susceptibility to a disease or condition associated with a GPCR.

In one embodiment of the invention, susceptibility to a disease or condition associated with a GPCR can be diagnosed by detecting a polymoφhism in a GPCR as described herein. The polymoφhism can be a mutation in a GPCR, such as the insertion or deletion of a single nucleotide, or of more than one nucleotide, resulting in a frame shift mutation; the change of at least one nucleotide, resulting in a change in the encoded amino acid; the change of at least one nucleotide, resulting in the generation of a premature stop codon; the deletion of several nucleotides, resulting in a deletion of one or more amino acids encoded by the nucleotides; the insertion of one or several nucleotides, such as by unequal recombination or gene conversion, resulting in an interruption of the coding sequence of the gene; duplication of all or a part of the gene; transposition of all or a part of the gene; or rearrangement of all or a part of the gene. More than one such mutation may be present in a single gene. Such sequence changes cause a mutation in the polypeptide encoded by a GPCR gene. For example, if the mutation is a frame shift mutation, the frame shift can result in a change in the encoded amino acids, and/or can result in the generation of a premature stop codon, causing generation of a truncated polypeptide. Alternatively, a polymoφhism associated with a susceptibility to a disease or condition associated with a GPCR can be a synonymous mutation in one or more nucleotides (i.e., a mutation that does not result in a change in the polypeptide encoded by a GPCR gene). Such a polymoφhism may alter splicing sites, affect the stability or transport of mRNA, or otherwise affect the transcription or translation of the gene. A GPCR gene that has any of the mutations described above is referred to herein as a "mutant gene."

In a first method of diagnosing a susceptibility to a susceptibility to a disease or condition associated with a GPCR, hybridization methods, such as Southern analysis, Northern analysis, or in situ hybridizations, can be used (see Current

Protocols in Molecular Biology, Ausubel, F. et al, Eds., John Wiley & Sons, including all supplements through 1999). For example, a biological sample from a test subject (a "test sample") of genomic DNA, RNA, or cDNA, is obtained from an individual suspected of having, being susceptible to or predisposed for, or carrying a defect for, a susceptibility to a disease or condition associated with a GPCR (the

"test individual"). The individual can be an adult, child, or fetus. The test sample can be from any source which contains genomic DNA, such as a blood sample, sample of amniotic fluid, sample of cerebrospinal fluid, or tissue sample from skin, muscle, buccal or conjunctival mucosa, placenta, gastrointestinal tract or other organs. A test sample of DNA from fetal cells or tissue can be obtained by appropriate methods, such as by amniocentesis or chorionic villus sampling. The DNA, RNA, or cDNA sample is then examined to determine whether a polymoφhism in a GPCR is present, and/or to determine which splicing variant(s) encoded by the GPCR is present. The presence of the polymoφhism or splicing variant(s) can be indicated by hybridization of the gene in the genomic DNA, RNA, or cDNA to a nucleic acid probe. A "nucleic acid probe", as used herein, can be a DNA probe or an RNA probe; the nucleic acid probe can contain at least one polymoφhism in a GPCR or contains a nucleic acid encoding a particular splicing variant of a GPCR. The probe can be any of the nucleic acid molecules described above (e.g., the gene, a fragment, a vector comprising the gene, a probe or primer, etc.).

To diagnose a susceptibility to a disease or condition associated with a GPCR, a test sample containing a GPCR, is contacted with at least one nucleic acid probe to form a hybridization sample. A preferred probe for detecting mRNA or genomic DNA is a labeled nucleic acid probe capable of hybridizing to mRNA or genomic

DNA sequences described herein. The nucleic acid probe can be, for example, a full-length nucleic acid molecule, or a portion thereof, such as an oligonucleotide of at least 15, 30, 50, 100, 250 or 500 nucleotides in length and sufficient to specifically hybridize under stringent conditions to appropriate mRNA or genomic DNA. For example, the nucleic acid probe can be all or a portion of one of SEQ ID

NOs: 1-124 (odd numbers), or the complement thereof, or a portion thereof; or can be a nucleic acid encoding a portion of one of SEQ ID NOs: 1-124 (even numbers). Other suitable probes for use in the diagnostic assays of the invention are described above (see e.g., probes and primers discussed under the heading, "Nucleic Acids of the Invention").

The hybridization sample is maintained under conditions that are sufficient to allow specific hybridization of the nucleic acid probe to a GPCR. "Specific hybridization", as used herein, indicates exact hybridization (e.g., with no mismatches). Specific hybridization can be performed under high stringency conditions or moderate stringency conditions, for example, as described above. In a particularly preferred embodiment, the hybridization conditions for specific hybridization are high stringency. Specific hybridization, if present, is then detected using standard methods. If specific hybridization occurs between the nucleic acid probe and the GPCR in the test sample, then the GPCR has the polymoφhism, or is the splicing variant, that is present in the nucleic acid probe. More than one nucleic acid probe can also be used concurrently in this method. Specific hybridization of any one of the nucleic acid probes is indicative of a polymoφhism in the GPCR, or of the presence of a particular splicing variant encoding the GPCR and is therefore diagnostic for a susceptibility to a susceptibility to a disease or condition associated with a GPCR.

In Northern analysis (see Current Protocols in Molecular Biology, Ausubel, F. et al, eds., John Wiley & Sons, supra) the hybridization methods described above are used to identify the presence of a polymoφhism or a particular splicing variant, associated with a susceptibility to a susceptibility to a disease or condition associated with a GPCR. For Northern analysis, a test sample of RNA is obtained from the individual by appropriate means. Specific hybridization of a nucleic acid probe, as described above, to RNA from the individual is indicative of a polymoφhism in a GPCR, or of the presence of a particular splicing variant encoded by a GPCR, and is therefore diagnostic for a susceptibility to a susceptibility to a disease or condition associated with a GPCR.

For representative examples of use of nucleic acid probes, see, for example, U.S. Patents No. 5,288,611 and 4,851,330. Alternatively, a peptide nucleic acid (PNA) probe can be used instead of a nucleic acid probe in the hybridization methods described above. PNA is a DNA mimic having a peptide-like, inorganic backbone, such as N-(2-aminoethyl)glycine units, with an organic base (A, G, C, T or U) attached to the glycine nitrogen via a methylene carbonyl linker (see, for example, Nielsen, P.E. et al, Bioconjugate Chemistry 5, American Chemical Society, p. 1 (1994). The PNA probe can be designed to specifically hybridize to a gene having a polymoφhism associated with a susceptibility to a susceptibility to a disease or condition associated with a GPCR. Hybridization of the PNA probe to a GPCR is diagnostic for a susceptibility to a susceptibility to a disease or condition associated with a GPCR.

In another method of the invention, mutation analysis by restriction digestion can be used to detect a mutant gene, or genes containing a polymoφhism(s), if the mutation or polymoφhism in the gene results in the creation or elimination of a restriction site. A test sample containing genomic DNA is obtained from the individual. Polymerase chain reaction (PCR) can be used to amplify a GPCR (and, if necessary, the flanking sequences) in the test sample of genomic DNA from the test individual. RFLP analysis is conducted as described (see Current Protocols in Molecular Biology, supra). The digestion pattern of the relevant DNA fragment indicates the presence or absence of the mutation or polymoφhism in the GPCR, and therefore indicates the presence or absence of this susceptibility to a susceptibility to a disease or condition associated with a GPCR.

Sequence analysis can also be used to detect specific polymoφhisms in a GPCR. A test sample of DNA or RNA is obtained from the test individual. PCR or other appropriate methods can be used to amplify the gene, and/or its flanking sequences, if desired. The sequence of a GPCR, or a fragment of the gene, or cDNA, or fragment of the cDNA, or mRNA, or fragment of the mRNA, is determined, using standard methods. The sequence of the gene, gene fragment, cDNA, cDNA fragment, mRNA, or mRNA fragment is compared with the known nucleic acid sequence of the gene, cDNA (e.g., one or more of SEQ ID NOs: 1-124 (odd numbers), or a complement thereof, or a nucleic acid sequence encoding one of SEQ ID NOs: 1-124 (even numbers) or a fragment thereof) or mRNA, as appropriate. The presence of a polymoφhism in the GPCR indicates that the individual has a susceptibility to a susceptibility to a disease or condition associated with a GPCR.

Allele-specific oligonucleotides can also be used to detect the presence of a polymoφhism in a GPCR, through the use of dot-blot hybridization of amplified oligonucleotides with allele-specific oligonucleotide (ASO) probes (see, for example, Saiki, R. et al, Nature 324:163-166 (1986)). An "allele-specific oligonucleotide" (also referred to herein as an "allele-specific oligonucleotide probe") is an oligonucleotide of approximately 10-50 base pairs, preferably approximately 15-30 base pairs, that specifically hybridizes to a GPCR, and that contains a polymoφhism associated with a susceptibility to a susceptibility to a disease or condition associated with a GPCR. An allele-specific oligonucleotide probe that is specific for particular polymoφhisms in a GPCR can be prepared, using standard methods (see Current Protocols in Molecular Biology, supra). To identify polymoφhisms in the gene that are associated with a susceptibility to a susceptibility to a disease or condition associated with a GPCR, a test sample of DNA is obtained from the individual. PCR can be used to amplify all or a fragment of a GPCR, and its flanking sequences. The DNA containing the amplified GPCR (or fragment of the gene) is dot-blotted, using standard methods (see Current Protocols in Molecular Biology, supra), and the blot is contacted with the oligonucleotide probe. The presence of specific hybridization of the probe to the amplified GPCR is then detected. Specific hybridization of an allele-specific oligonucleotide probe to DNA from the individual is indicative of a polymoφhism in the GPCR, and is therefore indicative of a susceptibility to a susceptibility to a disease or condition associated with a GPCR.

In another embodiment, arrays of oligonucleotide probes that are complementary to target nucleic acid sequence segments from an individual, can be used to identify polymoφhisms in a GPCR. For example, in one embodiment, an oligonucleotide array can be used. Oligonucleotide arrays typically comprise a plurality of different oligonucleotide probes that are coupled to a surface of a substrate in different known locations. These oligonucleotide arrays, also described as "Genechips™," have been generally described in the art, for example, U.S. Pat. No. 5,143,854 and PCT patent publication Nos. WO 90/15070 and 92/10092. These arrays can generally be produced using mechanical synthesis methods or light directed synthesis methods which incoφorate a combination of photolithographic methods and solid phase oligonucleotide synthesis methods. See Fodor et al, Science 251:767-777 (1991), Pirrung et al, U.S. Pat. No. 5,143,854 (see also PCT Application No. WO 90/15070) and Fodor et al, PCT Publication No. WO 92/10092 and U.S. Pat. No. 5,424,186, the entire teachings of each of which are incoφorated by reference herein. Techniques for the synthesis of these arrays using mechanical synthesis methods are described in, e.g., U.S. Pat. Nos. 5,384,261; the entire teachings of which are incoφorated by reference herein.

Once an oligonucleotide array is prepared, a nucleic acid of interest is hybridized with the array and scanned for polymoφhisms. Hybridization and scanning are generally carried out by methods described herein and also in, e.g. ,

Published PCT Application Nos. WO 92/10092 and WO 95/11995, and U.S. Pat. No. 5,424,186, the entire teachings of which are incoφorated by reference herein. In brief, a target nucleic acid sequence that includes one or more previously identified polymoφhic markers is amplified by well known amplification techniques, e.g., PCR. Typically, this involves the use of primer sequences that are complementary to the two strands of the target sequence both upstream and downstream from the polymoφhism. Asymmetric PCR techniques may also be used. Amplified target, generally incoφorating a label, is then hybridized with the array under appropriate conditions. Upon completion of hybridization and washing of the array, the array is scanned to determine the position on the array to which the target sequence hybridizes. The hybridization data obtained from the scan is typically in the form of fluorescence intensities as a function of location on the array.

Although primarily described in terms of a single detection block, e.g., for detection of a single polymoφhism, arrays can include multiple detection blocks, and thus be capable of analyzing multiple, specific polymoφhisms. In alternate arrangements, it will generally be understood that detection blocks may be grouped within a single array or in multiple, separate arrays so that varying, optimal conditions may be used during the hybridization of the target to the array. For example, it may often be desirable to provide for the detection of those polymoφhisms that fall within G-C rich stretches of a genomic sequence, separately from those falling in A-T rich segments. This allows for the separate optimization of hybridization conditions for each situation.

Additional description of use of oligonucleotide arrays for detection of polymoφhisms can be found, for example, in U.S. Patents 5,858,659 and 5,837,832, the entire teachings of which are incoφorated by reference herein.

Other methods of nucleic acid analysis can be used to detect polymoφhisms in a GPCR or variants encoding by a GPCR. Representative methods include direct manual sequencing (Church and Gilbert, Proc. Natl. Acad. Sci. USA 81:1991-1995 (1988); Sanger, F. et al. Proc. Natl. Acad. Sci. USA 74:5463-5467 (1977); Beavis et al, U.S. Pat. No. 5,288,644); automated fluorescent sequencing; single-stranded conformation polymoφhism assays (SSCP); clamped denaturing gel electrophoresis (CDGE); denaturing gradient gel electrophoresis (DGGE) (Sheffield, V.C. et al.

Proc. Natl. Acad. Sci. USA 86:232-236 (1989)), mobility shift analysis (Orita, M. et al, Proc. Natl. Acad. Sci. USA 86:2766-2770 (1989)), restriction enzyme analysis (Flavell et al, Cell 15:25 (1978); Geever, et al, Proc. Natl. Acad. Sci. USA 78:5081 (1981)); heteroduplex analysis; chemical mismatch cleavage (CMC) (Cotton et al, Proc. Natl. Acad. Sci. USA 85:4397-4401 (1985)); RNase protection assays (Myers,

RM. et al, Science 230:1242 (1985)); use of polypeptides which recognize nucleotide mismatches, such as E. coli mutS protein; allele-specific PCR, for example.

In another embodiment of the invention, diagnosis of a susceptibility to a susceptibility to a disease or condition associated with a GPCR can also be made by examining expression and/or composition of a GPCR polypeptide, by a variety of methods, including enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations and immunofluorescence. A test sample from an individual is assessed for the presence of an alteration in the expression and/or an alteration in composition of the polypeptide encoded by a GPCR, or for the presence of a particular variant encoded by a GPCR. An alteration in expression of a polypeptide encoded by a GPCR can be, for example, an alteration in the quantitative polypeptide expression (i.e., the amount of polypeptide produced); an alteration in the composition of a polypeptide encoded by a GPCR is an alteration in the qualitative polypeptide expression (e.g., expression of a mutant GPCR polypeptide or of a different splicing variant). In a preferred embodiment, diagnosis of a susceptibility to a disease or condition associated with a GPCR is made by detecting a particular splicing variant encoded by that GPCR, or a particular pattern of splicing variants.

Both such alterations (quantitative and qualitative) can also be present. An "alteration" in the polypeptide expression or composition, as used herein, refers to an alteration in expression or composition in a test sample, as compared with the expression or composition of polypeptide by a GPCR in a control sample. A control sample is a sample that corresponds to the test sample (e.g., is from the same type of cells), and is from an individual who is not affected by a susceptibility to a disease or condition associated with a GPCR. An alteration in the expression or composition of the polypeptide in the test sample, as compared with the control sample, is indicative of a susceptibility to a susceptibility to a disease or condition associated with a GPCR. Similarly, the presence of one or more different splicing variants in the test sample, or the presence of significantly different amounts of different splicing variants in the test sample, as compared with the control sample, is indicative of a susceptibility to a susceptibility to a disease or condition associated with a GPCR. Various means of examining expression or composition of the polypeptide encoded by a GPCR can be used, including speetroscopy, colorimetry, electrophoresis, isoelectric focusing, and immunoassays (e.g., David et al, U.S. Pat. No. 4,376,110) such as immunoblotting (see also Current Protocols in Molecular Biology, particularly Chapter 10). For example, in one embodiment, an antibody capable of binding to the polypeptide (e.g., as described above), preferably an antibody with a detectable label, can be used. Antibodies can be polyclonal, or more preferably, monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab')₂) can be used. The term "labeled", with regard to the probe or antibody, is intended to encompass direct labeling of the probe or antibody by coupling (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling of the probe or antibody by reactivity with another reagent that is directly labeled. Examples of indirect labeling include detection of a primary antibody using a fluorescently labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently labeled streptavidin.

Western blotting analysis, using an antibody as described above that specifically binds to a polypeptide encoded by a mutant GPCR, or an antibody that specifically binds to a polypeptide encoded by a non-mutant gene, or an antibody that specifically binds to a particular splicing variant encoded by a GPCR, can be used to identify the presence in a test sample of a particular splicing variant or of a polypeptide encoded by a polymoφhic or mutant GPCR, or the absence in a test sample of a particular splicing variant or of a polypeptide encoded by a non- polymoφhic or non-mutant gene. The presence of a polypeptide encoded by a polymoφhic or mutant gene, or the absence of a polypeptide encoded by a non- polymoφhic or non-mutant gene, is diagnostic for a susceptibility to a susceptibility to a disease or condition associated with a GPCR, as is the presence (or absence) of particular splicing variants encoded by the GPCR gene. In one embodiment of this method, the level or amount of polypeptide encoded by a GPCR in a test sample is compared with the level or amount of the polypeptide encoded by the GPCR in a control sample. A level or amount of the polypeptide in the test sample that is higher or lower than the level or amount of the polypeptide in the control sample, such that the difference is statistically significant, is indicative of an alteration in the expression of the polypeptide encoded by the

GPCR, and is diagnostic for a susceptibility to a susceptibility to a disease or condition associated with that GPCR. Alternatively, the composition of the polypeptide encoded by a GPCR in a test sample is compared with the composition of the polypeptide encoded by the GPCR in a control sample (e.g., the presence of different splicing variants). A difference in the composition of the polypeptide in the test sample, as compared with the composition of the polypeptide in the control sample, is diagnostic for a susceptibility to a susceptibility to a disease or condition associated with that GPCR. In another embodiment, both the level or amount and the composition of the polypeptide can be assessed in the test sample and in the control sample. A difference in the amount or level of the polypeptide in the test sample, compared to the control sample; a difference in composition in the test sample, compared to the control sample; or both a difference in the amount or level, and a difference in the composition, is indicative of a susceptibility to a susceptibility to a disease or condition associated with that GPCR. Kits (e.g. , reagent kits) useful in the methods of diagnosis comprise components useful in any of the methods described herein, including for example, hybridization probes or primers as decribed herein (e.g., labeled probes or primers), reagents for detection of labeled molecules, restriction enzymes (e.g., for RFLP analysis), allele-specific oligonucleotides, antibodies which bind to mutant or to non-mutant (native) GPCR polypeptide, means for amplification of nucleic acids comprising a GPCR, or means for analyzing the nucleic acid sequence of a GPCR or for analyzing the amino acid sequence of a GPCR polypeptide, etc.

SCREENING ASSAYS AND AGENTS IDENTIFIED THEREBY

The invention provides methods (also referred to herein as "screening assays") for identifying the presence of a nucleotide that hybridizes to a nucleic acid of the invention, as well as for identifying the presence of a polypeptide encoded by a nucleic acid of the invention. In one embodiment, the presence (or absence) of a nucleic acid molecule of interest (e.g., a nucleic acid that has significant homology with a nucleic acid of the invention) in a sample can be assessed by contacting the sample with a nucleic acid comprising a nucleic acid of the invention (e.g., a nucleic acid having the sequence of one of SEQ ID NOs: 1-124 (odd numbers), or the complement thereof, or a nucleic acid encoding an amino acid having the sequence of one of SEQ ID NOs:l-124 (even numbers), or a fragment or variant of such nucleic acids), under stringent conditions as described above, and then assessing the sample for the presence (or absence) of hybridization. In a preferred embodiment, high stringency conditions are conditions appropriate for selective hybridization. In another embodiment, a sample containing the nucleic acid molecule of interest is contacted with a nucleic acid containing a contiguous nucleotide sequence (e.g., a primer or a probe as described above) that is at least partially complementary to a part of the nucleic acid molecule of interest (e.g., a GPCR nucleic acid), and the contacted sample is assessed for the presence or absence of hybridization. In a preferred embodiment, the nucleic acid containing a contiguous nucleotide sequence is completely complementary to a part of the nucleic acid molecule of interest.

In any of these embodiments, all or a portion of the nucleic acid of interest can be subjected to amplification prior to performing the hybridization.

In another embodiment, the presence (or absence) of a polypeptide of interest, such as a polypeptide of the invention or a fragment or variant thereof, in a sample can be assessed by contacting the sample with an antibody that specifically hybridizes to the polypeptide of interest (e.g., an antibody such as those described above), and then assessing the sample for the presence (or absence) of binding of the antibody to the polypeptide of interest. In another embodiment, the invention provides methods for identifying agents

(e.g., fusion proteins, polypeptides, peptidomimetics, prodrugs, other receptors associated with GPCRs, binding agents, antibodies, small molecules or other drugs, or ribozymes which alter (e.g., increase or decrease) the activity of the polypeptides described herein, or which otherwise interact with the polypeptides herein. For example, such agents can be agents which bind to polypeptides described herein (e.g., GPCR binding agents); which have a stimulatory or inhibitory effect on, for example, activity of polypeptides of the invention; or which change (e.g., enhance or inhibit) the ability of the polypeptides of the invention to interact with GPCR binding agents (e.g, G-proteins, other receptors associated with GPCRs, or other binding agents); or which alter posttranslational processing of the GPCR polypeptide (e.g., agents that alter proteolytic processing to direct the polypeptide from where it is normally synthesized to another location in the cell, such as the cell surface; agents that alter proteolytic processing such that more polypeptide is released from the cell, etc.

In one embodiment, the invention provides assays for screening candidate or test agents that bind to or modulate the activity of polypeptides described herein (or biologically active portion(s) thereof), as well as agents identifiable by the assays. Test agents can be obtained using any of the numerous approaches in combinatorial library methods known in the art, including: biological libraries; spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the 'one-bead one-compound' library method; and synthetic library methods using affinity chromatography selection. The biological library approach is limited to polypeptide libraries, while the other four approaches are applicable to polypeptide, non-peptide oligomer or small molecule libraries of compounds (Lam, K.S. , Anticancer Drug Des. 12: 145 (1997)).

In one embodiment, to identify agents which alter the activity of a GPCR polypeptide, a cell, cell lysate, or solution containing or expressing a GPCR polypeptide (e.g., one of SEQ ID NOs:l-124 (even numbers), or another splicing variant encoded by a GPCR), or a fragment or derivative thereof (as described above), can be contacted with an agent to be tested; alternatively, the polypeptide can be contacted directly with the agent to be tested. The level (amount) of GPCR activity is assessed (e.g., the level (amount) of GPCR activity is measured, either directly or indirectly), and is compared with the level of activity in a control (i.e., the level of activity of the GPCR polypeptide or active fragment or derivative thereof in the absence of the agent to be tested). If the level of the activity in the presence of the agent differs, by an amount that is statistically significant, from the level of the activity in the absence of the agent, then the agent is an agent that alters the activity of a GPCR polypeptide. An increase in the level of GPCR activity relative to a control, indicates that the agent is an agent that enhances (is an agonist of) GPCR activity. Similarly, a decrease in the level of GPCR activity relative to a control, indicates that the agent is an agent that inhibits (is an antagonist of) GPCR activity. In another embodiment, the level of activity of a GPCR polypeptide or derivative or fragment thereof in the presence of the agent to be tested, is compared with a control level that has previously been established. A level of the activity in the presence of the agent that differs from the control level by an amount that is statistically significant indicates that the agent alters GPCR activity.

The present invention also relates to an assay for identifying agents which alter the expression of a GPCR gene (e.g., antisense nucleic acids, fusion proteins, polypeptides, peptidomimetics, prodrugs, other receptors associated with GPCRs, G- proteins, binding agents, antibodies, small molecules or other drugs, or ribozymes) which alter (e.g. , increase or decrease) expression (e.g. , transcription or translation) of the gene or which otherwise interact with the nucleic acids described herein, as well as agents identifiable by the assays. For example, a solution containing a nucleic acid encoding a GPCR polypeptide (e.g, a GPCR gene) can be contacted with an agent to be tested. The solution can comprise, for example, cells containing the nucleic acid or cell lysate containing the nucleic acid; alternatively, the solution can be another solution mat comprises elements necessary for transcription/translation of the nucleic acid. Cells not suspended in solution can also be employed, if desired. The level and/or pattern of GPCR expression (e.g., the level and/or pattern of mRNA or of protein expressed, such as the level and/or pattern of different splicing variants) is assessed, and is compared with the level and/or pattern of expression in a control (i.e., the level and/or pattern of the GPCR expression in the absence of the agent to be tested). If the level and/or pattern in the presence of the agent differ(s), by an amount or in a manner that is statistically significant, from the level and/or pattern in the absence of the agent, then the agent is an agent that alters the expression of GPCR. Enhancement of GPCR expression indicates that the agent is an agonist of GPCR activity. Similarly, inhibition of GPCR expression indicates that the agent is an antagonist of GPCR activity. In another embodiment, the level and/or pattern of GPCR polypeptide(s) (e.g., different splicing variants) in the presence of the agent to be tested, is compared with a control level and/or pattern that have previously been established. A level and/or pattern in the presence of the agent that differs from the control level and/or pattern by an amount or in a manner that is statistically significant indicates that the agent alters GPCR expression.

In another embodiment of the invention, agents which alter the expression of a GPCR gene or which otherwise interact with the nucleic acids described herein, can be identified using a cell, cell lysate, or solution containing a nucleic acid encoding the promoter region of the GPCR gene operably linked to a reporter gene. After contact with an agent to be tested, the level of expression of the reporter gene (e.g., the level of mRNA or of protein expressed) is assessed, and is compared with the level of expression in a control (z^'.e., the level of the expression of the reporter gene in the absence of the agent to be tested). If the level in the presence of the agent differs, by an amount or in a manner that is statistically significant, from the level in the absence of the agent, then the agent is an agent that alters the expression of the

GPCR, as indicated by its ability to alter expression of a gene that is operably linked to the GPCR gene promoter. Enhancement of the expression of the reporter indicates that the agent is an agonist of GPCR activity. Similarly, inhibition of the expression of the reporter indicates that the agent is an antagonist of GPCR activity. In another embodiment, the level of expression of the reporter in the presence of the agent to be tested, is compared with a control level that has previously been established. A level in the presence of the agent that differs from the control level by an amount or in a manner that is statistically significant indicates that the agent alters expression. Agents which alter the amounts of different splicing variants encoded by a

GPCR (e.g., an agent which enhances activity of a first splicing variant, and which inhibits activity of a second splicing variant), as well as agents which are agonists of activity of a first splicing variant and antagonists of activity of a second splicing variant, can easily be identified using these methods described above. In other embodiments of the invention, assays can be used to assess the impact of a test agent on the activity of a polypeptide in relation to a GPCR binding agent. For example, a cell that expresses a compound that interacts with a GPCR (herein referred to as a "GPCR binding agent", which can be a polypeptide or other molecule that interacts with a GPCR, such as a G-protein) is contacted with a GPCR in the presence of a test agent, and the ability of the test agent to alter the interaction between the GPCR and the GPCR binding agent is determined. Alternatively, a cell lysate or a solution containing the GPCR binding agent, can be used. An agent which binds to the GPCR or the GPCR binding agent can alter the interaction by interfering with, or enhancing the ability of the GPCR to bind to, associate with, or otherwise interact with the GPCR binding agent. Determining the ability of the test agent to bind to a GPCR or a GPCR binding agent can be accomplished, for example, by coupling the test agent with a radioisotope or enzymatic label such that binding of the test agent to the polypeptide can be determined by detecting the

125 35 14 3 labeled with I, S, C or H, either directly or indirectly, and the radioisotope detected by direct counting of radioemmission or by scintillation counting.

Alternatively, test agents can be enzymatically labeled with, for example, horseradish peroxidase, alkaline phosphatase, or luciferase, and the enzymatic label detected by determination of conversion of an appropriate substrate to product. It is also within the scope of this invention to determine the ability of a test agent to interact with the polypeptide without the labeling of any of the interactants. For example, a microphysiometer can be used to detect the interaction of a test agent with a GPCR or a GPCR binding agent without the labeling of either the test agent, GPCR, or the GPCR binding agent. McConnell, H.M. et al. , Science 257:1906-

1912 (1992). As used herein, a "microphysiometer" (e.g., Cytosensor™) is an analytical instrument that measures the rate at which a cell acidifies its environment using a light-addressable potentiometric sensor (LAPS). Changes in this acidification rate can be used as an indicator of the interaction between ligand and polypeptide. Thus, these receptors can be used to screen for compounds that are agonists for use in treating a susceptibility to a disease or condition associated with a GPCR or antagonists for studying a susceptibility to a disease or condition associated with a GPCR. Drugs could be designed to regulate GPCR activation that in turn can be used to regulate signaling pathways and transcription events of genes downstream.

In another embodiment of the invention, assays can be used to identify polypeptides that interact with one or more GPCR polypeptides, as described herein. For example, a yeast two-hybrid system such as that described by Fields and Song (Fields, S. and Song, O., Nature 340:245-246 (1989)) can be used to identify polypeptides that interact with one or more GPCR polypeptides. In such a yeast two-hybrid system, vectors are constructed based on the flexibility of a transcription factor that has two functional domains (a DNA binding domain and a transcription activation domain). If the two domains are separated but fused to two different proteins that interact with one another, transcriptional activation can be achieved, and transcription of specific markers (e.g, nutritional markers such as His and Ade, or color markers such as lacZ) can be used to identify the presence of interaction and transcriptional activation. For example, in the methods of the invention, a first vector is used which includes a nucleic acid encoding a DNA binding domain and also a GPCR polypeptide, splicing variant, or fragment or derivative thereof, and a second vector is used which includes a nucleic acid encoding a transcription activation domain and also a nucleic acid encoding a polypeptide which potentially may interact with the GPCR polypeptide, splicing variant, or fragment or derivative thereof (e.g., a GPCR polypeptide binding agent or G-protein). Incubation of yeast containing the first vector and the second vector under appropriate conditions (e.g., mating conditions such as used in the Matchmaker™ system from Clontech (Palo

Alto, California, USA)) allows identification of colonies that express the markers of interest. These colonies can be examined to identify the polypeptide(s) that interact with the GPCR polypeptide or fragment or derivative thereof. Such polypeptides may be useful as agents that alter the activity of expression of a GPCR polypeptide, as described above.

In more than one embodiment of the above assay methods of the present invention, it may be desirable to immobilize either GPCR, the GPCR binding agent, or other components of the assay on a solid support, in order to facilitate separation of complexed from uncomplexed forms of one or both of the polypeptides, as well as to accommodate automation of the assay. Binding of a test agent to the polypeptide, or interaction of the polypeptide with a binding agent in the presence and absence of a test agent, can be accomplished in any vessel suitable for containing the reactants. Examples of such vessels include microtiter plates, test tubes, and micro-centrifuge tubes. In one embodiment, a fusion protein (e.g., a glutathione-S-transferase fusion protein) can be provided which adds a domain that allows GPCR or a GPCR binding agent to be bound to a matrix or other solid support. In another embodiment, modulators of expression of nucleic acid molecules of the invention are identified in a method wherein a cell, cell lysate, or solution containing a nucleic acid encoding a GPCR is contacted with a test agent and the expression of appropriate mRNA or polypeptide (e.g., splicing variant(s)) in the cell, cell lysate, or solution, is determined. The level of expression of appropriate mRNA or polypeptide(s) in the presence of the test agent is compared to the level of expression of mRNA or polypeptide(s) in the absence of the test agent. The test agent can then be identified as a modulator of expression based on this comparison. For example, when expression of mRNA or polypeptide is greater (statistically significantly greater) in the presence of the test agent than in its absence, the test agent is identified as a stimulator or enhancer of the mRNA or polypeptide expression. Alternatively, when expression of the mRNA or polypeptide is less (statistically significantly less) in the presence of the test agent than in its absence, the test agent is identified as an inhibitor of the mRNA or polypeptide expression. The level of mRNA or polypeptide expression in the cells can be determined by methods described herein for detecting mRNA or polypeptide.

This invention further pertains to novel agents identified by the above- described screening assays. Accordingly, it is within the scope of this invention to further use an agent identified as described herein in an appropriate animal model. For example, an agent identified as described herein (e.g., a test agent that is a modulating agent, an antisense nucleic acid molecule, a specific antibody, or a polypeptide-binding agent) can be used in an animal model to determine the efficacy, toxicity, or side effects of treatment with such an agent. Alternatively, an agent identified as described herein can be used in an animal model to determine the mechanism of action of such an agent. Furthermore, this invention pertains to uses of novel agents identified by the above-described screening assays for treatments as described herein. In addition, an agent identified as described herein can be used to alter activity of a polypeptide encoded by a GPCR, or to alter expression of a GPCR, by contacting the polypeptide or the gene (or contacting a cell comprising the polypeptide or the gene) with the agent identified as described herein.

PHARMACEUTICAL COMPOSITIONS

The present invention also pertains to pharmaceutical compositions comprising nucleic acids described herein, particularly nucleotides encoding the polypeptides described herein; comprising polypeptides described herein (e.g., one or more of SEQ ID NOs:l-124 (even numbers)); and/or comprising other splicing variants encoded by a GPCR; and/or an agent that alters (e.g., enhances or inhibits) GPCR gene expression or GPCR polypeptide activity as described herein. For instance, a polypeptide, protein (e.g., a G-protein), an agent that alters GPCR gene expression, or a GPCR binding agent or binding partner, fragment, fusion protein or prodrug thereof, or a nucleotide or nucleic acid construct (vector) comprising a nucleotide of the present invention, or an agent that alters GPCR polypeptide activity, can be formulated with a physiologically acceptable carrier or excipient to prepare a pharmaceutical composition. The carrier and composition can be sterile.

The formulation should suit the mode of admimstration.

Suitable pharmaceutically acceptable carriers include but are not limited to water, salt solutions (e.g., NaCl), saline, buffered saline, alcohols, glycerol, ethanol, gum arabic, vegetable oils, benzyl alcohols, polyethylene glycols, gelatin, carbohydrates such as lactose, amylose or starch, dextrose, magnesium stearate, talc, silicic acid, viscous paraffin, perfume oil, fatty acid esters, hydroxymethylcellulose, polyvinyl pyrolidone, etc., as well as combinations thereof. The pharmaceutical preparations can, if desired, be mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, flavoring and/or aromatic substances and the like which do not deleteriously react with the active agents.

The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. The composition can be a liquid solution, suspension, emulsion, tablet, pill, capsule, sustained release formulation, or powder. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulation can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, polyvinyl pyrollidone, sodium saccharine, cellulose, magnesium carbonate, etc.

Methods of introduction of these compositions include, but are not limited to, intradermal, intramuscular, intraperitoneal, intraocular, intravenous, subcutaneous, topical, oral and intranasal. Other suitable methods of introduction can also include gene therapy (as described below), rechargeable or biodegradable devices, particle acceleration devises ("gene guns") and slow release polymeric devices. The pharmaceutical compositions of this invention can also be administered as part of a combinatorial therapy with other agents. The composition can be formulated in accordance with the routine procedures as a pharmaceutical composition adapted for administration to human beings. For example, compositions for intravenous administration typically are solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a solubilizing agent and a local anesthetic to ease pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampule or sachette indicating the quantity of active agent. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water, saline or dextrose/water. Where the composition is administered by injection, an ampule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to admimstration.

For topical application, nonsprayable forms, viscous to semi-solid or solid forms comprising a carrier compatible with topical application and having a dynamic viscosity preferably greater than water, can be employed. Suitable formulations include but are not limited to solutions, suspensions, emulsions, creams, ointments, powders, enemas, lotions, sols, liniments, salves, aerosols, etc., which are, if desired, sterilized or mixed with auxiliary agents, e.g., preservatives, stabilizers, wetting agents, buffers or salts for influencing osmotic pressure, etc. The agent may be incoφorated into a cosmetic formulation. For topical application, also suitable are sprayable aerosol preparations wherein the active ingredient, preferably in combination with a solid or liquid inert carrier material, is packaged in a squeeze bottle or in admixture with a pressurized volatile, normally gaseous propellant, e.g., pressurized air. Agents described herein can be formulated as neutral or salt forms.

Pharmaceutically acceptable salts include those formed with free amino groups such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with free carboxyl groups such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.

The agents are administered in a therapeutically effective amount. The amount of agents which will be therapeutically effective in the treatment of a particular disorder or condition will depend on the nature of the disorder or condition, and can be determined by standard clinical techniques. In addition, in vitro or in vivo assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the seriousness of the symptoms of a susceptibility to a disease or condition associated with a GPCR, and should be decided according to the judgment of a practitioner and each patient's circumstances. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

The invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention. Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use of sale for human administration. The pack or kit can be labeled with information regarding mode of administration, sequence of drug administration (e.g., separately, sequentially or concurrently), or the like. The pack or kit may also include means for reminding the patient to take the therapy. The pack or kit can be a single unit dosage of the combination therapy or it can be a plurality of unit dosages. In particular, the agents can be separated, mixed together in any combination, present in a single vial or tablet. Agents assembled in a blister pack or other dispensing means is preferred. For the puφose of this invention, unit dosage is intended to mean a dosage that is dependent on the individual pharmacodynamics of each agent and administered in FDA approved dosages in standard time courses.

METHODS OF THERAPY

The present invention also pertains to methods of treatment (prophylactic and or therapeutic) for a susceptibility to a disease or condition associated with a GPCR, using a GPCR therapeutic agent. A "GPCR therapeutic agent" is an agent that alters (e.g., enhances or inhibits) GPCR polypeptide activity and/or GPCR gene expression, as described herein (e.g., a GPCR agonist or antagonist). GPCR therapeutic agents can alter GPCR polypeptide activity or gene expression by a variety of means, such as, for example, by providing additional GPCR polypeptide or by upregulating the transcription or translation of the GPCR gene; by altering posttranslational processing of the GPCR polypeptide; by altering transcription of GPCR splicing variants; or by interfering with GPCR polypeptide activity (e.g., by binding to a GPCR polypeptide), or by downregulating the transcription or translation of a GPCR gene. Representative GPCR therapeutic agents include the following: nucleic acids or fragments or derivatives thereof described herein, particularly nucleotides encoding the polypeptides described herein and vectors comprising such nucleic acids (e.g., a gene, cDNA, and/or mRNA, such as a nucleic acid encoding a GPCR polypeptide or active fragment or derivative thereof, or an oligonucleotide; for example, one of SEQ ID NOs: 1-124 (odd numbers), or a complement thereof, or a nucleic acid encoding one of SEQ ID NOs: 1-124 (even numbers), or fragments or derivatives thereof); polypeptides described herein (e.g, one or more of SEQ ID NOs: 1-124 (even numbers), and/or other splicing variants encoded by a GPCR, or fragments or derivatives thereof); other polypeptides (e.g., G-proteins); GPCR binding agents; peptidomimetics; fusion proteins or prodrugs thereof; antibodies (e.g., an antibody to a mutant GPCR polypeptide, or an antibody to a non-mutant GPCR polypeptide, or an antibody to a particular splicing variant encoded by a GPCR, as described above); ribozymes; other small molecules; and other agents that alter (e.g, enhance or inhibit) GPCR gene expression or polypeptide activity, or that regulate transcription of GPCR splicing variants (e.g., agents that affect which splicing variants are expressed, or that affect the amount of each splicing variant that is expressed.

More than one GPCR therapeutic agent can be used concurrently, if desired. A GPCR therapeutic agent that is a nucleic acid is used in the treatment of a susceptibility to a disease or condition associated with a GPCR. The term, "treatment" as used herein, refers not only to ameliorating symptoms associated with the disease, but also preventing or delaying the onset of the disease, and also lessening the severity or frequency of symptoms of the disease. The therapy is designed to alter (e.g., inhibit or enhance), replace or supplement activity of a GPCR polypeptide in an individual. For example, a GPCR therapeutic agent can be administered in order to upregulate or increase the expression or availability of the GPCR gene or of specific splicing variants of GPCR, or, conversely, to downregulate or decrease the expression or availability of the GPCR gene or specific splicing variants of the GPCR. Upregulation or increasing expression or availability of a native GPCR gene or of a particular splicing variant could interfere with or compensate for the expression or activity of a defective gene or another splicing variant; downregulation or decreasing expression or availability of a native GPCR gene or of a particular splicing variant could minimize the expression or activity of a defective gene or the particular splicing variant and thereby minimize the impact of the defective gene or the particular splicing variant.

The GPCR therapeutic agent(s) are administered in a therapeutically effective amount (i.e., an amount that is sufficient to treat the disease, such as by ameliorating symptoms associated with the disease, preventing or delaying the onset of the disease, and or also lessening the severity or frequency of symptoms of the disease). The amount which will be therapeutically effective in the treatment of a particular individual's disorder or condition will depend on the symptoms and severity of the disease, and can be determined by standard clinical techniques. In addition, in vitro or in vivo assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the seriousness of the disease or disorder, and should be decided according to the judgment of a practitioner and each patient's circumstances. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

In one embodiment, a nucleic acid of the invention (e.g., a nucleic acid encoding a GPCR polypeptide, such as one of SEQ ID NOs: 1-124 (odd numbers), or a complement thereof; or another nucleic acid that encodes a GPCR polypeptide or a splicing variant, derivative or fragment thereof, such as a nucleic acid encoding one of SEQ ID NOs: 1-124 (even numbers)) can be used, either alone or in a pharmaceutical composition as described above. For example, a GPCR or a cDNA encoding a GPCR polypeptide, either by itself or included within a vector, can be introduced into cells (either in vitro or in vivo) such that the cells produce native GPCR polypeptide. If necessary, cells that have been transformed with the gene or cDNA or a vector comprising the gene or cDNA can be introduced (or re- introduced) into an individual affected with the disease. Thus, cells which, in nature, lack native GPCR expression and activity, or have mutant GPCR expression and activity, or have expression of a disease-associated GPCR splicing variant, can be engineered to express the GPCR polypeptide or an active fragment of the GPCR polypeptide (or a different variant of the GPCR polypeptide). In a preferred embodiment, nucleic acid encoding a GPCR polypeptide, or an active fragment or derivative thereof, can be introduced into an expression vector, such as a viral vector, and the vector can be introduced into appropriate cells in an animal. Other gene transfer systems, including viral and nonviral transfer systems, can be used. Alternatively, nonviral gene transfer methods, such as calcium phosphate coprecipitation, mechanical techniques (e.g., microinjection); membrane fusion- mediated transfer via liposomes; or direct DNA uptake, can also be used.

Alternatively, in another embodiment of the invention, a nucleic acid of the invention; a nucleic acid complementary to a nucleic acid of the invention; or a portion of such a nucleic acid (e.g., an oligonucleotide as described below), can be used in "antisense" therapy, in which a nucleic acid (e.g., an oligonucleotide) which specifically hybridizes to the mRNA and or genomic DNA of a GPCR is administered or generated in situ. The antisense nucleic acid that specifically hybridizes to the mRNA and/or DNA inhibits expression of the GPCR polypeptide, e.g., by inhibiting translation and/or transcription. Binding of the antisense nucleic acid can be by conventional base pair complementarity, or, for example, in the case of binding to DNA duplexes, through specific interaction in the major groove of the double helix.

An antisense construct of the present invention can be delivered, for example, as an expression plasmid as described above. When the plasmid is transcribed in the cell, it produces RNA that is complementary to a portion of the mRNA and/or DNA which encodes the GPCR polypeptide. Alternatively, the antisense construct can be an oligonucleotide probe that is generated ex vivo and introduced into cells; it then inhibits expression by hybridizing with the mRNA and/or genomic DNA of the GPCR. In one embodiment, the oligonucleotide probes are modified oligonucleotides that are resistant to endogenous nucleases, e.g. exonucleases and/or endonucleases, thereby rendering them stable in vivo. Exemplary nucleic acid molecules for use as antisense oligonucleotides are phosphoramidate, phosphothioate and methylphosphonate analogs of DNA (see also U.S. 5,176,996; 5,264,564; and 5,256,775). Additionally, general approaches to constructing oligomers useful in antisense therapy are also described, for example, by Van der Krol et al, (Biotechniques 6:958-976 (1988)); and Stein et al. (Cancer Res. 48:2659-2668 (1988)). With respect to antisense DNA, oligodeoxyribonucleotides derived from the translation initiation site are preferred. To perform antisense therapy, oligonucleotides (mRNA, cDNA or DNA) are designed that are complementary to mRNA encoding the GPCR. The antisense oligonucleotides bind to GPCR mRNA transcripts and prevent translation. Absolute complementarity, although preferred, is not required. A sequence "complementary" to a portion of an RNA, as refeπed to herein, indicates that a sequence has sufficient complementarity to be able to hybridize with the RNA, forming a stable duplex; in the case of double-stranded antisense nucleic acids, a single strand of the duplex DNA may thus be tested, or triplex formation may be assayed. The ability to hybridize will depend on both the degree of complementarity and the length of the antisense nucleic acid, as described in detail above. Generally, the longer the hybridizing nucleic acid, the more base mismatches with an RNA it may contain and still form a stable duplex (or triplex, as the case may be). One skilled in the art can ascertain a tolerable degree of mismatch by use of standard procedures. The oligonucleotides used in antisense therapy can be DNA, RNA, or chimeric mixtures or derivatives or modified versions thereof, single-stranded or double-stranded. The oligonucleotides can be modified at the base moiety, sugar moiety, or phosphate backbone, for example, to improve stability of the molecule, hybridization, etc. The oligonucleotides can include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger et al, Proc. Natl. Acad. Sci. USA 86:6553-6556 (1989); Lemaitre et al., Proc. Natl. Acad Sci. USA 84:648-652 (1987); PCT International Publication No. WO 88/09810) or the blood-brain barrier (see, e.g., PCT International Publication No. WO 89/10134), or hybridization- triggered cleavage agents (see, e.g., Krol et al, Bio/Techniques 6:958-976 (1988)) or intercalating agents. (See e.g., Zon, Pharm. Res. 5: 539-549 (1988)). To this end, the oligonucleotide may be conjugated to another molecule (e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent). The antisense molecules are delivered to cells that express GPCR in vivo. A number of methods can be used for delivering antisense DNA or RNA to cells; e.g., antisense molecules can be injected directly into the tissue site, or modified antisense molecules, designed to target the desired cells (e.g., antisense linked to peptides or antibodies that specifically bind receptors or antigens expressed on the target cell surface) can be administered systemically. Alternatively, in a preferred embodiment, a recombinant DNA construct is utilized in which the antisense oligonucleotide is placed under the control of a strong promoter (e.g., pol HI or pol TT). The use of such a construct to transfect target cells in the patient results in the transcription of sufficient amounts of single stranded RNAs that will form complementary base pairs with the endogenous GPCR transcripts and thereby prevent translation of the GPCR mRNA. For example, a vector can be introduced in vivo such that it is taken up in a cell and directs the transcription of an antisense RNA. Such a vector can remain episomal or become chromosomally integrated, as long as it can be transcribed to produce the desired antisense RNA. Such vectors can be constructed by recombinant DNA technology methods standard in the art and described above. For example, a plasmid, cosmid, YAC or viral vector can be used to prepare the recombinant DNA construct that can be introduced directly into the tissue site. Alternatively, viral vectors can be used which selectively infect the desired tissue, in which case administration may be accomplished by another route (e.g., systemically).

Endogenous GPCR expression can be reduced by inactivating or "knocking out" GPCR or its promoter using targeted homologous recombination (e.g., see Smithies et al, Nature 317:230-234 (1985); Thomas & Capecchi, Cell 51:503-512

(1987); Thompson et al, Cell 5:313-321 (1989)). For example, a mutant, nonfunctional GPCR (or a completely unrelated DNA sequence) flanked by DNA homologous to the endogenous GPCR (either the coding regions or regulatory regions of GPCR) can be used, with or without a selectable marker and/or a negative selectable marker, to transfect cells that express the GPCR in vivo. Insertion of the

DNA construct, via targeted homologous recombination, results in inactivation of the GPCR. The recombinant DNA constructs can be directly administered or targeted to the required site in vivo using appropriate vectors, as described above. Alternatively, expression of non-mutant GPCRs can be increased using a similar method: targeted homologous recombination can be used to insert a DNA construct comprising a non-mutant, functional GPCR, e.g., a gene having one of SEQ ED NOs: 1-124 (odd numbers), or the complement thereof, or a portion thereof, in place of a mutant GPCR in the cell, as described above. In another embodiment, targeted homologous recombination can be used to insert a DNA construct comprising a nucleic acid that encodes a GPCR polypeptide variant that differs from that present in the cell.

Alternatively, endogenous GPCR expression can be reduced by targeting deoxyribonucleotide sequences complementary to the regulatory region of a GPCR (i.e., the GPCR promoter and/or enhancers) to form triple helical structures that prevent transcription of the GPCR in target cells in the body. (See generally, Helene,

C, Anticancer Drug Des. 6(6): 569-84 (1991); Helene, C, et al, Ann. NY. Acad. Sci. 660:27-36 (1992); and Maher, L. J., Bioassays 14(12):807-15 (1992)). Likewise, the antisense constructs described herein, by antagonizing the normal biological activity of one of the GPCR proteins, can be used in the manipulation of tissue, e.g., tissue differentiation, both in vivo and for ex vivo tissue cultures.

Furthermore, the anti-sense techniques (e.g., microinjection of antisense molecules, or transfection with plasmids whose transcripts are anti-sense with regard to a GPCR mRNA or gene sequence) can be used to investigate the role of one or GPCR in developmental events, as well as the normal cellular function of the GPCRs in adult tissue. Such techniques can be utilized in cell culture, but can also be used in the creation of transgenic animals.

In yet another embodiment of the invention, other GPCR therapeutic agents as described herein can also be used in the treatment or prevention of a susceptibility to a disease or condition associated with a GPCR. The therapeutic agents can be delivered in a composition, as described above, or by themselves. They can be administered systemically, or can be targeted to a particular tissue. The therapeutic agents can be produced by a variety of means, including chemical synthesis; recombinant production; in vivo production (e.g., a transgenic animal, such as U.S.

Pat. No. 4,873,316 to Meade et al), for example, and can be isolated using standard means such as those described herein.

A combination of any of the above methods of treatment (e.g., administration of non-mutant GPCR polypeptide in conjunction with antisense therapy targeting mutant GPCR mRNA; administration of a first splicing variant encoded by a GPCR in conjunction with antisense therapy targeting a second splicing encoded by a GPCR), can also be used.

The teachings of all publications cited herein are incoφorated herein by reference in their entirety.

While this invention has been particularly shown and described with references to prefeπed embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.

Table I

MOOSE00162 ctgl4797448003..448091, 561616..561669, 625017..625027,

718931..719797, 720713..720771

MIQTISNVSEAVVQIIASQMPDGDNTDFRYFIYAVTYTVILVPGLIGNILALWV FYGYMKETKPAVIFMINLAIADLLQVLSLPLPJFYYLNHDWPFGPGLCMFCFYLKYV NMYASIYFLVCISVRRFWFLMYPFRFHDCKQKYDLYISIAGWLIICLACVLFPLLRTS DDTSGNRTKCFVDLPTRNV-π.AQSVVMMTIGELIGFVTPLLrvXYCTWKTVLSLQDK YPMAQDLGEKQKALKMILTCAGVFLICFAPYHFSFPLDFLVKSNEIKSCLARRVILIFH SVALCLASLNSCLDPVIYYFSTNEFKKSFLEQFRVIASTQEDHSSAEERISELEDRLLEN IQSEETKEKRIKNNE (SEQ ED NO: 2) atgattcagacaatcagtaatgttagtgaagcagtagtccaaatcatagcttctcaaatgccagatggagacaatacagatttt cgatactttatttatgcagtgacatacactgtcattcttgtgccaggtctcatagggaatata^ agaaacaaaacgagctgtgatatttatgataaacttagccattgctgacttactacaagttctttccttgccactgaggatcttctactacttg aatcatgactggccatttgggcctggtctctgcatgttctgtttctacctgaagtatgtcaacatgtatgcaagcatctacttcttggtctgca tcagtgtgcgacgattttggtttctcatgtacccctttcgcttccatgactgcaaacagaaatatgacctgtacatcagcattgctggctggc tgatcatctgccttgcctgtgtactctttccactcctcagaaccagtgatgatacctctggcaataggaccaaatgctttgtggatcttccta ccaggaatgtcaacctggcccagtccgttgttatgatgaccattggcgagttgattgggtttgtaactccgcttctgattgtcctatattgta cctggaagacggttttatcactgcaagataaatatcccatggcccaagatcttggagagaaacagaaagccttgaagatgattctaacct gtgcagggg^ttcctaatttgctttgcaccttatcatttcagttttcctttagatttcctggtgaagtccaatgaaattaaaagctgcctagcc agaagggtgattctaatatttcattctgtggcattgtgtcttgctagtctgaattcatg^cttgacccagtcatalactacttttccactaatgaa tttaaaaagtccttcctggaacaattcagggtgattgcatccacacaagaggatcactcctcagcagaagaaagaattagtgagcttgaa gacaggctacttgaaaatatacagtcagaggagacaaaagaaaaaagaataaaaaacaatgaa (SEQ ED NO: 1)

MOOSE00638 ctgl6465 1516512..1517438, 1589966..1590028

KSVLLYTLSFIYIFIFVTGMIANSVVΥWVNIQAKTTGYDTHCYILNLAIADLWV VLTIPVWVVSLVQHNQWPMGELTCKVTHLIFSINLFGSIFFLTCMSVDRYLSITYFTN TPSSRKKMVRRVNCILVWLLAFCVSLPDTYYLKTVTSASNNETYCRSFYPEHSIKEW LIGMELVSVVLGFAVPFSΠA YFLLARAISASSDQEKHSSRKΠFSYVVVFLVCWLPY FΓV^'AVLLDIFSILHYIPFTCRLEFΪALFTALHVTQCLSLVHCCVNPVXYSFΓNRNYRYEL MKAFIFKYSAKTGLTKLRDASRAHTRMHTHTHAHMHTHIHSLS (SEQ ED NO: 4) aaaagcgtcctgctctacacgctctccttcatttacattttcatcttcgtcatcggcatgattgccaactccgtggtggtctgggt gaatatccaggccaagaccacaggctatgacacgcactgctacatcttgaacctggccattgccgacctgtgggttgtcctcaccatcc cagtctgggtggtcagtctcgtgcagcacaaccagtggcccatgggcgagctcacgtgcaaagtcacacacctcatcttctccatcaa cctcttcggcagcattttcttcctcacgtgcatgagcgtggaccgctacctctccatcacctacttcaccaacacccccagcagcaggaa gaagatggtacgccgtgtcgtctgcatcctggtgtggctgctggccttctgcgtgtctctgcctgacacctactacctgaagaccgtcac gtctgcgtccaacaatgagacctactgccggtccttctaccccgagcacagcatcaaggagtggctgatcggcatggagctggtctcc gttgtcttgggctttgccgttcccttctccattatcgctgtcttctacttcctgctggccagagccatctcggcgtccagtgaccaggagaa gcacagcagccggaagatcatcttctcctacgtggtggtcttccttgtctgctggctgccctaccacgtggcggtgctgctggacatctt ctccatcctgcactacatccctttcacctgccggctggagcacgccctcttcacggccctgcatgtcacacagtgcctgtcgctggtgca ctgctgcgtcaaccctgtcctctacagcttcatcaatcgcaactacaggtacgagctgatgaaggccttcatcttcaagtactcggccaa aacagggctcaccaagctcatcgatgcctccagagcacacacgcgcatgcacacacacacacatgcacacatgcacacacacatac attctctttct (SEQ ED NO: 3)

MOOSE00693 ctgl6008 12028107..12028186, 12152632..12153541

IPLKMHYLPVIYGIΠ^?LVGFPGNA\^STYIFKMRPWKSSTIIML_Π.ACTDLLYL TSLPFLIHYYASGENWIFGDFMCKTIRFSFHFNLYSSILFLTCFSIFRYCVITHPMSCFSIH KTRCAVVACAVVWΠSLVAVIPMTFLITSTNRTNRSACLDLTSSDELNΉKWYNLILT ATTFCLPLVIVTLCYTTIMTLTHGLQTDSCLKQKARRLTILLLLAFYVCFLPFHILRVI

RIESRLLSISCSffiNQfflEAYIVSRPLAALNTFGNLLLYV SDNFQQAVCSTVRCKVS GNLEQAKKIMVSSSSSSHY HHHHHHHHHLHPDSQKI (SEQ ED NO: 6) atcccactcaagatgcactacctccctgttatttatggcattatcttcctcgtgggatttccaggcaatgcagtagtgatatccac ttacattttcaaaatgagaccttggaagagcagcaccatcattatgctgaacctggcctgcacagatctgctgtatctgaccagcctcccc ttcctgattcactøclatgccagtggcgaaaactggatctttggagatttcatgtgtaagtttatccgcttcagcttccatttcaacctgtatag cagcatcctcttcctcacctgtttcagcatcttccgctactgtg^gatcattcacccaatgagctgcttttccattcacaaaactcgatgtgca gttgtagcctgtgctgtggtgtggatcatttcactggtagctgtcattccgatgaccttcttgatcacatcaaccaacaggaccaacagatc agcctgtctcgacctcaccagttcggatgaactcaatactattaagtggtacaacctgattttgactgcaactactttctgcctccccttggt gatagtgacactttgctataccacgattatccacactctgacccatggactgcaaactgacagctgccttaagcagaaagcacgaaggc taaccattctgctactccttgcattttacgtatgltttttacccttecatatett ttccattgagaatcagatccatgaagcttacatcgtttctagaccattagctgctctgaacacctttggtaacctgttactatatgtggtggtc agcgacaactttcagcaggctgtctgctcaacagtgagatgcaaagtaagcgggaaccttgagcaagcaaagaaaattatggtatcat cttcatcatcatcacattatcatcatcatcatcatcatcatcaccatttacatccagacagccagaaaata (SEQ ED NO: 5)

MOOSE00717 ctgl6282 864967..865847, 878276..878384

FLALRLMVALAYGLVGAIGLLGNLAVLWVLSNCARRAPGPPSDTFVFNLALA DLGLALTLPFWAAESALDFHWPFGGALCKMVLTATVLNVYASIFLITALSVARYWV VAMAAGPGTHLSLFWARIATLAVWAAAALVTVPTAVFGVEGEVCGVRLCLLRFPSR YWLGAYQLQRVVLAFMVPLGVITTSYLLLLAFLQRRQRRRQDSRVVARSVRILVAS FFLCWFPNFrVNTLWGVLVKFDLVPWNSTFYTIQTYVFPVTTCLAHSNSCLNPVLYCL LRREPRQALAGTFRDLRAASCLRASSASRSSSARACSLSRSRASRLPASVARS (SEQ ED NO: 8) ttcctagccctgaggctcatggttgccctggcctatgggcttgtgggggccattggcttgctgggaaatttggcggtgctgtg ggtactgagtaactgtgcccggagagcccctggcccaccttcagacaccttcgtcttcaacctggctctggcggacctgggactggca ctcactctccccttttgggcagccgagtcggcactggactttcactggcccttcggaggtgccctctgcaagatggttctgacggccact gtcctcaacgtctatgccagcatcttcctcatcacagcgctgagcgttgctcgctactgggtggtggccatggctgcggggccaggca cccacctctcactcttctgggcccgaatagccaccctggcagtgtgggcggcggctgccctggtgacggtgcccacagctgtcttcgg ggtggagggtgaggtgtgtggtgtgcgcctttgcctgctgcgtttccccagcaggtactggctgggggcctaccagctgcagagggt ggtgctggctttcatggtgcccttgggcgtcatcaccaccagctacctgctgctgctggccttcctgcagcggcggcaacggcggcgg caggacagcagggtcgtggcccgctctgtccgcatcctggtggcttccttcttcctctgctggtttcccaaccatgtggtcactctctggg gtgtcctggtgaagtttgacctggtgccctggaacagtactttctatactatccagacgtatgtcttccctgtcactacttgcttggcacaca gcaatagctgcctcaaccctgtgctgtactgtctcctgaggcgggagccccggcaggctctggcaggcaccttcagggatctgcggg cggccagctgccttcgagcctcttcggcctcacgctccagcagtgcccgggcctgctcactctcccggagccgggcctccaggctgc cagcttcggttgcccgttct (SEQ ED NO: 7)

MOOSE00721 ctgl6228 1520153..1520285, 1521083..1521207, 1522029..1522164,

1522413..1522612, 1522847..1523221 RAVDAWLWLFFAALMLLGLVGNSLVTYVICRHKPMRTVTNFYIANLAATDV

TFLLCCVPFTALLYPLPGWVLGDFMCKFVNYIQQVSVQATCATLTAMSVDRWYVT WPLRALHRRTPRLALAVSLSIWVGSAAVSAPVLALHRLSPGPRAYCSEAFPSRALER AFALYNLLALYLLPLLATCACYAAMLP ILGRVARAGAVRAKVSRLVAAVVLLFAA CWGPIQLFLVLQALGPAGSWHPRSYAAYALKTWAHCMSYSNSALNPLLYAFLGSHF RQAFRRVCPCAPRRPP ^RJ^GPSDPAAPHAELLRLGSHPAPARAQK (SEQ ID NO:

10) cgggccgtggacgcctggctcgtgccgctcttcttcgcggcgctgatgctgctgggcctggtggggaactcgctggtcatc tacgtcatctgccgccacaagccgatgcggaccgtgaccaacttctacatcgccaacctggcggccacggacgtgaccttcctcctgt gctgcgtccccttcacggccctgctgtacccgctgcccggctgggtgctgggcgacttcatgtgcaagttcgtcaactacatccagcag gtctcggtgcaggccacgtgtgccactctgaccgccatgagtgtggaccgctggtacgtgacggtgttcccgttgcgcgccctgcacc gccgcacgccccgcctggcgctggctgtcagcctcagcatctgggtaggctctgcggcggtgtctgcgccggtgctcgccctgcac cgcctgtcacccgggccgcgcgcctactgcagtgaggccttccccagccgcgccctggagcgcgccttcgcactgtacaacctgct ggcgctgtacctgctgccgctgctcgccacctgcgcctgctatgcggccatgctgcgccacctgggccgggtcgcccgcgcaggcg ccgtgcgggccaaggtctcgcggctggtggcggccgtggtcctgctcttcgccgcctgctggggccccatccagctgttcctggtgct gcaggcgctgggccccgcgggctcctggcacccacgcagctacgccgcctacgcgcttaagacctgggctcactgcatgtc ctacagcaactccgcgctgaacccgctgctctacgccttcctgggctcgcacttccgacaggccttccgccgcgtctgcccctgcgcg ccgcgccgcccccgccgcccccgccggcccggaccctcggaccccgcagccccacacgcggagctgctccgcctggggtccca cccggcccccgccagggcgcagaag (SEQ ED NO: 9)

MOOSE00741 ctg877 402877..402934, 519710..520656

AALEKYYLSIFYGffiFVVGVLGNTIVVYGYIFSLKNWNSSNrYLFNLSVSDLAF LCTLPMLIRSYANGNWIYGDVLCISNRYVLHANLYTSILFLTFISIDRYLIIKYPFREHL LQKKEFAILISLAIWVXVTLELLPILPLINPVITDNGTTCNDFASSGDPNYNLIYSMCLT LLGFLIPLF VMCFFYYXIALFLKQRNRQVATALPLEKPLNL VTMAWIFS VLFTPYHV MRNVTUASRLGSWKQYQCTQVVINSFYIVTRPLAFLNSVINPWW^ NQLRHNFKSLTSFSRWAHELLLSFRENDSPSSPSSNGAITEVSHRT (SEQ ID NO: 12) gctgccctggaaaag actacctttccattttttatgggattgagttcgttgtgggag^ccttggaaataccattgttgtttacggc tacatcttctototgaagaactggaacagcagtaatatttatctcttlaacctctctgtctctgacttagcttttctgtgcaccctccccatgctg ataaggagttatgccaatggaaactggatatatggagacgtgctctgcataagcaaccgatatgtgcttcatgccaacctctataccagc attctctttctcacttttatcagcatagatcgatacttgataattaagtatcctttccgagaacaccttctgcaaaagaaagagtttgctatttt^ atctccttggccatttgggttttagtaaccttagagttactacccatacttccccttataaatcctgttataactgacaatggcaccacctgtaa tgattttgcaagttctggagaccccaactacaacctcatttacagcatgtgtctaacactgttggggttccttattcctcltlttg^gatgtgttt cttttattacaagattgctctcttcctaaagcagaggaataggcaggttgctactgctctgccccttgaaaagcctctcaacttggtcatcat ggcagtggtaatcttctctgtgctttttacaccctatcacgtcatgcggaatgtgaggatcgcttcacgcctggggagttggaagcagtat cagtgcactcaggtcg catcaactccttttacattgtgacacggcctttggcctttctgaacagtgtcateaaccctg cttctattttcttttg ggagatcacttcagggacatgctgatgaatcaactgagacacaacttcaaatcccttacatcctttagcagatgggctcatgaactccta ctttcattcagagaaaatgattctccttcctcaccctcctcaaatggtgcgattacagaagtgagccaccgcacg (SEQ ED NO:

11)

MOOSE00766 ctgl5378 1264081..1264142, 1351477..1351796, 1351914..1352461

NLPLQITLSAIMmLFVSFLGNL CLMWQKAAMRSAINILLASLAFADMLL AVLNMPFALVΗLTTRWIFGKFFCRVSAMFFWLFVIEGVAILLπSroRFLπVQRQDKL NPYRAKVLL -VSWATSFCVAFPLAVGNPDLQIPSPvAPQC GYTTNPGYQAYVILISL ISFFIPFLVILPFQMSroMGFKTRAFTTILILFA IVCWAPFTTYSLVATFSKHFYYQHN FFEISTWLLWLCYLKSALNPLIYYWWKKFHDACLDMMPKSFKFLPQLPGHTKRRIHF THTHTHTHTH1ΗTHTQKHM (SEQ ED NO: 14) aacttgcctcttcagatcaccctttctgctataatgatattcattctgtttgtgtcttttcttgggaacttgg tgtttgcctcatggttt accaaaaagctgccatgaggtctgcaattaacatcctccttgccagcctagcttttgcagacatgttgcttgcagtgctgaacatgcccttt gccctggtaactattcttøclacccgatggatttttgggaaattcttctgtaggg atctgctatgtttttctgg tatttgtgatagaaggagta gccatcctgctcatcattagcatagataggttccttattatagtccagaggcaggataagctaaacccatatagagctaaggttctgattgc agtttcttgggcaacttccttttgtgtagcttttcctttagccgtaggaaaccccgacctgcagataccttcccgagctccccagtgtgtgttt gggtacacaaccaatccaggctaccaggcttatgtgattttgatttctctcatttctttcttcatacccttcctggtaatactacctttccagatg agcattgacatgggctttaaaacacgtgccttcaccactattttgattctctttgctgtcttcattgtctgctgggccccattcaccacttacag ccttgtggcaacattcagtaagcacttttactatcagcacaacttttttgagattagcacctggctactgtggctctgctacctcaagtctgc attgaatccgctgatctactactggaggattaagaaattccatgatgcttgcctggacatgatgcctaagtccttcaagtttttgccgcagct ccctggtcacacaaagcgacggatacattttacacacacacacacacacacacacacacacacacacacgcacacacagaaacaca tg (SEQ ED NO: 13) MOOSE00772 ctg78 1060263..1060374, 1105772..1106011, 1106507..1107105

SLAHGi STVLVIFLAASFVGNIVLALVLQPvKPQLLQVTNP_?IFNLLVTDLLQI SLVAPWVVATS LFWPLNSHFCTALVSLTHLFAFASVNΗVVVSVDRYLSIIHPLSY PSKMTQRRGYLLLYGTWIVAILQSTPPLYGWGQAAFDERNALCSMrWGASPSYTILS VVSFIVIPLIVMIACYSVVTCAAPJlQHALLCYQCKAAKVTFIIIFSYVLSLGPYCFLAv^ AVWVDVΕTQVPQWVITIJIWLFFLQCC PYVΥGYMHKπKKEIQDMLKKQLNQQSS SDSGSSSSSEDERPMRSHVKNGEVGRRRRH (SEQ ED NO: 16) agcctggcccacggcatcatccgctcaaccgtgctggttatcttcctcgccgcctctttcgtcggcaacatagtgctggcgct agtgttgcagcgcaagccgcagctgctgcaggtgaccaaccgttttatctttaacctcctcgtcaccgacctgctgcagatttc gctcgtggccccctgggtggtggccacctctgtgcctctcttctggcccctcaacagccacttctgcacggccctggttagcctcaccc acctgttcgccttcgccagcgtcaacaccattgtcgtggtgtcagtggatcgctacttgtccatcatccaccctctctcctacccgtccaa gatgacccagcgccgcggttacctgctcctctatggcacctggattgtggccatcctgcagagcactcctccactctacggctggggc caggctgcctttgatgagcgcaatgctctctgctccatgatctggggggccagccccagctacactattctcagcgtggtgtccttcatc gtcattccactgattgtcatgattgcctgctactccgtggtgttctgtgcagcccggaggcagcatgctctgctgtgctaccagtgcaaag ctgctaaagtgatcttcatcatcattttctcc gtgctatccctggggccctactgctttttagcagtcctggccgtgtgggtggatgtcga aacccaggtaccccagtgggtgatcaccataatcatctggcttttcttcctgcagtgctgcatccacccctatgtctatggctacatgcac aagaccattaagaaggaaatccaggacatgctgaagaaacaactaaaccagcagtcttcatctgactctggctcctcctcctcttcaga agatgaacgacccatgagatcccatgtgaagaatggtgaggttggcaggcggcggagacat (SEQ ED NO: 15) MOOSE00775 ctgl5540 14488015..14488239, 14488744..14488768,

14488985..14489082, 14491445..14491621, 14495389..14495435, 14499020..14499178, 14539721..14539940

PGRAKMALVLTGVLIFALALFGNALVFYVv^'TRSKTMRTVTNIFICSLALSDML ITLFCIPVTMLQMSDNWLGGAFICKMVPFVQSTAVVTEILTMTCIAVERHQGLVHPF KMKWQYTNRTiAFTMLGVNWLVAVIVGSPMWHIKYDFLYEKEHICCLEEWTSPVHQ KIYTTFIivTLFLLPLMVMLILYSKIGYELWIKKRKKRAVTMMVTVVALFAVCWAPFH VVHMMffiYSEFLSHVTPSMIFAINQIIGFSNSICNP ^AFMNENFKKNVLSAVCYCIV NKTFSPAQRHGNSGITMMRKKAKFSLRENPVEET (SEQ ED NO: 18) ccgggacgcgccaagatggccctcgtgctcaccggcgtgctcatattcgccctggcgctctttggcaatgctctggtgtttta cgtggtgacccgcagcaagaccatgcgcaccgtcaccaacatctttatatgctccctggcgctcagtgacatgctcatcaccctcttttg cattcccg^caccatgctccagaacatttccgacaactggctggggggtgctttcatttgcaagatggtgccatttgtccagtctaccgct gttgtgacagaaatcctcactatgacctgcattgctgtggaaaggcaccagggacttgtgcatccttttaaaatgaagtggcaatacacc aaccgaagggctttcacaatgctaggtgtggtctggctggtggcagtcatcgtaggatcacccatgtggcacatcaaatatgacttccta tatgaaaaggaacacatctgctgcttagaagagtggaccagccctgtgcaccagaagatctacaccaccttcatccttgtcatcctcttc ctcctgcctcttatggtgatgcttattctgtacagtaaaattggttatgaactttggataaagaaaagaaagaaacgagctgtcattatgatg gtgacagtggtggctctctttgctgtgtgctgggcaccattccatgttgtccatatgatgattgaatacagtgaattcctctcccatgttactc cttctatgatttttgctatcgtgcaaattattggattttccaactccatctgtaatcccattgtctatgcatttatgaatgaaaacttcaaaaaaaa tgttttgtctgcagtttgttattgcatagtaaataaaaccttctctccagcacaaaggcatggaaattcaggaattacaatgatgcggaaga aagcaaagttttccctcagagagaatccagtggaggaaacc (SEQ ED NO: 17)

MOOSE00779 ctgl6537 5292218..5292340, 5438500..5438551, 5450751..5450956,

5450981..5451284, 5462460..5462773

FFAAKIVJGIALAGIMLVCGIGNF IAALTRYKKLRNLTNLLIANLAISDFLVA ΠCCPFEMDYYVVRQLSWEHGHVXCASVNYLRTVSLYVSTNALLAIAIDRYLAIVHPL KPRMNYQTASFLIALVWMVSILIAIPSAYFATETVLFIVKSQEKIFCGQIWPVDQQLY YKSYFLFIFGVEFVGPVVTMTLCYAMSRELWFKALRKRLRCRRKTVLVLMCILTAYV LCWAPFYGFΗVPJ⁾FFPTWVKEKHYLTAFY\^VECLAMSNSMINTVPFLYFWLRWGF TIFLRLTLSQKKKKRRRRKRKRTESRDSNRYLYTHVPSSNHSTPAW (SEQ ED NO: 20) ttcttcgcagccaagatcgtcattggcattgcactggcaggcatcatgctggtctgcggcatcggtaactttgtctttatcgctg ccctcacccgctataagaagttgcgcaacctcaccaatctgctcattgccaacctggccatctccgacttcctggtggccatcatctgct gccccttcgagatggactactacgtggtacggcagctctcctgggagcatggccacgtgctctgtgcctccgtcaactacctgcgcac cgtctccctctacgtctccaccaatgccttgctggccattgccattgacagatatctcgccatcgttcaccccttgaaaccacggatgaatt atcaaacggcctccttcctgatcgccttggtctggatggtgtccattctcattgccatcccatcggcttactttgcaacagaaacggtcctc tttattgtcaagagccaggagaagatcttctgtggccagatctggcctgtggatcagcagctctactacaagtcctacttcctcttcatcttt ggtgtcgagttcgtgggccctgtggtcaccatgaccctgtgctatgccaggatctcccgggagctctggttcaaggcaattcgcaagc ggctgcgctgccgcaggaagacggtcctggtgctcatgtgcattctcacggcctatgtgctgtgctgggcacccttctacggtttcacc atcgttcgtgacttcttccccactgtgttcgtgaaggaaaagcactacctcactgccttctacgtggtcgagtgcatcgccatgagcaaca gaagaagaagaaaaagaaaaagaactgaaagcagagactcaaacagatatttgtacactcatgttcctagcagcattattcacagcact ccagc ctgg (SEQ ED NO: 19)

MOOSE00804 ctg30162 16180..16299, 37790..38066, 71494..71629, 82561..82975

SWELQMFFFM SLLYVATMVGNSLIVITVΓVDPHLHSPMYFLLTNLSΠDMSL ASFATPKMTDYLTGHKTISFDGCLTQIFFLHLFTGTEΠLLMAMSFDRYIAICKPLHYA SVISPQVCVALWASWIMGVMHSMTPFIEYLLIRVSFLHCLSLLTLLKNSWLQVRGF GFCILFYCLSFTFSLLVSSYΠILVTVWLKSSAAMAKAFSTLASHIAVVTLFFGPCIFI

YV VPFΗSPLDK_^?LAIFYT TPVXNPIIYTLRlNuΗ)MKAAVRXm KNSAKKTTNTKRTLDHVDLPDSHRT (SEQ ED NO: 22) tcctgggaactacagatgtttttctttatgg g tttcattgctttatgtggcaacaatggtgggtaacagcctcatagtcatcaca gttatagtggaccctcacctacactctcctatgtatttcctgcttaccaatctttcaatcattgatatgtctcttgcttctttcgccaccccaaag atgattacagattacctaacaggtcacaaaaccatctcttttgatggctgccttacccagatattctttctccaccttttcactggaactgaga tcatcttactcatggccatgtcctttgataggtatattgcaatatgcaagcccctgcactatgcttctgtcattagtccccaggtgtgtgttgct ctcgtgg ggcttcctggattatgggagttatgcattcaatgacaccatttattgaatatttattgattagagtgtcctttctccattgcttgtctt tgftgactttactaaagaacagctggttacaggtacgtggctttgtttttggg tcagctcctacatcattattcttgttacagtttggctcaagtcttcagctgcaatggcaaaggcattttctacgctggcttcccatattgcagt agtaatattattctttggaccttgcatcttcatctatgtgjggccctttaccatctrt^ ccccgtcctaaaccccattatttatacactaaggaatagggatatgaaggctgccgtaaggaaaattaggacttcaatatctcactttcgg caaaggacagatcatctagacaaaaattcagccaaaaaaacaacaaatacaaagcgcactctagatcatgtggacctaccagatagtc acagaaca (SEQ ED NO: 21)

MOOSE00814 ctgl5378 3973801..3973891, 3975545.-3975625, 4023376..4023510,

4032595..4032652, 4060801..4060926, 4186201..4186358, 4227631..4227840, 4235835..4235929 SVVTJTVILPSMGΠCSTGLVGMLIWTIIRSR__ :TWDIYICNLAVADLVHIVG

MPFLFFLQWARGGEWWGGPLCTΠTSLDTCNQFACSA TVMSVDRYFALVQPFRLT RWRTRYKTIRJJSΓLGLWAASFILALPVWVYSKVIKFKDGVESCLSNLLIFSKKYFISLIF CΠFFISISFISALMTFLLLFLEKQQQQKKKHTSQVMHLLLLSSTFLVSWIPRYFILFEAI VNGSSLMIWTSVCLLLVYK CDFCTIJRYFDVKCWRRNYR TESLSSTPTQHRKVWFGLARPLWAT (SEQ ED NO: 24) agtgtggtagatacagtcatcctcccttccatgattgggattatctgttcaacagggctggttggcaacatcctcattgtattcac tataataagatccaggaaaaaaacagtccctgacatctatatctgcaacctggctgtggctgatttggtccacatagttggaatgccttttc ttattcaccaatgggcccgagggggagagtgggtgtttggggggcctctctgcaccatcatcacatccctggatacttgtaaccaatttg cctgtagtgccatcatgactgtaatgagtgtggacaggtactttgccctcgtccaaccatttcgactgacacgttggagaacaaggtaca agaccatccggatcaatttgggcctttgggcagcttcctttatcctggcattgcctgtctggg^ctactcgaaggtcatcaaatttaaagac ggtgttgagagttgtttgtcaaatttgcttatcttttcaaagaaatattttatttctttgatottttgtattat^ ctgatctttattacatttcttctactattcttagaaaaacaacaacaacaaaaaaaaaaacacacgtcacaggtcatgcatctcctactcttgt cttccacattccttgtaagttggattcctaggtattttattctctttgaagcaattgtgaatgggagttcactcatgatttggctctctgt^ gttattggtgtataagaatgcttgtgatttttgcaccattcattattttgatgttaaatgtg^ctttaggagaaattacagacaattgtcaaagaa aaaaagccatggaagctttgtgcccacaacccaaacagaaagtttatoctccaccccaacccaacacaggaaagtggttgtttttggttt ggccagaccactatgggctact (SEQ ED NO: 23)

MOOSE00818 ctgl5907 32727057..32727450, 32800163..32800610, 32874979..32875093

WPHLEWLFWILIFYLITLIGNLFIΠLSYLDSHLHTPMYFFLSNLSFLDLCYTT

SSIPQLLVNLWGPEKTISYAGCTVQLYFVLALGTAECVLLWMSYDRYAAVCRPLH

YTVLMHPRFCRLLAAASWVTLHSSFTFWWLCGHRLVTJHFFCEWALLRLSCVDTH

A>_ΪLTLMVMSSIFVLIPLILILTAYGAIARAVLSMQSTTGLQKVFRTCGAHLMVVSLF FIPVMCMYLQPPSENSPDQGKFIALFYT\^TPSLNPLΓY LR>___VKGAAKRLLGACR

PMQSQPRTVRSSFFDRKATGCEPGSCVRTPAAQTC (SEQ ED NO: 26) tggcctcatctggaagtagttctctttgtggttatcttgatcttctacttgataacactgataggaaacctgttcatcatcatcctgt catacctggactcccatctccacactcccatgtacttcttcctttcaaatctctcatttctggatctctgctacaccaccagctctatccctca gttgctggtgaatctctggggcccggaaaagaccatctcttatgctggttgtacagttca actttactttgttctcgcactgggaaccgcagagtgtgtcctactggtggtgatgtcctatgatcgttatgcagctgtgtgtagacctttgca ttacactgtcctcatgcaccctcgtttctgccgcttgttggctgcggcttcttgggtaacac^^ tggacatcgcctagtggatcacttcttctgtgaagttccagcacttctgcgtttatcatgtgttgacacccatgcaaatgagctgaccctcat ggtcatgagctccatttttgttctcatacctctcattctgattctcactgcctatggtgccattgcccgggctgtactgagcatgcaatcaacc actgggcttcagaaagtgtttaggacatgtggagcccatcttatggttgtatctctctttttcattccagtcatgtgcatgtatctccagccac catcagaaaattctcctgatcagggcaagttcattgccctcttttatactgttgtcacaccgagtcttaatcctctaatctacactctcagaaa caagcatgtaaaaggggcagcgaagagactattgggtgcctgccgccccatgcagagccagcccagaactgtgaggtcaagtttctt tgatagaaaggcaacaggttgtgagcctggctcctgtgtgaggactccggcagcacagacttgt (SEQ ED NO: 25)

MOOSE00822 ctgl4294 995965..996055, 1019834..1020641, 1219431..1219494 VRVVPRTWFLFFFLFILVLVSMLTVΠLSQLVARRQKSSYNYLLALAAADILV

LFFIVF\T)FLLEDFILNMQMPQ DKIIEVLEFSSFFLTSIWITVPLTRORYIAVCHPLKYH TVSYPARTRKVIVSVYITCFLTSIPYYWWPNIWTEDYISTSVHHVLIWFFLCFTVYLVPC SIFFILNSIIVYKLRRKSNFRLRGYSTGKTTAILFTITSIJATLWAPPJIMILYHLYGAPIQ NRWLVHMSDIANMLALLNTAINFFLYCFISKRFRTMAAATLKAFFKCQKQPVQIQPF KQISLSPTHTHTHTHTHTHTHTSKDT (SEQ ED NO: 28) gttcgtgtcgtgccccgcaccgtgtULltLtgtttlttlttLlgttLattttggttcttgtttcaaatatcttgacagtgatcatcctctcc cagctggtggcaagaagacagaagtcctcctacaactatctcttggcactcgctgctgccgacatcttggtcctctttttcatagtgtttgt ggacttcctgttggaagatttcatcttgaacatgcagatgcctcaggtccccgacaagatcatagaagtgctggaattctcatccatccac acctccatatggattactgtaccgttaaccattgacaggtatatcgctgtctgccacccgctcaagtaccacacggtctcatacccagccc gcacccggaaagtcattgtaagtgtttacatcacctgcttcctgaccagcatcccctattactggtggcccaacatctggactgaagacta catcagcacctctgtgcatcacgtcctcatctggatccactgcttcaccgtctacctggtgccctgctccatcttcttcatcttgaactcaat cattgtgtacaagctcaggaggaagagcaattttcgtctccgtggctactccacggggaagaccaccgccatcttgttcaccattacctc catctttgccacactttgggccccccgcatcatcatgattctttaccacctctatggggcgcccatccagaaccgctggctggtacacatc atgtccgacattgccaacatgctagcccttctgaacacagccatcaacttcttcctctactgcttcatcagcaagcggttccgcaccatgg cagccgccacgctcaaggctttcttcaagtgccagaagcaacctgtacaaattcaacctttcaagcagatatctttaagtccaacacaca cacacacacacacacacacacacacacacacacacacttctaaagacact (SEQ ED NO: 27)

MOOSE00826 ctgl5968 677807..678430, 679388..679470, 683881..684142

MGPGEALLAGLLVMVLAVALLSNALVLLCCAYSAELRTRASGVLLVNLSLG HLLLAALDMPFTLLGVMRGRTPSAPGACQVIGFLDTFLASNAALSVAALSADQWLA VGFPLRYAGRLRPRYAGLLLGCAWGQSLAFSGAALGCSWLGYSSAFASCSLRLPPEP ERPP^AAFTATLHAVGFVLPLAVLCLTSLQVHRVARRHCQRMDTRRRHRATRKIGIA IATFLICFAPYVMTRLAJiL FVTVNAQWGILSKCLTYSKAVADPFTYSLLRRPFRQV LAGMVFTRLLKRTPPJ'ASTHDSSLDVAGMVHQLLKRTPRPASTHN (SEQ ED NO: 30) atgggccccggcgaggcgctgctggcgggtctcctggtgatggtactggccgtggcgctgctatccaacgcactggtgct gctttgttgcgcctacagcgctgagctccgcactcgagcctcaggcgtcctcctggtgaatctgtctctgggccacctgctgctggcgg cgctggacatgcccttcacgctgctcggtgtgatgcgcgggcggacaccgtcggcgcccggcgcatgccaagtcattggcttcctgg acaccttcctggcgtccaacgcggcgctgagcgtggcggcgctgagcgcagaccagtggctggcagtgggcttcccactgcgctac gccggacgcctgcgaccgcgctatgccggcctgctgctgggctgtgcctggggacagtcgctggccttctcaggcgctgcacttggc tgctcgtggcttggctacagcagcgccttcgcgtcctgttcgctgcgcctgccgcccgagcctgagcgtccgcgcttcgcagccttca ccgccacgctccatgccgtgggcttcgtgctgccgctggcggtgctctgcctcacctcgctccaggtgcaccgggtggcacgcagac actgccagcgcatggacacccggcgccgccaccgcgccaccaggaagattggcattgctattgcgaccttcctcatctgctttgcccc gtatgtcatgaccaggctggcggagctcgtgcccttcgtcaccgtgaacgcccagtggggcatcctcagcaagtgcctgacctacag caaggcggtggccgacccgttcacgtactctctgctccgccggccgttccgccaagtcctggccggcatggtgcaccggctgctgaa gagaaccccgcgcccagcatccacccatgacagctctctggatgtggccggcatggtgcaccagctgctgaagagaaccccgcgc ccagcgtccacccacaac (SEQ ED NO: 28)

MOOSE00829 ctgl4145 39703..40567, 63946..64061 LEGIKHWIFiPFFFMYMVAISGNCFILIIIKTNPRLHTPMYYLLSLLALTDLGLC

VSTLPTTMGIFWFNSQSIYFGACQIQMFCIHSFSFMESSVLLMMSFDRFVAICHPLRYS VIITGQQVΥRAGLIVTFRGPVAΗPIVLLLKAFPYCGSVVLSHSFCLHQEVIQLACTDTT Iτ πSu-YGLMVV TVMLDLVL_ LSYGLιLHTVAGLASQEEQP_lAFQTCTAHLCAVL FWMMGLSLVΗRFGKHAPPAfflLLMANVYLF PMLNPIIYSIKTKEIHRAIIKLLE CRSLRSQCNQLEERVSVMEDEMNEMKQEEKFREKRIKR (SEQ ED NO: 32) ttggaaggcatcaaacactggattttcatcccctttttctttatg^catggttgccatctcaggcaattgtttcattctgatcattatt aagaccaaccctcgtctgcacacacccatgtactatctactatccttgctggccctcactgacctggggctgtgtgtgtccacgttgccca ccactatggggatcttctggtttaactcccagagtatctactttggagcgtgtcaaatccagatgttctgcatccactctttttccttcatgga gtcctcagtgctcctcatgatgtcctttgaccgctttgtggccatctgccaccctctgaggtattcggtcattatcactggccagcaagtgg tcagagcaggcctaattgtcatcttccggggacctgtggccactatccctattgtcctcctcctgaaggcttttccctactgtggatctgtg gtcctctcccactcattttgcctgcaccaggaagtgatacagctggcctgcacagataccaccttcaataatctgtatggactgatggtgg tagttttcactgtgatgctggacctggtgctcatcgcactgtcctatggactcatcctgcacacagtagcaggcctggcctcccaagagg agcagcgccgtgcctttcagacatgcaccgctcatctctgtgctgtgctagtattctttgtgcccatgatggggctgtccctggtgcaccg ttttgggaagcatgccccacctgctattcatcttcttatggccaatgtctacctttttgtgcctcccatgcttaacccaatcatatacagcatta agaccaaggagatccaccgtgccattatcaaactcctagaatgcagaagtctcaggagccaatgcaatcaactggaagaaagggtat cagtgatggaagatgaaatgaatgaaatgaagcaagaagagaagtttagagaaaaaagaataaaaaga (SEQ ED NO: 31)

MOOSE00838 ctgl4667 1435690..1435800, 1536739..1536780, 1544674..1544727,

1594182..1594286, 1597404..1597553, 1597578..1598087 HSTADLVLFSWMAWWALCGNVLLIFLIYMDPHLHTPMYFFLSQLSLMDL

MLVCTNWKMAANFLSGRKSISFVGCGIQIGLFVCLVGSEGLLLGLMAYDRYVAISH PLHYPILMNQRVCLQITGSSWAFGimGLIQMVVVMNFPYCGLRKVNHFFCEMLSLL KLACWACC MLLFPFSIWASYAFflLGTVLQMHSAQAWKKALATCSSHLTAFFFC QLPYQILSHQVVCSFYWvTLGHRCDKSSLQSSTVSLSFAVLNPILYGSVARSFRRRAG A LLVCRKKPQNSSENFTFTPTSFLLSPYITHTHTHTHAHTHV (SEQ ED NO: 34) cacagtactgctgaccttgtcctcttctccgtggttatggcggtcttcacagtggccctctgtgggaatgtcctcctcatcttcct catctacatggaccctcaccttcacacccccatgtacttcttcctcagccagctctccctcatggacctcatgttggtctgtaccaatgtgc caaagatggcagccaacttcctgtctggcaggaagtccatctcctttgtgggctgtggcatacaaattggcctctttgtctgtcttgtggga tctgaggggctcttgctgggactcatggcttatgaccgctatgtggccattagccacccacttcactatcccatcctcatgaatcagagg gtctgtctccagattactgggagctcctgggcctttgggataatcgatggcttgatccagatggtggtagtaatgaatttcccctactgtgg cttgaggaaggtgaaccatttcttctgtgagatgctatccttgttgaagctggcctgtgtgatatttgcttgctgtgtcttcatgcttctcttccc attctccatcatcgtggcctcctatgctcacattctagggactgtgctgcaaatgcactctgctcaggcctggaaaaaggccctggccac ctgctcctcccacctgacagctttttLLtlltgccaactaccttøccaaatcctttoccatoaagttgtctgttcattttattggg gattttgggg catagatgtgacaaatettcattgcaatcaag^cagtgtcactttcatttgcggttcttaatccaattctatatggcagcgtcgcccgctcct ttcggcgcagggcgggagcccttcttgtgtgcagaaaaaaaccacagaatagctcagaaaatttcacttttacccccacttctttcctcct ttctccttacatcacgcacacacacacacacacacatgcacacacacacgtc (SEQ ED NO: 33)

MOOSE00843 ctgl5064 31780507..31780654, 31841187..31841254, 31873514..31873564, 31907740..31907833, 31923371..31923572, 31930354..31930411, 31940910..31940952, 31944453..31944599, 32115413..32115576

LCFLQTEQLITLWVLFWTΓV^GNSVVXFSTWRRKKKSRMTFFVTQLAITVGFL LDFLILFMPLHFSLVΥLFHYESSPDFWCLQSYFFCV T,LYASTYV VSLSRORYHAΙV YPMKFLQGEKQARVLIVIAWSLSFLFSIPTLIIFGKRTLSNGEVQCWALWPDDSYWTP YMTIVAFLVYFIPLTπSIMYGIVIRTIWrc^

DILDNFNLLPDTQEPJ^YASVΠQNLPALNSAINPLIYΎSVSQKNKRKRKRRSRRKKKE KERRRRKKRRRRKKKEGRRGGEGRGKKEEERRKE (SEQ ED NO: 36) ctctgtttcttgcagactgagcaattgataactctgtggg cctctttg trttaccattg tggaaactccgttgtgcttttttccaca tggaggagaaagaagaagtcaagaatgaccttctttgtgactcagctggccatcacagtgggatttttgttggactttctcatcctattcatt atgcctcttcacttctctttggtctatctctttcactatgagagctcacrt^ tgctgctctacgcctctacctacgtcctggtgtccctcagcatagacagataccatgccatcgtctaccccatgaagttccttcaaggaga aaagcaagccagggtcctcattgtgatcgcctggagcctgtcttttctgttctccattcccaccctgatcatatttgggaagaggacactgt ccaacggtgaagtgcagtgctgggccctgtggcctgacgactcctactggaccccatacatgaccatcgtggccttcctggtgtacttc atccctctgacaatcatcagcatcatgtatggcattgtgatccgaactatttggattaaaagcaaaacctacgaaacagcaaaaatcaag gctatcaagtatagcatcatcatcattcttgccttcatctgctgttggagtccatacttcctgtttgacattttggacaatttcaacctccttcca gacacccaggagcgtttctatgcctctgtgatcattcagaacctgccagcattgaatagtgccatcaaccccctcatctactactctgtttc tcaaaaaaacaaaagaaagagaaaaagaagaagtaggaggaagaagaaggaaaaagaaagaagaagaaggaagaagagaagg agaaggaagaagaaagaaggaagaagaggaggagaaggaagaggaaagaaggaagaagagaggaggaaggaa (SEQ ED NO: 35)

MOOSE00846 ctgl5944 960216..961037, 988991..989137

NLELWKIFSAWLVMYVATVLENLLΓVΎTΠTSQSLRSPMYFFLTFLSLLDVMF SSVVAPKVIVTJTLSKSTTISLKGCLTQLFVEHFFGGVGIILLTVMAYDRYVAICKPLHY TIIMSPRVCCLMVGGAWVGGFMHAMQLLFMYQIPFCGPNIΓDHFICDLFQLLTLACT DTFFLLGLLVTLNSGMMCVAIFLILIASYTVTLCSLKSYSSKGRHKALSTCSSHLTVVVL FFVPCIFLYMRPVVTHPROKAMAVSDSΠTPMLNPLIYTLRNAEDIREILRLMFRAPFCP LFYFIFPGWSQNPELRSPSSASQSSGVTGVSHC (SEQ ED NO: 38) aacctggagctgtggaaaatattttctgctgtgtttcttgtcatgtatgtagccacagtgctggaaaatctacttattgtggtaact attatcacaagtcagagtctgaggtcacctatgtatttttttcttaccttottgtcccttttggatgtcatgttctcatctgtcgttgcccccaagg tgattgtagacaccctctccaagagcactaccatctctctcaaaggctgcctcacccagctgtttgtggagcatttctttggtggtgtggg gatcatcctcctcactgtgatggcctatgaccgctacgtggccatctgtaagcccctgcactacacgatcatcatgagtccacgggtgtg ctgcctaatggtaggaggggcttgggtggggggatttatgcacgcaatgatacaacttctcttcatgtatcaaatacccttctgtggtccta atatcatagatcactttatatgtgatttgtttcagttgttgacacttgcctgcacggacacccacatcctgggcctcttagttaccctcaacag tgggatgatgtgtgtggccatctttcttatcttaattgcgtcctacacggtcatcctatgctccctgaagtcttacagctctaaagggcggca caaagccctctotacctgcagctcccacctcacggtggttgtattgttctttgtcccctgtattttcttgtacatgaggcctgtggtcactcac cccatagacaaggcaatggctgtgtcagactcaatcatcacacccatgttaaatcccttgatctatacactgaggaatgcagaggacata agagagatactgaggctgatgtttcgtgcccccttttgtcctttattttattttatttttccaggctggtctcaaaatcctgagctcaggtcgcc ctcctcggcctcccaaagttctggagttacaggtgtgagccactgc (SEQ ID NO: 37) MOOSE00855 ctgl4333 2003002..2003122, 2024409..2025211

VFfTAYLVLSSLAMFTCLCGMAGNSMVTWLLGFRMHRNPFCIYILNLAAADLL FLFSMASTLSLETQPLVNTTDKVHELMKRLMYFAYTVGLSLLTAISTQRCLSVLFPIW FKCHRPRHLSAWVCGLLWTLCLLMNGLTSSFCSKFLKFNEDRCFRVDMVQAALIMG VLTPVMTLSSLTLFVWVRRSSQQWRRQPTRLFVVVLASVLVFLICSLPLSIYWFVLY WLSLPPEMQVXCFSLSRLSSSVSSSANPVIYFLVGSRRSHRLPTRQGRCLTLSTRFREN SITRTAPRGKSTPRIQSPSTRPHILQ (SEQ ED NO: 40) gtgcacacggcctacctggtgctgagctccctggccatgttcacctgcctgtgcgggatggcaggcaacagcatggtgatc tggctgctgggctttcgaatgcacaggaaccccttctgcatctatatcctcaacctggcggcagccgacctcctcttcctcttcagcatgg cttccacgctcagcctggaaacccagcccctggtcaataccactgacaaggtccacgagctgatgaagagactgatgtactttgcctac acagtgggcctgagcctgctgacggccatcagcacccagcgctgtctctctgtcctcttccctatctggttcaagtgtcaccggcccag gcacctgtcagcctgggtgtgtggcctgctgtggacactctgtctcctgatgaacgggttgacctcttccttctgcagcaagttcttgaaat tcaatgaagatcggtgcttcagggtggacatggtccaggccgccctcatcatgggggtcttaaccccagtgatgactctgtccagcctg accctctttgtctgggtgcggaggagctcccagcagtggcggcggcagcccacacggctgttcgtggtggtcctggcctctgtcctgg tgttcctcatctgttccctgcctctgagcatctactggtttgtgctctactggttgagcctgccgcccgagatgcaggtcctgtgcttcagct tgtcacgcctctcctcgtccgtaagcagcagcgccaaccccgtcatctacttcctggtgggcagccggaggagccacaggctgccca ccagacaggggcggtgcctcacactgtcaacccgattccgtgagaactctatcacgagaacagcaccacgggggaaatccaccccc aggatccaatcaccttccaccaggccacacatcctacaa (SEQ ED NO: 39) MOOSE00861 ctgl6279 54051..54182, 115058..115808, 116858..116925

PCVAGVIPVIYYSVLLGLGLPGDLLTAVALARLATRTRRPSYYYLLALTASDΠ IQVVTV^AGFLLQGAVLARQVPQAVVRTAN LEFAANHASVWIAILLTVDRYTALCH PLHHRAASSPGRTRRAIAAVLSAALLTGIPFYWWLDMWRDTDSPRTLDEVLKWAHC LTVYFIPCGVFLVTNSAIIHRLRRRGRSGLQPRVGKSTAILLGITTLFTLLWAPRVFVM LYHMYVAPVHRDWRVHLALDVAISΠVIVAMLHTAANFGLYCFVSKTFRATFCSCCPS CSAMGRSQLTATSSSRVQAVLLPQPPEELGLQARATT (SEQ ED NO: 42) ccgtgtgtggctggcgtcatccctgtcatctactacagtgtcctgctgggcttggggctgcctggtgacctcctgaccgcagt ggccctggcgcgccttgccaccaggaccaggaggccctcctactactaccttctggcgctcacagcctcggatatcatcatccaggtg gtcatcgtgttcgcgggcttcctcctgcagggagcagtgctggcccgccaggtgccccaggctgtggtgcgcacggccaacatcctg gagtttgctgccaaccacgcctcagtctggatcgccatcctgctcacggttgaccgctacactgccctgtgccaccccctgcaccatcg ggccgcctcgtccccaggccggacccgccgggccattgctgctgtcctgagtgctgccctgttgaccggcatccccttctactggtgg ctggacatgtggagagacaccgactcacccagaacactggacgaggtcctcaagtgggctcactgtctcactgtctatttcatcccttgt ggcgtgttcctggtcaccaactcggccatcatccaccggctacggaggaggggccggagtgggctgcagccccgggtgggcaaga gcacagccatcctcctgggcatcaccacactgttcaccctcctgtgggcgccccgggtcttcgtcatgctctaccacatgtacgtggcc cctgtccaccgggactggagggtccacctggccttggatgtggccaatatggtggccatgctccacacggcagccaacttcggcctct actgctttgtcagcaagactttccgggccactttttgctcttgttgcccaagctgtagtgcaatggggcgatctcagctcactgcaacctcc tcctcccgggttcaagcagttctcctgcctcagcctcccgaggagctgggactacaggcgcgtgccaccaca (SEQ ED NO: 41) MOOSE00872 ctgl8147 32565..32680, 45290..45647, 45666..46148

DPELKLIPFSLFLSMYLVTILGNLLILLAVISDSHLHTPMYFLLFNLSFTDICLTT TTVPKILVNIQAQNQSITYTGCLTQICLVLVFAGLESCFLAVMAYDRYVAICHPLRYT VLMNVHFWGLLILLSMFMSTMDALVQSLMVLQLSFCKNVEIPLFFCELACSDTLINN ILIY ASSWGAIPLSGIIFSYSQIVTSVLP_ PSARGKYKAFSTCGCHLSVFSLFYGTAF GVYISSAVAESSmTAVASVMYTVNPQMMOTFIYSLPNKEMKKALRKLIECLTLCFV LFFSLRWSLSLSPRLECNGTTSAHCNFRLRV (SEQ ED NO: 44) gacccagaactgaagttaatccctttcagcctgttcctgtccatgtacctggtcaccatcctggggaacctgctcattctcctg gctgtcatctctgactcccacctccacacccccatgtacttccttctctttaatctctcctttactgacatctgtttaaccacaaccacagtccc aaagatcctagtgaacatccaagctcagaatcagagtatcacttacacaggctgcctcacccagatctgtcttgtcttggtttttgctggct tggaaagttgctttcttgcagtcatggcctacgaccgctatgtggccatttgccacccactgaggtacacagtcctcatgaatgtccatttc tggggcttgctgattcttctctccatgttcatgagcactatggatgccctggttcagagtctgatggtattgcagctgtccttctgcaaaaac gttgaaatccctttgttcttctg gaactcgcctgttctgacaccctcatcaacaacatcctcatatattttgcaagtagtgtatttggtgcaatt cctctctotggaataattttctcttattctcaaatagtcacctctgttctgagaatgccatcagcaagaggaaagtataaagcgttttccacct gtggctgtcacctctctgttttttccttgttctatgggacagcttttggggtg acattagttctgctgttgctgagtcttcccgaattactgctgt ggcttcagtgatgtacactgtggtccctcaaatgatgaaccccttcatctacagcctgagaaataaggagatgaagaaagctttgagga aacttattgaatgcctgactctttgttttgltttgtlltltLctctgagatggagtctttctctgtctcccaggctggagtgcaatggcacgacctc ggctcactgcaacttccgcctccgggtt (SEQ ED NO: 43) MOOSE00880 ctgl5944 3962338.3962443, 4014447..4014532, 4029024..4029117,

4033619..4034316

ATEFQVLLFLLFLLLYLMILCGNTAITWVVCTHSTLRTPMYFFLSNLSFLELCY TTVVΥPLMLSMLGAQ ISLAGCGAQMFFFVTLGSTDCFLLAIMAYDRYVAICHPL HYTLMTRELCTQMLGGALGLALFPSLQLTALIFTLPFCGFfflQEINHFLCDVPPVLRL ACADIJΛVHQAVLYVNSILVLTIPFLLICVSYVFITCAILS SAEGP lRAFSTCSFHLTV VLLHFPPLQVHHFFKNSAPFKTLLIPFIQPFGYVNVIPMLNPLIYSLRNKEVKEALRKIL NRAKTQ VTQTHRETGSHIHRQTDGCRPRQTGRYLHRQTE (SEQ ED NO: 46) gccactgaattccaggttcttctcttccttctcttcctcctcctctacttgatgatcctctgtggcaacacagccatcatctgggtg gtgtgcacacacagcaccctccgcaccccgatgtatttcttcctgtccaacctgtctttcctggaactctgctacaccaccgtggtagtac ccttgatgctttccaacattttgggggcccagaagcccatttcgttggctggatgtggggcccaaatgttcttctttgtcaccctcggcagc acggactgtttcctcttggcgatcatggcctatgaccgctatgtggctatctgccacccgctgcactacaccctcatcatgacccgcgag ctgtgcacgcagatgctgggtggggccctgggcctggccctcttcccctccctgcagctcaccgccttaatcttcaccctgcccttttgc ggccaccaccaggaaatcaaccacttcctctgcgatgtgcctcccgtcctgcgcctggcctgcgctgacatccgcgtgcaccaggct gtcctctatgtcgtgagcatcctcgtgctgaccatccccttcctgctcatctgcgtctcctacgtgttcatcacctgtgccatcctgagcatc cgttctgccgagggccgccgccgggccttctccacctgctccttccacctcaccgtggtcctgctgcactttc ctcccctgcagg tcaccacttctttaaaaattctgctcctttcaaaaccctgctcattcccttcatccagccatttgggtatgtgaatgtcatc cccatgttgaatcccctcatctacagcctgaggaacaaggaagtgaaggaggccctgagaaaaattctcaatagagccaagacacag gtgacacagacacatagagagactggtagtcacatacacaggcagacagatgggtgcaggcctaggcagacaggcagatatctgc atagacagacagaa (SEQ ED NO: 45)

MOOSE00882 ctgl5944 1984917..1985024, 2124131..2124428, 2127756..2127837,

2273894..2274344

DPQMEIIFFVVFLIVYLVNWGNIGMIILITTDTQLHTPMYFFLCNLSFVDLGYS SAIAPRMLADFLTNHKVISFSSCATQFAFFVGFVDAECYVLAAMAYGRFVAICRPLH YSTFMSKQVCLALMLGSYLAGLVSLVAHTTLTFSLSYCGCCIYLKERAGHRKLNYSI FFILFSLFFSLIi iSYIFILIAILRMRSAESRRKAFSTCGSHLVAVTVFYGTLFCMYVT^ PTDRSVEQSK\TA YTFVSPMLNPIIΥSLP_^KDVKQAFWKLIRRNKSMACGRVGKT KCSERPEKDPSICSDSEΓVAAWKE (SEQ ID NO: 48) gaccctcagatggagatcatcttcttcgtggtcttcctcatagtttacctggttaatgtagtggggaatattggtatgattatcctg attacaacagacactcagcttcacacacccatgtattttttcctctgcaacctctcctttgttgacctgggctactcctcagccattgccccc aggatgctggctgacttcctaacaaatcacaaagttatctccttctccagctg^gccacccagtttgctttttttgtaggttttgtggatgctg agtgctatgtcctggcagccatggcctatggtcgttttgtggccatttgtcgacccctccactatagcaccttcatgtccaagcaggtctgc ttggctctcatgctgggctcttacctggctggtctagtgagtttagtagcccacactaccctcaccttcagcctgagttactgtggttgctgc atatatttaaaggaaagagcaggacatøgaaaattaaattatagtatcttttt cctttttt^ tacatcttcattctcattgccatcctgaggatgcgttctgctgaaagtaggcgcaaagcgttctccacctgcgggtcccacctggtggca gtgactgtgttttatggaaccctgttctgcatgtacgttagacctcccacggacaggtcagtggaacagtccaaagtcattgctgttttcta cacttttgtaagccctatgttgaaccccatcatctatagtttgaggaacaaggatgtgaaacaagctttttggaaactgatcagaagaaac aaaagcatggcctgtggcagggtgggaaagacaaaatgttcagagaggccagagaaagacccatccatttgcagcgacagtgaaat tgtggctgctgttgtgaaggaa (SEQ ED NO: 47)

MOOSE00886 ctgl5296 1927198..1927675, 1927700..1928051, 1949720..1949831

NSEVQRVLFVWLLIYWTVCGNMLIWΗTSSPTLASPVΥFFLANLSFEDTFYS SSMAPKLIADSLYEGRTISYECCMAQLFGAHFLGGVEΠLLTVMAYDRYVAICKPLHN TTEVITRHLCAMLVGVAWLGGFLHSLVQLLLVLWLPFCGPNVINHFAFACTNTYVIG LLVVANSGLICLLNFLMLAASYTVILYSLRSHSADGRCKALSTCGAHFIVVALFFVPCI FTYVHPFSTLPIDKNMALFYGILTPMLNPLIYTLRNEEVKNAMRKLFTWQANIQIQEI QRMPQRYSSRRATPPJHNVRFTKVEMKE (SEQ ED NO: 50) aactcagaggtacagagagttctctttgtggtctttttgctgatctatgtggtcacggtttgtggcaacatgctcattgtggtcact atcacctccagccccacgctggcttcccctgtgtattttttcctggccaacclatcctttattgacaccttttattcttcttctatggctcctaaa ctcattgctgactcattgtatgaggggagaaccatctcttatgagtgctgcatggctcagctctttggagctcattttttgggaggtgttgag atcattctgctcacagtgatggcttatgaccgctatgtggccatctgtaagcccctgcacaatactaccatcatgaccaggcatctctgtg ccatgcttgtaggggtggcttggcttgggggcttcctgcattcattggttcagctcctcctggtcctttggttgcccttctgtgggcccaatg tgatcaatcactttgcctttgcctgcaccaatacgtatgtcattggtctgctggtggttgccaacagtggtttaatctgcctgttgaacttcct catgctggctgcctcctacattgtcatcctgtactccttgaggtcccacagtgcagatgggagatgcaaagccctctccacctgtggagc ccacttcattgttgttgccttgttctttgtgccctgtatatttacttatgtgcata ggtøttctgacacctatgttgaatccactcatttataccctgagaaatgaagaggtaaaaaatgccatgagaaagctctttacatggcagg ccaacattcagattcaggaaatacagagaatgccacaaagatactcctcaagaagagcaactccaagacacataaatgtcagattcac caaagttgaaatgaaggaa (SEQ ED NO: 49)

MOOSE00899 ctgl7659 60714..60781, 124639..124720, 298068..298255,

298292..298925

DMESSFSFGVILAVLASLΠATNTLVAVAVLLLIHKNDGVSLCFTLNLAVADTL IGVAISGLLTDQLSSPSRPTQKTLCSLP_ _\FVTSSAAASVLTVMLITFDRYLAIKQPFR YLKIMSGFVAGACIAGLWLVSYLIGFLPLGIPMFQQTAYKGQCSFFAVFHPHFVLTLS CVGFFPAMLLF FYCDMLKIASMHSQQIRKMEHAGAMAGALRTVSVLIGSFALSW TPFLITGIVQVACQECHLYLVLERYLWLLGVGNSLLNPLIYAYWQKEFRNILLFCLTC VSITSLRKKLFRC_ΪHVANFICIHMHTHTHTHTHTHTHT (SEQ ED NO: 52) gacatggaatcatctttctcatttggagtgatccttgctgtcctggcctccctcatcattgctactaacacactagtggctgtggc tgtgctgctgttgatccacaagaatgatggtgtcagtctctgcttcaccttgaatctggctgtggctgacaccttgattggtgtggccatctc tggcctactcacagaccagctctccagcccttctcggcccacacagaagaccctgtgcagcctgcggatggcatttgtcacttcctccg cagctgcctctgtcctcacggtcatgctgatcacctttgacaggtaccttgccatcaagcagcccttccgctacttgaagatcatgagtgg gttcgtggccggggcctgcattgccgggctgtggttagtgtcttacctcattggcttcctcccactcggaatccccatgttccagcagact gcctacaaagggcagtgcagcttctttgctgtatttcaccctcacttcgtgctgaccctctcctgcgttggcttcttcccagccatgctcctc tttgtcttcttctactgcgacatgctcaagattgcctccatgcacagccagcagattcgaaagatggaacatgcaggagccatggctgga gctctccgtactgtgtctgttctcattgggagctttgctctatcctggacccccttccttatcactggcattgtgcaggtggcctgccagga gtgtcacctctacctagtgctggaacggtacctgtggctgctcggcgtgggcaactccctgctcaacccactcatctatgcctattggca gaaggagttccgtaacattctgctcttctgcctcacatgtgtctctattacctctttgagaaagaagctctttcgttgtattcaccatgtggcta actttatatgtatacatatgcatacacacacacacacacacacacacacacacacacacaca (SEQ ED NO: 51)

MOOSE00930 ctgl8867 3972483..3972607, 4167256..4168117

SQDWRTΠPALLVAVCLVGFVGNLCVIGILLHNAWKGKPSMIHSLILNLSLAD LSLLLFSAPIRATAYSKSVWDLGWFVCKSSDWFIHTCMAAKSLΉVWAKVCFMYA SDPAKQVSIHNYTIWSVXVAIWTVASLLPLPEWFFSTIRHHEGVEMCLVDVPAVAEE FMSMFGKLYPLLAFGLPLFFASFYFWP^YDQCKKRGTKTQNLRNQIRSKQVTVMLL SIAΠSALLWLPEWVAWLWVWHLKAAGPAPPQGFIALSQVLMFSISSANPLIFLVMSE EFREGLKDSILRIHFVCSSMRSSYSSVKVFLCDSSNSGTSSDSTSNARS (SEQ ED NO:

54) tcccaggactggagaaccatcatcccggctctcttggtggctgtctgcctggtgggcttcgtgggaaacctgtgtgtgattgg catcctccttcacaatgcttggaaaggaaagccatccatgatccactccctgattctgaatctcagcctggctgatctctccctcctgctgt tttctgcacctatccgagctacggcgtactccaaaagtgtttgggatctaggctggtttgtctgcaagtcctctgactggtttatccacacat gcatggcagccaagagcctgacaatcgttgtggtggccaaagtatgcttcatgtatgcaagtgacccagccaagcaagtgagtatcca caactacaccatctggtcagtgctggtggccatctggactgtggctagcctgttacccctgccggaatggttctttagcaccatcaggca tcatgaaggtgtggaaatgtgcctcgtggatgtaccagctgtggctgaagagtttatgtcgatgtttggtaagctctacccactcctggca tttggccttccattattltttgccagcttttatttctggagagcttatgaccaatgtaaaaaacgaggaactaagactcaaaatcttagaaacc agatacgctcaaagcaagtcacagtgatgctgctgagcattgccatcatctctgctctcttgtggctccccgaatgggtagcttggctgtg ggtatggcatctgaaggctgcaggcccggccccaccacaaggtttcatagccctgtctcaagtcttgatgttttccatctcttcagcaaat cctctcatttttcttgtgatgtcggaagagttcagggaaggcttgaaagattccatcttaagaatccactttgtttgctcatccatgagaagc agttactcatctgttaaagttttcttatgcgatagcagcaattcaggcacatcatcagactccacttctaatgccaggtct (SEQ ID NO: 53)

MOOSE00941 ctg3235 477077..477375, 493498..493721, 494218..494258,

519487..519562, 557358.-557534, 582821..582914, 612454..612496 CYKQTLSFTGLTCIVSLVALTGNAVVTWLLGCRMRRNAVSIYILNLVAADFL

FLSGffllCSPLP .IMRJfflSKILSPVMTFPYFIGLSMLSAISTERCLIKHCISALWPIWYH

CRRPTHLSAVLCALLWAPSLLLAFLEGYYCAFLFKIGDYSWFQTFDFITGTWLIFKFV VLFYVTLKIMFCSEIGITSVSHRAQPΉSLTHMVASYLGVMLLLFLICSLPLGIKWFLLF WILVDFDIFLCHLQPVSDVLSSLNSSANPIIYFFMGSFRQR CCCCCCCCCCFTKFLL GITFVSRRGGSHLHSGHSRHVPSHSRH (SEQ ED NO: 56) tgctacaagcagaccctgagcttcacggggctgacgtgcatcgtttcccttgtcgcgctgacaggaaacgcggttgtgctct ggctcctgggctgccgcatgcgcaggaacgctgtctccatctacatcctcaacctggtcgcggccgacttcctcttccttagcggccac atlatatgttcgccgttacgccteatcaatatccgccatcccatctccaaaatcctcagtcctgtgatgacctttccctactttataggcctaa gcatgctgagcgccatcagcaccgagcgctgcctcatcaaacactgcatatctgccctgtggcccatctggtaccactgccgtcgccc cacacacctgtcagcag cctgtgtgccctgctctgggccccgtccctgctgcttgccttcctggaaggttactactgtgcttttctgtttaa gattggggactacagttggtttcagacatttgatttcatcacaggcacgtggctgatttttaaatttgtggttctcttctatgttcccctgaaaa tcatg tctgttctgagattggaattacaagcgtgagccaccgcgcccagccgacaatctctttaacccatatggttgcatcatatctagga gtaatgttgctgctcttccttatttgcagcctgcccttaggcattaagtggttcctattøttctggatcctcgtggattttgatatcttcctttgtca tttgcaaccagtttcagatgtcctgtcctotcttaacagcagtgccaaccccatcatttacttcttcatgggctcctttaggcagcgtgtttttt gttgttgflgttgttgttgctgctgttgttttacaaaattcctgttaggaataacttttgtgtccaggcgcggtggctcacacctgcattccggc cattccaggcatgtcccctcccattccaggcat (SEQ ED NO: 55)

MOOSE00981 ctgl590742265065..42265689, 42265723..42265979,

42310781..42310870 ERLLRLISAGVCGLILLVGLSANGLMLLWGRGPGSPHPLHSLTHSLMMNITP

SDLLFLACWPVLLLSFLQHNWWLGPAICΗSQATNTATTFCIFYSMVATALLRHVA VARPDLAFPAGWGTLLLLCGAMWALGLTESLPNWLFQRVAVEEETAGAPKTQACL LLLSPAGTSCYISLLGALAFLPCTLGLGCSFSHVGWLLWTQPQENIGLSLWLWFVL MWGPCSMLGYVAAMGYLPATPAAFVASSLCTILAYSNCAVSPILCFYLSRPFQAGLR DLFCRPMMAP_IPRGPGQQERDSVSNNKTKTKTH1ΗTΗ'IΗ^,IΗTΗT (SEQ ED NO: 58) gagcgactgctccgactcatctctgctggggtctgtggcctcatcctgctggtggggctgtcagctaatgggctcatgctgct ggtggtgggccggggcccgggctccccccacccgctccactccctgacccacagcctcatgatgaacatcacgccatctgacctgct cttcctggcctgcgtggtgcctgtgctgctgctgagcttcctgcagcacaactggtggctgggccctgccatctgcaccattagccagg ccaccaacacagccaccacgttctgcatcttctatagcatggtggccacagctctcctgcgccatgtggctgtggcccggcctgacctg gccttcccagccggctggggcaccctcttgctgctctgtggggccatgtgggccctgggccttacagaatccctgcccaactggctgtt ccagagggtggcagtggaggaggagacagcgggggctcccaagacccaggcctgcctcttgctcctgagccctgctgggacctcc tgctacatcagcctgctgggagccctggccttcctgccatgcacgctggggctgggctgctctttcagccacgtgggctggctcctgtg gacccagccccaagagaacatagggctcagccttgtggtgctggtggtttttgtgctgatgtgggggccctgctccatgctgggttatgt ggcagccatgggctacctgcctgccacaccggctgcttttgtggcctccagcctctgcaccatcctggcctactccaattgcgctgtca gccctatcctctgcttctacctctcccgccccttccaggcaggactcagggacctcttctgcaggccgatgatggccaggcatcccaga ggtcctgggcaacaagagcgagactctgtctcaaataataaaacaaaaaccaaaacacacacacacacacacacacacacacacac acacaca (SEQ ED NO: 57) MOOSE00994 ctgl7802 159288..159335, 297487..298009, 298142..298330,

313005..313117

SLALYNVFPFFFWLLFVGALLGNGALLVVVLRTPGLRDALYLAHLCVVDLLA AASIMPLGLLAAPPPGLGRVRLGPAPCRAARFLSAALLPACTLGVAALGLARYRLIV HPLRPGSRPPPVLVLTAVWAAAGLLGALSLLGPPPAPPPAPARCSVLAGGLGPFRPL WALLAFALPALLLLGAYGGEFWAPLAVGQFAACWLPYGCACLAPAARAAEAEAA VTWVAYSAFAAHPFLYGLLQRPVRLALGRLSRLRSSWAVRTTSGSRLLRGPCSVLD QTLSSSGPSLAGSS (SEQ ED NO: 60) tctttøgcactctacaatgtctttccattttttttctggcttctttttgtgggggcactgctgggcaacggcgcgctgctggtcgtg gtgctgcgcacgccgggactgcgcgacgcgctctacctggcgcacctgtgcgtcgtggacctgctggcggccgcctccatcatgcc gctgggcctgctggccgcaccgccgcccgggctgggccgcgtgcgcctgggccccgcgccatgccgcgccgctcgcttcctctcc gccgctctgctgccggcctgcacgctcggggtggccgcacttggcctggcacgctaccgcctcatcgtgcacccgctgcggccagg ctcgcggccgccgcctgtgctcgtgctcaccgccgtgtgggccgcggcgggactgctgggcgcgctctccctgctcggcccgccg cccgcaccgccccctgctcctgctcgctgctcggtcctggctgggggcctcgggcccttccggccgctctgggccctgctggccttc gcgctgcccgccctcctgctgctcggcgcctacggcggcatcttcgtggtggcgccgctggccgtgggccaatttgcagcctgctgg ctgccttatggctgcgcgtgcctggcgcccgcagcgcgggccgcggaagccgaagcggctgtcacctgggtcgcctactcggcctt cgcggctcaccccttcctgtacgggctgctgcagcgccccgtgcgcttggcactgggccgcctctctcgcctgaggagcagctgggc ggtgaggaccacatctggatcaagactgctcagaggcccttgcagtgtgcttgaccaaacgctgtcaagttcagggcccagcctggct gggtcctcc (SEQ ED NO: 59)

MOOSE01139 ctgl26341689941..1690075, 1696196..1696364, 1729541..1729632, 1731449..1731515, 1744203..1744423, 1748034..1748243, 1777967..1778134, 1813481..1813504, 1865325..1865428, 1868201..1868336

THLPSASSQIPALEESCEAVEAREIMWFKTRQGQIAKQPCPAGΉGVSTYLCLA PDGIWDPQGPDLSNCSSPWVNHITQKRSCRAYVQSEENFNPNCSFWSYSKRTMTGY

WSTQGCRLLTTNKTHTTCSCNHLTNFAVLMAHVEHSDAVHDLLLDVITWVGILLSL VCLLICIFTFCFFRGLQSDRNTIHKNLCISLFVAELLFLIGINRTDQPIACAVFAALLHFF FLAAFTWMFLEGVQLYIMLVΈVFESEHSRRKYFYLVGYGMPALIVAVSAAVDYRSY GTDKVCWLRLDTYFIWSFIGPATLIIMLNVΠ^LGIALYKMFHHTAILKPESGCLDNIKS WVIGAIALLCLLGLTWAFGLMYINESTVTMAYLFTIJFNSLQGMFIFIFHCVLQKKVRK EYGKCLRTHCCSGKSTESSIGSGKTSGSRTPGRYSTGSQVNN (SEQ ED NO: 62) acacaccttccatcagcatcgtcccaaatcccagctctcgaagagagctgtgaggctgtggaagcccgagaaatcatgtgg tttaagactcgtcaaggacagatagcaaagcagccatgccctgcaggaactataggtgtatcaacttatctatgccttgctcctgatgga atttgggatccccaaggtccagatctcagcaactgttcttctccttgggtcaatcatataacacagaagcgctcttgcagagcctatgtcc agtcagaggaaaatttcaaccctaactgttcattttggagctactccaagcgtacaatgacaggttattggtcaacacaaggctgtcggct cctgacaacaaataagacacatactacatgctcttgtaaccacctaacaaattttgcagtactgatggcacatgtggaacacagtgatgc ggtccatgacctccttetggatgtgatcacgtgggttggaattttgctgtcccttgtttgtctcctgatttgcatcttcacattttgctttttccgg gggctccagagtgaccgtaacaccatccacaagaacctctgcatcagtctctttgtagcagagctgctcttcctgattgggatcaaccga actgaccaaccaattgcctgtgctg tttcgctgccctgttacatttcttcttcttggctgccttcacctggatgttcctggagggggtgcag ctttatatcatgctggtggagg ttttgagagtgaacattcacgtaggaaatacttttatctgg cggctatgggatgcctgcactcattgtg gctgtgtcagctgcagtagactacaggagttatggaacagataaag atgttggctccgacttgacacctacttcatttggagttttatagg accagcaactttgataattatgcttaatgtaatcttccttgggattgctttatataaaatgtttcatcatactgctatactgaaacctgaatcagg ctgtcttgataacatcaagtcatgggttataggtgcaatagctcttctctgcctattaggattgacctgggcctttggactcatgtatattaat gaaagcacagtcatcatggcctatctcttcaccattttcaattctctacagggaatgtttatatttattttccattgtgtcctacagaagaaggt acgaaaagagtatgggaaatgcctgcgaacacattgctgtagtggcaaaagtacagagagttccattggttcagggaaaacatctggtt ctcgaactcctggacgctactccacaggctcacaggtaaacaat (SEQ ED NO: 61)

MOOSE01146 ctgl5998 3076899..3076950, 3227362..3227381, 3254354..3254409,

3255136..3255184, 3257523..3257602, 3334521..3334629, 3354815..3354918, 3355747.-3355867, 3358761..3358914, 3368419..3368485, 3370880..3370950, 3371577..3371604, 3415043..3415096, 3441342..3441468, 3442278.-3442323, 3512965..3513101

HRRLKQENHLNPGGRGCMGLWGGGLCWVPTAPGSCGAPPGRLRCPDWFSA LSSTSMMSGPSLSLCCPHGPRCLQTGHWSPRMQATVSGEGVWSNHGCALTRGNLTY SVCRCTHLTNFAILMQLARGHQVALSSISYVGCSLSVLCLVATLVTFAVLSTIRNQRY HIHANLSFAVLVAQVLLLISFRLEPGTVSGRSSTPCQVMAVLLHYFFLSAFAWMLVE GLHLYSMVIKVFGSEDSKHRYYYGMGWGFPLLICπSLSFAMDSYGTSNNCWLSLAS GAIWAFVAPALFλ vNPNFVFLVETGPVLLTGLASQDPSVRSTQLTAKAAAVLLPILG TSWVFGVLAVNGCAWFQYMFATLNSLQGLFIFLFHCLLNSEVQGLEELQKKWWG GDPELGISRKPLPSGAAGAGKGEHSQGLGGKSRA (SEQ ED NO: 64) cacaggaggctgaagcaggagaatcacttgaacccgggaggcagaggttgcatggggctttggggtgggggtctttgct gggtgcccaccgcgccagggagctgcggtgctcccccgggccgcctgcggtgccccgactggttctctgccctgtcctcaactagc atgatgtcaggtccttcactctccctctgctgtcctcacggacccaggtgtctgcagacaggacactggagccccaggatgcaggcca ctgtctccggagaaggggtctggtcgaaccacggctgtgcgctcacgagaggaaacctcacctactccgtctgccgctgcactcacct caccaactttgccatcctcatgcagcttgcacgcggacaccaggtggcgctgtcgtctatcagctatgtgggctgctccctctccgtgct ctgcctggtggccacgctggtcaccttcgccgtgctgtccaccatccggaaccagcgctaccacatccacgccaacctgtccttcgcc gtgctggtggcccaggtcctgctgctcattagtttccgcctcgagccgggcacggtgagtgggcgcagctccaccccctgccaagtg atggccgtgctcctacactacttcttcctgagtgccttcgcatggatgctggtggaggggctgcacctctacagcatggtgatcaaggtc tttgggtcggaggacagcaagcaccgttactactatgggatgggatggggttttcctcttctgatctgcatcatttcactgtcatttgccatg gacagttacggaacaagcaacaattgctggctgtcgttggcgagtggcgccatctgggcctttgtagcccctgccctgtttgtcatcgtg gtacctaattttgtatttttagtagagacgggtccagttttgcttactggactagcgagccaagacccttctgtgcgctccacccagttgac ggccaaggcagcggccgtgctgctgcccatcctgggtacctcgtgggtctttggcgtgcttgctgtcaacggttgtgctgtggttttcca gtacatgtttgccacgctcaactccctgcagggactgttcatattcctctttcattgtctcctgaattcagaggtacaggggctggaggag ctgcagaagaaatggtggggcggtgaccctgagttaggcatcagcaggaagccactgccatccggggctgcaggggccgggaag ggtgagcacagccagggcctgggtggcaagtccagggca (SEQ ED NO: 63)

MOOSEOl 148 ctgl2559 20179213..20179257, 20217038..20217100,

20401216..20401250, 20420415..20420483, 20567869..20567971, 20571553..20571656, 20572335..20572624, 20573833..20573929, 20576381..20576664, 20593994..20594078, 20661185..20661284

YYQEQLAQKDPLTYLNDNCFILPDIFTCRFTCPWGQSCSLCPYLCPEWFQLFP SAQVIRSCTKWSGTWDTFFGGWNTSGCVAHRDSDASETVCLCNHFTHFGVLM LDAJ_NTKVXTFISYIGCGISAIFSAATLLTYVAFEFLFSFFRKLRRDYPSKILMNLSTAL LFLNLLFLLDGWITSI^NVDGLCIAVAVTLITFFLLATFTWMGLEAIHMYIALVKVFNT YIP IYILKFCΠGWGLPALVVSVVLASRNNNEVYGKESYGKEKGDEFCWIQDPVIFYV TCAGYTGVT_FFLNLAMFRVΥMVQICGRNGKRSNRTLREEVLRNLRSVVSLTFLLGMT WGFAFFAWGPLNIPFMYLFSIFNSLQGLFIFIFHCAMKENVQKQWRQHLCCGRFRGTI SAHCKLRLPGSRHSPASASQVAGTTGTSHH (SEQ ED NO: 66) tattatoaggaacaattggcacagaaagaccctttgacgtacttaaatgataattgctttattcttcctgatattttcacttgtagatt cacctgcccatggggccaatcttgtagtctttgcccatacctatgtcctgaatggttccagttgtttccttctgctcaggttatcagaagctg cacaaagtggtctggtacttgggacactttttttggaggatggaacacgtcaggatgtgttgcacacagagattcagatgcaagtgaga cagtctgcctgtg aaccacttcacacactttggagttctgatgttagatgcaagaaacactaaagtcctcactttcatcagctatattgggt gtggaatatctgctattltttcagcagcaactctcctgacatatgttgcttttgaattccttttttcattitttaggaaattgcgaagg ctccaaaatcttgatgaacctgagcacagccctgctgttcctgaatctcctcttcctcctagatggctggatcacctccttcaatgtggatg gactttgcattgctgttgcagtcctgttgcatttcttccttctggcaacctttacctggatggggctagaagcaattcacatgtacattgctct agttaaagtatttaacacttacattcgccgatacattctaaaattctgcatcattggctggggtttgcctgccttagtggtgtcagttgttctag cgagcagaaacaacaatgaagtctatggaaaagaaagt atgggaaagaaaaaggtgatgaattctgttggattcaagatccagtcata ttttatgtgacctgtgctgggtattttggagtcatgttttttctgaacattgccatgttcattgtggtaatggtgcagatctgtgggaggaatgg caagagaagcaaccggaccctgagagaagaagtgttaaggaacctgcgcagtgtggttagcttgacctttctgttgggcatgacatgg ggttttgcattctttgcctggggacccttaaatatccccttcatgtacctcttctccatcttcaattcattacaaggcttatttatattcatcttcca ctgtgctatgaaggagaatgttcagaaacagtggcggcagcatctctgctgtggtagatttcgtggcacgatctcggctcactgcaagc tccgcctccccggttcacgccattctcctgcctcagcctcccaagtagctgggactacaggcaccagccaccat (SEQ ED NO: 65)

MOOSE01165 ctgl5361 7292455.-7293335, 7295796..7295919, 7300668..7300880,

7306454..7307290, 7309398..7309701, 7328904..7328998

GHmGGLFAIHEKMLSSEDSPRRPQIQECVGFEISVFLQTLAMIHSIEMINNSTL LPGVKLGYEIYDTCTEVTVAMAATLRFLSKFNCSRETVEFKCDYSSYMPRVKAVIGS GYSEITMAVSRMLNLQLMPQVGYESTAEILSDKIRFPSFLRTVPSDFHQIKAMAHLIQ KSGW-^WIGIITTDDDYGRLALNTFΠQAEA_JNVCIAFKEVLPAFLSDNTIEVT_NRTLK KTILEAQV-TVTVVFLRQFHVFDLFNKAffiMNiNKMWIASDNWSTATKITΗPNVKΩ KWGFAFRRGNISSFHSFLQNLHLLPSDSHKLLHEYAMHLSACAYVKDTDLSQCIFN HSQRTLAYKANKAIERNFVMRNDFLWDYAEPGLIHSIQLAVFALGYAIRDLCQARD CQNPNAFQPWELLGVLKNVTFTDGWNSFHFDAHGDLNTGYDVNLWKEINGHMTV TKMAEYDLQ- TOVFΠPDQETKNEFRNLKQIQSKCSKECSPGQMKKTTRSQHICCYEC

QNCPENHYTNQTDMPHCLLCNNKTHWAPVRSTMCFEKEVEYLNWNDSLAILLLILS LLGΠFVLWGIIFTRNLNTPWKSSGGLRVCYVILLCHFLNFASTSFFIGEPQDFTCKT RQTMFGVSFTLCISCILTKSLKILLAFSFDPKLQKFLKCLYRPILIIFTCTGIQWICTLW LIFAAPTVEVNVSLPRVIILECEEGSILAFGTMLGYIAILAFICFIFAFKGKYENYNEAKF

ITFGMLIYFIAWITFIPIYATTFGKYVPAVΈI IUIS-ΓYGILYCTFIPKCYV^ SAFLK (SEQ ED NO: 68) ggacatatcataattggaggtttgtttgctattcatgaaaaaatgttgtcctcagaagactctcccagacgaccacaaatccag gagtgtgttggctttgaaatatcagtttttcttcaaactcttgccatgatacacagcattgagatgatcaacaattcaacactcttacctggag tcaaactggggtatgaaatctatgacacttgtacagaagtcacagtggcaatggcagccactctgaggtttctttctaaattcaactgctcc agagaaactgtggagtttaagtgtgactattccagctacatgccaagagttaaggctgtcataggttctgggtactcagaaataactatgg ctgtctccaggatgttgaatttacagctcatgccacaggtgggttatgaatcaactgcagaaatcctgagtgacaaaattcgctttccttca tttttacggactgtgcccagtgacttccatcaaattaaagcaatggctcacctgattcagaaatctggttggaactggattggcatcataac cacagatgatgactatggacgattggctcttaacacttttataattcaggctgaagcaaataacgtgtgcatagccttcaaagaggttcttc cagcctttctttcagataataccattgaagtcagaatcaatcggacactgaagaaaatcatt^ ttctgaggcaattccatgtttttgatctcttcaataaagccattgaaatgaatataaataagatgtggattgctagtgataattggtcaactgc caccaagattaccaccattcctaatgttaaaaagattggcaaagttgtagggtttgcctttagaagagggaatatatcctctttccattccttt cttcaaaatctgcacttgcttcccagtgacagtcacaaactcttacatgaatatgccatgcatttatctgcctgcgcatatgtcaaggacact gatttgagtcaatgcatattcaatcattctcaaaggactttggcctacaaggctaacaaggctatagaaaggaacttcgtcatgagaaatg acttcctctgggactatgctgagccaggactcattcatagtattcagcttgcagtgtttgcccttggttatgccattcgggatctgtgtcaag ctcgtgactgtcagaaccccaacgcctttcaaccatgggagttacttggtgtgctaaaaaatgtgacattcactgatggatggaattcattt cattttgatgctcacggggatttaaatactggatatgatgttgtgctctggaaggagatcaatggacacatgactgtcactaagatggcag aatatgacctacagaatgatgtcttcatcatcccagatcaggaaacaaaaaatgagttcaggaatcttaagcaaattcaatctaaatgctc caaggaatgcagtcctgggcaaatgaagaaaactacaagaagtcaacacatctgttgctatgaatgtcagaactgtcctgaaaatcatt acactaatcagacagatatgcctcactgccttttatgcaacaacaaaactcactgggcccctgttaggagcactatgtgctttgaaaagg aagtggaatatctcaactggaatgactccttggccatcctactcctgattctctccctactgggaatcatatttgttctggttgttggcataat atttacaagaaacctgaacacacctgttgtgaaatcatccgggggattaagagtctgctatgtgatccttctctgtcatttcctcaattttgcc agcacgagctttttcattggagaaccacaagacttcacatgtaaaaccaggcagacaatgtttggagtgagctttactctttgcatctcctg cattttgacgaagtctctgaaaattttgctagccttcagctttgatcccaaattacagaaatttctgaagtgcctctatagaccgatccttatta tcttcacttgcacgggcatccaggttgtcatttgcacactctggctaatctttgcagcacctactgtagaggtgaatgtctccttgcccaga gtcatcatcctggagtgtgaggagggatccatacttgcatttggcaccatgctgggctacattgccatcctggccttcatttgcttcatattt gctttcaaaggcaaatatgagaattacaatgaagccaaattcattacatttggcatgctcatttacttcatagcttggatcacattcatcccta tctatgctaccacatttggcaaatatgtaccagctgtggagattattgtcatattaatatctaactatggaatcctgtattgcacattcatcccc aaatgctatgttattatttgtaagcaagagattaacacaaagtctgcctttctcaag (SEQ ED NO: 67)

MOOSE01172 ctgl47791950842..1951724, 1953313..1953437, 1953760..1953972, 1954995..1955756, 1959173..1959470, 1967494..1967524, 1970244..1970307

GP VVGALFISΠ.ESWΎQEACPEIYCFHPPTCLGFNEHGYHLFQAMRLGVEEIN

NSTALLPNITLGYQLYDVCSDSANVYATLRVLSLPGQHHIELQGDLLHYSPTVLAVIG

PDSTNRAATTAALLSPFL MSYAASSETLSVKRQYPSFLRTIPNDKYQVΈTMVLLL

QKFGWTWISLVGSSDDYGQLGVQALENQATGQGICLAFKDIMPFSAQVGDERMQCL MRHLAQAGATVVVVFSSRQLARVFFESVVLTNLTGKVWVASEAWALSRHITGVPGI

QRIGMVLGVAIQKRAWGLKAFEEAYARADKKAPRPCHKGSWCSSNQLCRECQAF MAHTMPKLKAFSMSSAYNAYRAVYAVAHGLHQLLGCASGACSRGRVYPWQLLEQI

HKVHFLLHKDTVAFNDNRDPLSSYNΠAWDWNGPKWTFTVLGSSTWSPVQLNΓNET

KIQWHGKDNQVPKSVCSSDCLEGHQRWTGFHHCCFECVPCGAGTFLNKSGECQPC GKEEWAPEGSQTCFPRTWFLALREHTSWVLLAANTLLLLLLLGTAGLFAWHLDTP

VVRSAGGRLCFLMLGSLAAGSGSLYGFFGEPTRPACLLRQALFALGFTIFLSCLTVRS FQLIΠFKFSTK TFYHAWVQNHGAGLFVMISSAAQLLICLTWLVVWTPLPAREYQR FPHLVMLECTETNSLGFILAFLYNGLLSISAFACSYLGKDLPENYNEAKCVTFSLLFNF VSWIAFFTTASVYDGKYLPAANMMAGLSSLSSGFGGYFLPKCYVILCRPDLNSTEHF QA (SEQ ED NO: 70) ggcagactagtcgtaggagccctttttaatctøgaaagttgg accaagaagcatgtcctgagatttattgttttcatcctccca cctgcctaggcttcaatgagcatggctaccacctcttccaggctatgcggcttggggttgaggagataaacaactccacggccctgctg cccaacatcaccctggggtaccagctgtatgatgtgtgttctgactctgccaatgtgtatgccacgctgagagtgctctccctgccaggg caacaccacatagagctccaaggagaccttctccactattcccctacggtgctggcagtgattgggcctgacagcaccaaccgtgctg ccaccacagccgccctgctgagccctttcctggtgcccatgattagctatgcggccagcagcgagacgctcagcgtgaagcggcagt atccctctttcctgcgcaccatccccaatgacaagtaccaggtggagaccatggtgctgctgctgcagaagttcgggtggacctggatc tctctggttggcagcagtgacgactatgggcagctaggggtgcaggcactggagaaccaggccactggtcaggggatctgcattgct ttcaaggacatcatgcccttctctgcccaggtgggcgatgagaggatgcagtgcctcatgcgccacctggcccaggccggggccac cgtcgtggttgttttttccagccggcagttggccagggtgtttttcgagtccgtggtgctgaccaacctgactggcaaggtgtgggtcgc ctcagaagcctgggccctctccaggcacatcactggggtgcccgggatccagcgcattgggatggtgctgggcgtggccatccaga agagggctgtccctggcctgaaggcgtttgaagaagcctatgcccgggcagacaagaaggcccctaggccttgccacaagggctcc tggtgcagcagcaatcagctctgcagagaatgccaagctttcatggcacacacgatgcccaagctcaaagccttctccatgagttctgc ctacaacgcataccgggctgtgtatgcggtggcccatggcctccaccagctcctgggctgtgcctctggagcttgttccaggggccga gtctacccctggcagcttttggagcagatccacaaggtgcatttccttctacacaaggacactgtggcgtttaatgacaacagagatccc ctcagtagctataacataattgcctgggactggaatggacccaagtggaccttcacggtcctcggttcctccacatggtctccagttcag ctaaacataaatgagaccaaaatccagtggcacggaaaggacaaccaggtgcctaagtctgtgtgttccagcgactgtcttgaagggc accagcgagtggttacgggtttccatcactgctgctttgagtgtgtgccctgtggggctgggaccttcctcaacaagagtggtgaatgcc agccttgtgggaaagaagagtgggcacctgagggaagccagacctgcttcccgcgcactgtggtgtttttggctttgcgtgagcacac ctcttgggtgctgctggcagctaacacgctgctgctgctgctgctgcttgggactgctggcctgtttgcctggcacctagacacccctgt ggtgaggtcagcagggggccgcctgtgctttcttatgctgggctccctggcagcaggtagtggcagcctctatggcttctttggggaac ccacaaggcctgcgtgcttgctacgccaggccctctttgcccttggtttcaccatcttcctgtcctgcctgacagttcgctcattccaacta atcatcatcttcaagttttccaccaaggtacctacattctaccacgcctgggtccaaaaccacggtgctggcctgtttgtgatgatcagctc agcggcccagctgcttatctgtctaacttggctggtggtgtggaccccactgcctgctagggaataccagcgcttcccccatctggtgat gcttgagtgcacagagaccaactccctgggcttcatactggccttcctctacaatggcctcctctccatcagtgcctttgcctgcagctac ctgggtaaggacttgccagagaactacaacgaggccaaatgtgtcaccttcagcctgctcttcaacttcgtgtcctggatcgccttcttca ccacggccagcgtctacgacggcaagtacctgcctgcggccaacatgatggctgggctgagcagcctgagcagcggcttcggtgg gtattttctgcctaagtgctacgtgatcctctgccgcccagacctcaacagcacagagcacttccaggcc (SEQ ED NO: 68) MOOSE01176 ctgl7658 40622..41S14, 41641..41761, 41896..42099, 42214..43015,

49551..49850, 49952.-50046

GDYVLGGLFPLGEAEEAGLRSRTRPSSPVCTRFSSNGLLWALAMKMAVEEIN

NKSDLLPGLRLGYDLFDTCSEPVVAMKPSLMFLAKAGSRDIAAYCNYTQYQPRVLA VIGPHSSELAMVTGKFFSFFLMPQCLLALQVSYGASMELLSARETFPSFFRTVPSDRV

QLTAAAELLQEFGWNWVAALGSDDEYGRQGLSIFSALAAARGICIAHEGLVPLPRA

DDSRLGKVQDVLHQVNQSSVQWLLFASVHAAHALFNYSISSRLSPKVWVASEAWL

TSDL\ -VLGLPGMAQMGTVLGFLQRGAQLHEFPQYVKTHLALATDPAFCSALGEREQ

GLEEDWGQRCPQCDCITLQNVSAGLNHHQTFSVYAAVYSVAQALHNTLQCNASG CPAQDPVKPWQLLENMYNLTFHVGGLPLRFDSSGNVDMEYDLKLWVWQGSVPRL HDVGRFNGSLRTERLKIRWHTSDNQKPVSRCSRQCQEGQVRRVKGFHSCCYDCVDC EAGSYRQNPDDIACTFCGQDEWSPERSTRCFRRRSRFLAWGEPAVLLLLLLLSLALG LVLAALGLFVHHRDSPLVQASGGPLACFGLVCLGLVCLSVLLFPGQPSPARCLAQQP LSHLPLTGCLSTLFLQAAEIFVESELPLSWADRLSGCLRGPWAWLWLLAMLVEVAL CTWYLVAFPPEVVTDWHMLPTEALVHCRTRSWVSFGLAHATNATLAFLCFLGTFLV RSQPGCYNRARGLTFAMLAYFITWVSFVPLLANVQVVLRPAVQMGALLLCVLGILA AFHLPRCYLLMRQPGLNTPEFFLG (SEQ ED NO: 72) ggggactacgtgctgggggggctgttccccctgggcgaggccgaggaggctggcctccgcagccggacacggcccag cagccctgtgtgcaccaggttctcctcaaacggcctgctctgggcactggccatgaaaatggccgtggaggagatcaacaacaagtc ggatctgctgcccgggctgcgcctgggctacgacctctttgatacgtgctcggagcctgtggtggccatgaagcccagcctcatgttcc tggccaaggcaggcagccgcgacatcgccgcctactgcaactacacgcagtaccagccccgtgtgctggctgtcatcgggccccac tcgtcagagctcgccatggtcaccgggaagttcttcagcttcttcctcatgccccaatgcctcttggccttgcaggtcagctacggtgcta gcatggagctgctgagcgcccgggagaccttcccctccttcttccgcaccgtgcccagcgaccgtgtgcagctgacggccgccgcg gagctgctgcaggagttcggctggaactgggtggccgccctgggcagcgacgacgagtacggccggcagggcctgagcatcttct cggccctggccgcggcacgcggcatctgcatcgcgcacgagggcctggtgccgctgccccgtgccgatgactcgcggctgggga aggtgcaggacgtcctgcaccaggtgaaccagagcagcgtgcaggtggtgctgctgttcgcctccgtgcacgccgcccacgccctc ttcaactacagcatcagcagcaggctctcgcccaaggtgtgggtggccagcgaggcctggctgacctctgacctggtcatggggctg cccggcatggcccagatgggcacggtgcttggcttcctccagaggggtgcccagctgcacgagttcccccagtacgtgaagacgca cctggccctggccaccgacccggccttctgctctgccctgggcgagagggagcagggtctggaggaggacgtggtgggccagcg ctgcccgcagtgtgactgcatcacgctgcagaacgtgagcgcagggctaaatcaccaccagacgttctctgtctacgcagctgtgtat agcgtggcccaggccctgcacaacactcttcagtgcaacgcctcaggctgccccgcgcaggaccccgtgaagccctggcagctcct ggagaacatgtacaacctgaccttccacgtgggcgggctgccgctgcggttcgacagcagcggaaacgtggacatggagtacgac ctgaagctgtgggtgtggcagggctcagtgcccaggctccacgacgtgggcaggttcaacggcagcctcaggacagagcgcctga agatccgctggcacacgtctgacaaccagaagcccgtgtcccggtgctcgcggcagtgccaggagggccaggtgcgccgggtcaa ggggttccactcctgctgctacgactgtgtggactgcgaggcgggcagctaccggcaaaacccagacgacatcgcctgcaccttttgt ggccaggatgagtggtccccggagcgaagcacacgctgcttccgccgcaggtctcggttcctggcatggggcgagccggctgtgct gctgctgctcctgctgctgagcctggcgctgggccttgtgctggctgctttggggctgttcgttcaccatcgggacagcccactggttca ggcctcgggggggcccctggcctgctttggcctggtgtgcctgggcctggtctgcctcagcgtcctcctgttccctggccagcccagc cctgcccgatgcctggcccagcagcccttgtcccacctcccgctcacgggctgcctgagcacactcttcctgcaggcggccgagatc ttcgtggagtcagaactgcctctgagctgggcagaccggctgagtggctgcctgcgggggccctgggcctggctggtggtgctgctg gccatgctggtggaggtcgcactgtgcacctggtacctggtggccttcccgccggaggtggtgacggactggcacatgctgcccacg gaggcgctggtgcactgccgcacacgctcctgggtcagcttcggcctagcgcacgccaccaatgccacgctggcctttctctgcttcc tgggcactttcctggtgcggagccagccgggctgctacaaccgtgcccgtggcctcacctttgccatgctggcctacttcatcacctgg gtctcctttgtgcccctcctggccaatgtgcaggtggtcctcaggcccgccgtgcagatgggcgccctcctgctctgtgtcctgggcat cctggctgccttccacctgcccaggtgttacctgctcatgcggcagccagggctcaacacccccgagttcttcctggga (SEQ ED NO: 71)

MOOSE01371 ctgl3655 22484333-22486189

ALALLQRSSGAAAASAKELACQEITVPLCKGIGYNYTYMPNQFNHDTQDEAG LEVHQFWPLVEIQCSPDLKFFLCSMYTPICLEDYKKPLPPCRSVCERAKAGCAPLMR QYGFAWPDRMRCDRLPEQGNPDTLCMDYNRTDLTTAAPSPPRRLPPPPPGEQPPSGS GHGRPPGARPPHRGGGRGGGGGDAAAPPARGGGGGGKARPPGGGAAPCEPGCQCR APMVSVSSERHPLYNRVT TGQIANCALPCHNPFFSQDERAFTVFWIGLWSVLCFVST FATVSTFLIDMERFKYPERPIIFLSACYLFVSVGYLVRLVAGHEKVACSGGAPGAGGA GGAGGAAAGAGAAGAGAGGPGGRGEYEELGAVEQHVRYETTGPALCTVVFLLVYF FGMASSIWWVDLSLTWFLAAGMKWGNEAIAGYSQYFHLAAWLWSVKSIAVTALSS \ GDPVAGICYVGNQSLDNLRGFVTAPLVR_ LFIGTMFLLAGFVSLFRIRSVIKQQDG PTKTHKLEKLMIRLGLFTVLYTWAAVVVACLFYEQHNRPRWEATHNCPCLRDLQP DQARRPDYAVFMLKYFMCLVVGITSGVWVWSGKTLESWRSLCTRCCWASKGAAV GGGA (SEQ ED NO: 74) gccttggcgctgctgcagcgctctagcggcgctgcggccgcctcggccaaggagctggcatgccaagagatcaccgtg ccgctgtgtaagggcatcggctacaactacacctacatgcccaatcagttcaaccacgacacgcaagacgaggcgggcctggaggt gcaccagttctggccgctggtggagatccagtgctcgcccgatctcaagttcttcctgtgcagcatgtacacgcccatctgcctagagg actacaagaagccgctgccgccctgccgctcggtgtgcgagcgcgccaaggccggctgcgcgccgctcatgcgccagtacggctt cgcctggcccgaccgcatgcgctgcgaccggctgcccgagcaaggcaaccctgacacgctgtgcatggactacaaccgcaccgac ctaaccaccgccgcgcccagcccgccgcgccgcctgccgccgccgccgcccggcgagcagccgccttcgggcagcggccacg gccgcccgccgggggccaggcccccgcaccgcggcggcggcaggggcggtggcggcggggacgcggcggcgcccccagct cgcggcggcggcggtggcgggaaggcgcggccccctggcggcggcgcggctccctgcgagcccgggtgccagtgccgcgcg cctatggtgagcgtgtccagcgagcgccacccgctctacaaccgcgtcaagacaggccagatcgctaactgcgcgctgccctgcca caacccctttttcagccaggacgagcgcgccttcaccgtcttctggatcggcctgtggtcggtgctctgcttcgtgtccaccttcgccac cgtctccaccttccttatcgacatggagcgcttcaagtacccggagcggcccattatcttcctctcggcctgctacctcttcgtgtcggtg ggctacctagtgcgcctggtggcgggccacgagaaggtggcgtgcagcggtggcgcgccgggcgcggggggcgctgggggcg cgggcggcgcggcggcgggcgcgggcgcggcgggcgcgggcgcgggcggcccgggcgggcgcggcgagtacgaggagct gggcgcggtggagcagcacgtgcgctacgagaccaccggccccgcgctgtgcaccgtggtcttcttgctggtctacttcttcggcatg gccagctccatctggtgggtgatcttgtcgctcacatggttcctggcggccggtatgaagtggggcaacgaagccatcgccggctact cgcagtacttccacctggccgcgtggcttgtgcccagcgtcaagtccatcgcggtgctggcgctcagctcggtggacggcgacccg gtggcgggcatctgctacgtgggcaaccagagcctggacaacctgcgcggcttcgtgctggcgccgctggtcatctacctcttcatcg gcaccatgttcctgctggccggcttcgtgtccctcttccgcatccgctcggtcatcaagcaacaggacggccccaccaagacgcacaa gctggagaagctgatgatccgcctgggcctgttcaccgtgctctacaccgtgcccgccgcggtggtggtcgcctgcctcttctacgag cagcacaaccgcccgcgctgggaggccacgcacaactgcccgtgcctgcgggacctgcagcccgaccaggcacgcaggcccga ctacgccgtcttcatgctcaagtacttcatgtgcctagtggtgggcatcacctcgggcgtgtgggtctggtccggcaagacgctggagt cctggcgctccctgtgcacccgctgctgctgggccagcaagggcgccgcggtgggcgggggcgcg (SEQ ED NO: 73) MOOSE01451 ctgl7341 5903471..5903744, 5904206..5904356, 5910947..5911003,

5919010..5919127, 5934289.-5934313, 5939959..5940053, 5941313..5941394, 5947946..5948017, 5948441..5948532

LARSSNPFEEKRHSGFLNFQLFCSGFSPSLCCQTGTFRSAEVFYCLLFSCALLT DQKEKMALHFLFHQTLQLIHLELEKMILKNLFMPYWDVWHHQFQYANQILGKGLY CFFR YSCSFVΕLAVSRDRATALQPGRQSETLSQKKKKRNATDFCMLSLCIEGT YLQIIRAICC TATGP FTQCVTKGSFKAQWQETTYNLFTFCCLFLLPLTAMVICYS RIVLSVTFSKFLAPAGEFALPRSFDNCPRVRLRALRLALLILLTFILCWTPYYLLGMWY WFSPTMLTEVPPSLSHILFLLGLLNAPLDPLLYGAFTL (SEQ ED NO: 76) ctggcaaggagcagcaatccttttgaggagaagaggcactctggttttttgaattttcagcttttctgctctggtttctccccatct ttgtgctgtcagacagggacatttaggtctgcagaag tttctattgccttttgttcagctgtgccctgctcacagaccagaaggagaagat ggctttacattttcttttccatcagacacttcaactcatacatttggagctggaaaagatgattttaaaaaatcttttcatgccctattgggatgt gtggcatcatcagttteaatatgcaaatcaaattttgggtaagggtctatactgctt^ gcttgcagtgagccgagatcgtgccactgcactccagcctgggcgacagagcgagactctgtctcaaaaaaaaaaaaaaagaaatg caactgatttttgcatgttgtctttgtgtattgaaggaacatacctocaaataataagagccatttgttgcaaacccacagccactggccca gtccctttcactcagtgtgtcaccaaaggcagcttcaaggctcaatggcaagagaccacctataacctcttcaccttctgctgcctctttct gctgccactgactgccatggtcatctgctatagccgcattgtcctcagtgttatattttctaaattcctagcccctgctggtgaatttgccctc ccccgctcctttgacaattgtccccgtgttcgtctccgggccctgagactggccctgcttatcttgctgaccttcatcctctgctggacacc ttattacctactgggtatgtggtactggttctcccccaccatgctaactgaagtccctcccagcctgagccacatccttttcctcttgggcct cctcaatgctcctttggatcctctcctctatggggccttcaccctt (SEQ ID NO: 75)

MOOSE01609 ctg37223820..223885, 415276..416383, 418102..418139

LGAVTTPVIPALGDAEAGRSSEVTVLHYNYTGKLRGARYQPGAGLRADAVVC

LAVCAFIVLENLAVLLVLGRHPRFHAPMFLLLGSLTLSDLLAGAAYAANILLSGPLTL KLSPALWFAREGGVFVALTAS VLSLLAIALERSLTMARRGPAPVSSRGRTLAMAAA AWGVSLLLGLLPALGWNCLGRLDACSTVLPLYAKAYVLFCVLAFVGILAAICALYA RJΎCQVRANARRLPAP 'GTAGTTSTRARRKPRSLALLRTLSVVLLAFVACWGPLFLL LLLDVACPARTCPVLLQADPFLGLAMANSLLNPIIYTLTNRDLRHALLRLVCCGRHS CGRDPSGSQQSASAAEASGGLRRCLPPGLDGSFSGSERSSPQRDGLDTSGSTGSPACV MKSCLLGKHK (SEQ ED NO: 78) ctgggcgcagtgactacgcctgtaatcccagcacttggggacgccgaggcaggcaggtcatctgaggtcatcgtcctgca ttacaactacaccggcaagctccgcggtgcgcgctaccagccgggtgccggcctgcgcgccgacgccgtggtgtgcctggcggtgt gcgccttcatcgtgctagagaatctagccgtgttgttggtgctcggacgccacccgcgcttccacgctcccatgttcctgctcctgggca gcctcacgttgtcggatctgctggcaggcgccgcctacgccgccaacatcctactgtcggggccgctcacgctgaaactgtcccccg cgctctggttcgcacgggagggaggcgtcttcgtggcactcactgcgtccgtgctgagcctcctggccatcgcgctggagcgcagcc tcaccatggcgcgcagggggcccgcgcccgtctccagtcgggggcgcacgctggcgatggcagccgcggcctggggcgtgtcg ctgctcctcgggctcctgccagcgctgggctggaattgcctgggtcgcctggacgcttgctccactgtcttgccgctctacgccaaggc ctacgtgctcttctgcgtgctcgccttcgtgggcatcctggccgctatctgtgcactctacgcgcgcatctactgccaggtacgcgccaa cgcgcggcgcctgccggcacggcccgggactgcggggaccacctcgacccgggcgcgtcgcaagccgcgctcgctggccttgct gcgcacgctcagcgtggtgctcctggcctttgtggcatgttggggccccctcttcctgctgctgttgctcgacgtggcgtgcccggcgc gcacctgtcctgtactcctgcaggccgatcccttcctgggactggccatggccaactcacttctgaaccccatcatctacacgctcacca accgcgacctgcgccacgcgctcctgcgcctggtctgctgcggacgccactcctgcggcagagacccgagtggctcccagcagtc ggcgagcgcggctgaggcttccgggggcctgcgccgctgcctgcccccgggccttgatgggagcttcagcggctcggagcgctca tcgccccagcgcgacgggctggacaccagcggctccacaggcagccccgcctgtgtgatgaaatcatgtcttttaggaaaacataag (SEQ ED NO: 77)

MOOSE02359 ctgl50377933711..7933760, 7963115..7963181, 7973661..7973682, 7983319..7983352, 7998114..7998223, 7999445..7999609, 8000036..8000207, 8000817..8001020, 8001339..8001472, 8003204..8003358 SPQAPGTWAAAWV LPTΛ VPDHAHYTLGTVILLVGLTGMLGNLTVTYTFCR

SRSLRTPAMVFFLLNLAVSDFLMSFTQAPVFFTSSLYKQWLFGETGCEFYAFCGALFGIS SMITLTAIALDRYLVITRRLATFGVASKRRAAFVLLGVWLYALAWSLPPFFGWSAYV PEGLLTSCSWDYMSFTPAVRAYTMLLCCF FLPLLIΠYCYIFIFRAIRETGRALQTFG ACKGNGESLWQRQRLQSECKMAKIMLLVILLFVLSWAPYSAVALVAFAGYAHVLTP YMSS AVTAKAS FINPIΓ_ AITHPKYRSDAVASWQSRRLGVHLYPWSLVFCHPSEF ENHNWSTRDVAPGHLMTEMETRVLSILSQ (SEQ ED NO: 80) tctccgcaggcacctgggacttgggctgctgcctgggtccccctccccacggttgatgttccagaccatgcccactataccc tgggcacagtgatcttgctggtgggactcacggggatgctgggcaacctgacggtcatctataccttctgcaggagcagaagcctccg gacacctgccaacatgttcattatcaacctcgcggtcagcgacttcctcatgtccttcacccaggcccctgtcttcttcaccagtagcctct ataagcagtggctctttggggagacaggctgcgagttctatgccttctgtggagctctctttggcatttcctccatgatcaccctgacggc catcgccctggaccgctacctggtaatcacacgcccgctggccacctttggtgtggcgtccaagaggcgtgcggcatttgtcctgctg ggcgtttggctctatgccctggcctggagtctgccacccttcttcggctggagcgcctacgtgcccgaggggttgctgacatcctgctc ctgggactacatgagcttcacgccggccgtgcgtgcctacaccatgcttctctgctgcttcgtgttcttcctccctctgcttatcatcatcta ctgctacatcttcatcttcagggccatccgggagacaggacgggctctccagaccttcggggcctgcaagggcaatggcgagtccct gtggcagcggcagcggctgcagagcgagtgcaagatggccaagatcatgctgctggtcatcctcctcttcgtgctctcctgggctccc tattccgctgtggccctggtggcctttgctgggtacgcacacgtcctgacaccctacatgagctcggtgccagccgtcatcgccaaggc ctctgcaatccacaaccccatcatttacgccatcacccaccccaagtacaggtcagatgctgtggcttcctggcagtccagacgtcttgg ggtacacttgtacccctggtccctggtgttctgccaccctagcgagtttgaaaaccacaactggagcaccagggatgttgccccaggc cacctgatgacagaaatggagactagagtgttgtctatactaagccaa (SEQ ED NO: 79)

MOOSE02360 ctgl5907 53482006..53482053, 53509199..53509251,

53519616..53519786, 53524954..53525124, 53528381..53528715, 53541306..53541470, 53636121..53636147, 53655914..53655948, 53805160..53805178, 53813699..53813772, 53992100..53992118

SLLKIQKISRTWWRVPSTDVSCLPMWGDFRAVSSELEILVLTIGIFIFFLVGILS TFGNGYVXYMSSRRKKKLRPAEMTINLAVCDLGISVVGKPFTΠSCFCHRWVFGWIG CRWYGWAGFFFGCGSLITMTAVSLDRYLKICYLSYGVWLKRKHAYICLAAIWAY

ASFWTTMPLVGLGDYVPEPFGTSCTLDWWLAQASVGGQVFILNILFFCLLLPTAVIV FSYVKTIAKVKSSSKEVAHFDSRΓHSSHVLEMKLTKVAMLICAGFLIAWIPYAVVSVW SAFGRPDSIPIQLSWPTLLAKSAAMYNPΠYQQNFSISNFRDSLAQSSVPWIQCCYYP

QESRGQLERWESRDFVRMSAVSADLQKFQRN (SEQ ED NO: 82) tctctactaaaaatacaaaaaattagtcggacgtggtggcgggtgcctagcacagatgtgagctgcttacctatgtggggag acttcagggctg gagtagtgaactcgaaatcttagttttaacgattggaatttttatttttttccttgtagggattctgtccacan^ gatatgtcctttacatgtcttctagacgaaagaagaagctgagacccgctgaaataatgactatcaatttagcagtctgtgatctggggatt tcagttgtaggcaagccgttcaccatcatctcttgcttttgtcaccgctgggtgtttggctggatcggctgccgctggtatggatgggctg gatttttctttggctgtggaagccttatcaccatgactgctgtcagcctggatcgatatttgaaaatctgctatttatcttatggggtttggctg aaaagaaagcacgcctacatctgcctggcagccatctgggcctatgcttccttctggaccaccatgcccttggtaggtctgggggacta cgtacctgagcccttcggaacctcgtgcaccctggactggtggctggcccaggcctcggtagggggccaggttttcatcctgaacatc ctcttcttctgcctcttgctcccaacggctgtgatcgtgttctcctacgtaaagatcattgccaaggttaagtcctcttccaaagaagtagct catttcgacagtcggatccatagcagccatgtgctggaaatgaaactgacaaaggtagcgatgttgatttgtgctggattcctgattgcct ggattccttatgcagtggtgtctgtgtggtcagcttttggaaggccagactccattcccatacagctcctgtggtgccaaccctacttgca aaatctgcagcgatgtacaatcccatcatttaccaacaaaacttctcaatatcaaattttcgtgattccctagctcagagctcagtcccttgg attcagtgctgttattatcctcaagagagcaggggacagctagaaagggttgtagaatcgagagattttgtgaggatgtcagcagttagc gcagatctacaaaaattccagaggaat (SEQ ED NO: 81)

Table II

MOOSE02352 ctgl23762508254..2508274, 2512637..2512697, 2596061..2596100, 2609217..2609251, 2697256..2697388, 2697901..2698120, 2702134..2702373, 2703765..2703914, 2704936..2705391, 2785876..2785900, 2796650..2796801

MSVVLQDTQTLTIDSNINSLLIVALLVTGITGMHHHARLILKLFVSKSSINFAD

FISQSSSASPGGVDYILHGSTVTFQHGQNLSFINISIΓDDNESEFEEPIEILLTGATGGAV LGPJILVSRΠIAKSDSPFGVTRFLNQSKISIA-Π'NSTMILSLVLERTGGLLGEIQVNΓVVΈT VGPNSQEALLPQNRDIADPVSGLFYFGEGEGGVRΉILTIYPHEEIEVEETFIIKLHLVK GEAKUDSRAKDVTLTIQEFGDPNGVVQFAPETLSKKTYSEPLALEGPLLITFFVRRVK GTFGEIMVYWELSSEFDITEDFLSTSGFFTIADGESEASFDVHLLPDEVPEIEEDYVIQL VSVEGGAELDLEKSITWFSVYANDDPHGVFALYSDRQSILIGQNLIRSIQINITRLAGT FGDVAVGLRISSDHKEQPIVTENAERQLWKDGATYKVDVFGTFYYSFISCTDGAVN LTYIQΠSKPIFSHYTDAYIFLHYSLLNTFLCDNSHFKISRK (SEQ ED NO: 84) atgagtgttgttctgcaggatactcagacgttaacaatagactcaaacattaacagcctgctcatagttgccctgctggtaact gggattacaggcatgcaccaccacgcccggctaattttaaaattatttgtaagtaaaagtagcataaactttgctgatttcatctctcagag ctcctctgccagtcccggaggtgttgattacattttgcatggcagtacagtcacctttcagcatgggcaaaacttaagttttataaatatctc catcattgatgacaatgaaagtgaatttgaggagcccattgaaattctactcactggagctactggaggagcggtccttgggcgccacc tagtgagcagaatcataatagctaagagtgactctccctttggagttataaggtttctcaatcaaagcaaaatttctattgctaatcccaattc cacaatgattttatcactggtgctggagcggactggaggactcttgggagagattcaggtgaactgggagacagtaggacccaactct caagaagccttactgccacagaatagagacattgcagacccagtgagcgggttgttctattttggagaaggagaaggaggagtgaga accataattctgacaatctatcctcatgaagaaattgaagttgaagagacattcattattaaacttcatcttgtgaaaggagaagctaaatta gactccagagctaaagatgttacattaaccatacaagagtttggtgacccaaatggagttgttcagtttgctcctgaaactttgtctaagaa gacttattcagagcctctggctctggaagggcccctgctcattaccttctttgtcagaagagtcaagggcacctttggagagattatggttt actgggaattaagtagtgagtttgacattactgaagactttctttccaccagtggatttttcaccattgctgatggagagagtgaagctagct ttgatgttcatttgctaccagatgaggtacctgagatagaggaagattatgtgatccagcttgtttctgtagagggaggagccgaactgg atctggagaagagtatcacatggttctctgtttatgcaaatgatgacccacatggagtatttgccctgtattcggatcgccagtcaatactta ttgggcagaaccttattagatccatccaaattaacataacccggcttgctggaacatttggagatgtggctgttgggcttcgaatatcatcg gatcataaagaacagccgattgttaccgaaaatgcagagaggcagctggtggtcaaagatggtgccacatataaagtggacgtgtttg gaaccttttattacagcttcatctcctgtactgatggagctgtaaatctaacatacatccagatcatttcaaagcctattttttctcactacaca gatgcctatatøtttcttcattacagcctttlaaatacttttctctgtgataattcccactttaaaatttcgagaaag (SEQ ED NO: 83) MOOSE01169 ctgl3067 1981228..1981248, 1981792..1982139, 1984400..1984592,

1984626..1984712, 1985489..1985522, 1985564..1985647, 1985892..1985938, 1986646..1986768, 1988270..1988412, 1988754..1988824, 1989127-1989251, 1991650-1992536

GGGILFMLLFPSKIHLITYKVYFFFP_ _^QRERWMKAMFFL KEINKRKDILPN ITLGYQIFDTCFΉSKSVEAVLVFLTGQEENRPNFRNSTGAFPAGIVGAGGSFLSVPAS

RILGLYYLPQVGYTSTCVTLSDKYQFPSYLRV1ASDKIQSKAVVKRIQHFHFLTLSPRL

ECSGAILAHGNLCLPVAGITGVCHHARLIFVFLVETGFCHVAQADGVSLCCHAGVYN

SPASAPLVAGTTGAHHHAQLIFWLRYVTLLSLQKGQSCPNWMHYLGEEYFQHREQ HLLNPEARVAGTLEEQKRSQCGWKDLSIVYTYFCNVMYHNLAQRLVIFSMLFNSDL LWKTQHMKILISKINIKGKYFLGFQDDSWNHRSFTSRNRPLPHSVCTDVCPPGTRKGI RSEGEPICCFDSPCADGHVSRKPGERECEQCGEDYWSNAQKSECVLKEVEYLAYDE ALGFTLVILSVFGAFV T.AVTAVΎVIHPJITPLV-^ASDWQLGFLIQVSLΠMLLSSMLFI D HNWSCMAGQVTLALGFSLCLSCLLGKTSSLFLAYRISKSKTQLTSMHPLYRKIIV LISVLAEIGICTAYLILEPPMVYKNMESQNTKΠLGCNEISIEFLYSMFGIDAFLALLCFL TTFVARQLPDNYYΈGKCITFGML FIΓWMSFWVYLSTKGKFKMAVEIFAILASSHG

LLGCIFAPKCLΠLLRPERNTSEI VCG (SEQ ED NO: 86) gggggaggfattttatttatgctgcttttccccteaaagatccacctaatcacc gagattccgctggatgaaagccatgatccacatgatcaaggagattaataagaggaaggatattttgcccaacatcactctgggctatc agatctttgatacctgttttaccatctccaaatcagtggaagcagtcttggtatttcttacagggcaggaagaaaacaggcccaattttaga aacagcactggagcatttccggcaggaattgttggagcaggtggatcattcttatcagttcctgcttcaagaattctagggttatattatttg cctcaggtgggctatacctctacctgcgtgattcttagtgacaaataccagtttccatcttatcttcgtgtaatagccagcgataagatcca gtcgaaggctgtggtaaaacgtatccaacactttcactttctcactctgtcgcccaggctggagtgcagtggcgccatcctggctcatgg caacctctgcctcccagtagctgggattacaggtgtgtgccaccatgccagactaatttttgtatttttagtagagacagggttttgccatgt tgcccaggctgACGGAGTCTCGCTCTGTTGCCATGCTGGAGTATAcaattctcctgcctcagcccccct agtagctgggactacaggcgcacaccaccacgctcagctaatttttgtatttttaagatatgtaacacttctgtcattgcagaaaggacag agctgccctaatgtgttcatgcattatctaggggaagaatatttccagcacagggaacagcaccttctaaaccctgaggcgcgtgtggct ggcactttggaggaacagaaaagaag caatgtggctggaaagacttatctattgtgtatacatacttctgtaatgttatgtaccataacct agctcagagattggtaatamtctatgttarttaattcagatttøctctggaaaactcaacatøtgaaaattctgatttctaaaataaatataaa gggaaaatattttttaggctttcaagatgattcttggaatcataggtccttcacaagcaggaacagacctcttccccattcagtgtgtactga tgtgtgtcctcctgggactcggaaggggattcgttcagagggagaaccaatatgctgctttgactccatcccatgtgctgatggacacgt gtcacggaaaccaggtgaaagggagtgtgaacaatgtggtgaagactattggtcaaatgcacaaaagagcgagtgtgtgctgaaaga ggtggaataccttgcttatgatgaggccctgggattcacacttgtcattctttctgtctttggggcatttgtggtcttggcagtcacagctgtg tatgtgatacacaggcacactcccctggtgaacgccagtgactggcagctgggctttctcattcaggtttctctgatcatcatgctgctgtc gtccatgcttttcattgacaagccacacaactggtcctgcatggctggccaggtcactctggcactgggcttttctctttgcctgtcttgcct tcttggaaagactagttcactgtttttagcclacagaatttocaaatccaaaactcaacttacatccatgcaccccctttatcggaaaatcatt gtgctaatctctgttctagcggagattggcatatgtacagcctacttgatattggaacctcccatggtatacaagaacatggaatctcaaaa tacaaagatcattctgggatgcaatgaaatttccatagagtitttgtactcgatgtttggaatt aacttttgtggctcgccagttaccagataaltactatgaaggaaaatgcatcacctttgggatgcttgtcttttteatcatttggatgtcttttgt ccctg ttatttgagcaccaaaggcaagttcaaaatggctgtggaaatatttgcaatcttggcatccagccatggcttgttgggttgtatatt tgctcctaagtgcctcattattttgctgaggccagagaggaacaccagtgaaattgtttgtgga (SEQ ED NO: 85)

MOOSE00810 ctgl3284 7238060..7238539, 7247370..7247453, 7258514-7258731,

7329591-7329682, 7438551-7438609 RLVXAAVETTVLVLIFAVSLLGNVCALVLVARRRRRGATACLVLNLFCADLL

FISAFFLVLAVRWTEAWLLGPVACHLLFYVT^TLSGSVTILTLAAVSLERMVCIVHLQ RGVRGPGRRARAVLLALIWGYSAVAALPLCVFFRVVPQRLPGADQVSAPLCISWDV SFVTLNFLVPGLVWISYSKILQIRVSQQDFRLFRTLFLLMVSFFIMWSPΠIΉLLILIQNF KQDLVIWPSLFFW\^AFTFANSALNPILYNMTLRVFRHRVLEIRDKLWPLΉLHKFSS GTSLPLGHTHTQAHTHTHAHTHLHT (SEQ ED NO: 88) cggctggtgctggccgcggtggagacaaccgtgctggtgctcatctttgcagtgtcgctgctgggcaacgtgtgcgccctg gtgctggtggcgcgccgacgacgccgcggcgcgactgcctgcctggtactcaacctcttctgcgcggacctgctcttcatcagcgcta tccctctggtgctggccgtgcgctggactgaggcctggctgctgggccccgttgcctgccacctgctcttctacgtgatgaccctgagc ggcagcgtcaccatcctcacgctggccgcggtcagcctggagcgcatggtgtgcatcgtgcacctgcagcgcggcgtgcggggtc ctgggcggcgggcgcgggcagtgctgctggcgctcatctggggctattcggcggtcgccgctctgcctctctgcgtcttcttccgagt cgtcccgcaacggctccccggcgccgaccaggtgagcgcccctctgtgtatctcgtgggatgtctcttttgttactttgaacttcttggtg ccaggactgg cattgtgatcagttactecaaaattttacagatccgcgtgtcccagcaggacttccggctcttccgcaccctcttcctcct catggtctccttcttcatcatgtggagccccatcatcatcaccatcctcctcatcctgatccagaacttcaagcaagacctggtcatctggc cgtccctcttcttctgggtggtggccttcacatttgctaattcagccctaaaccccatcctctacaacatgacactcagagtcttcaggcac agggtcctggagattagggacaaactttggcccctgacaattttgcataagttcagctctggcacaagtctaccattgggacatacacac acacaggcacacacacacacacatgcacacacacacttgcacaca (SEQ ED NO: 87)

MOOSE01192 ctgl4473 2193277..2193464, 2196762..2196915, 2197105-2197234,

2208484-2208612, 2213869..2213892, 2217011..2217047, 2219508..2219537, 2222803..2222949, 2223833-2223862, 2224187-2224215, 2281784..2281826, 2317525-2317642

MGSGISSESKESAKRSKELEKKLQEDAERDARTVKLLLLGAGESGK STIVKQMKRΠQGKHYTSEESAAWSLSAWSNIHKSINLIHHINRTKNKNHMISSIEAEK AFHKIQQPLMLKTLNKQEREΉSNLWSGVNIQNLYSYTEYLIPACTTSYLNDLDRITA SGYWNEQDVLHSRVKTTGIIETQFSFKDLHFRMFDVGGQRSERKKWIHCFEGVTCII

FCAALSAYDMVLVEDEEVNRMHESLHLFNSICNHKWSTTSIVLFLNKKDIFQEKVT K\ -H.SICFPEYTGPNTFEDAGNYIKNQFLDLNLKKEDKEIYSHMTCATDTQNVKFVF DAVTDΠIKENLKDCGLF (SEQ ED NO: 90) atgggaagtggaattagttcagagagcaaggagtcagccaaaagatcaaaagaactggagaaaaagcttcaggaggatg ctgagcgagatgcaagaaccgtaaagctgctactattaggagcaggagaatctgggaaaagtactattgttaaacaaatgaagagaat aatacagggaaaacattatacatcggaggaatctgctgcatggagcttgagtgcctggtcgaacatacacaaatcaataaacttaatcc atcacataaacagaaccaagaacaaaaaccacatgattagctcaatagaagcagaaaaggccttccacaaaattcaacagcctttgat gctaaaaactctcaataaacaagagagagagacgataagtaatctatggtcaggggttaatatccaaaatttatattcatacaccgaatat Itaataccagcctgcacaacaagctaccttaatgatttagatagaataacagcatctgggtatgtgccaaatgaacaagatgttctccattc tcgagtgaaaacgactggaatcattgaaactcaattctcctttaaagacttgcacttcaggatgtttgatgtaggtggacagagatctgag agaaagaagtggattcactgctttgaaggagttacatgcattatattttgtgctgcacttagtgcctatgacatggtcctcgtggaagacga agaagtgaatagaatgcatgaaagccttcacctgttcaacagtatctgtaatcacaagtatttttcaacaacctccattgtcctgttcctcaa caaaaaagatøtctttcaagaaaaggtaaccaaggtgcatcttagtatctgctttccagaatacactgggccaaatacatttgaagatgca' ggaaactacatcaagaaccagtttctagacctgaatttaaaaaaagaagataaggaaatttattcccacatgacctgtgctactgacacc caaaatgtcaagtttgtgtttgacgcagttacagatataataatcaaagagaatctaaaagactgtgggcttttc (SEQ ED NO: 89)

MOOSE05621 ctgl5361 27306427-27307048, 27307103-27307314,

27309374..27309499

SPGSRVILYΓVFGFGAVLAVFGNLLVMISILHFKQLHSPTNFLVASLACADFLV

GVTVMPFSMVRTVESCWYFGRSFCTFHTCCDVAFCYSSLFHLCFISIDRYIAVTDPLV

YPTKFTVSVSGICISVSWILPLMYSGAWYTGVYDDGLEELSDALNCIGGCQTVVNQ NWVLTDFLSFFIPTFIMΠLYGNTFLVARRQAKKFFINTERKAAKTLGVTVNAFMISWLP YSROSLIDAFMGFITPACRΓΕICCWCAYYNSAMNPLIYALFYPWFPJ_ IKKLRKCKVF REYENKTTRLGWGHACNPSTLGGQGGWITRSKD (SEQ ED NO: 92) tcgccgggatcccgggtgattctgtacatagtgtttggctttggggctgtgctggctgtgtttggaaacctcctggtgatgattt caatcctccatttcaagcagctgcactctccgaccaattttctcgttgcctctctggcctgcgctgatttcttggtgggtgtgactgtgatgc ccttcagcatggtcaggacggtggagagctgctggtattttgggaggagtttttgtactttccacacctgctgtgatgtggcattttgttact cttetctctttcacttgtgcttcatctccatcgacaggtacattgcggttactgaccccctggtctatcctaccaagttcaccgtatctgtgtca ggaatttgcatcagcgtgtcctggatcctgcccctcatgtacagcggtgctgtgttctacacaggtgtctatgacgatgggctggaggaa ttatctgatgccctaaactgtataggaggttgtcagaccgttgtaaatoaaaactgggtgttgacagattttctatccttctttatacc attatgataattctgtatggtaacatatttcttgtggctagacgacaggcgaaaaagatagaaaatactgagagaaaagcagctaaaacc ctgggggtcacagtggtagcatttatgatttcatggltaccalatagcattgattcattaattgatgcctttatgggctttataacccctgcctg tatttatgagalttgctgttggtgtgcttattataacteagccatgaatcctttgatttatgctttatttt^ aaactgagaaagtgcaaagtgtttagggaatatgaaaataaaacaactaggctgggcgtggtgggtcacgcctgtaatcccagcacttt gggaggccaaggcgggtggatcacaaggtcaaaagat (SEQ ED NO: 91) MOOSE05712 ctg3235 322845-323688, 338849..338916

CYNQTLSFTVLTCIISLVGLTGNAVΥLWLLGYRMRRNAVSIYILNLAAADFLF LSFQΠRLPLRLIMSHLIRKILVSV-_TFPYFTGLSMLSAISTERCLS\T.WPIWYRCRRPT HLSAWCVLLWGLSLLFSMLEWRFCDFLFSGADSSWCETSDFIPVAWLIFLCWLCV SSLVLLVRILCGSRKMPLTRLYVΉLLTVLWLLCGLPFGILGALIYRMHLNLEVLYCH VYLVCMSLSSLNSSANPΠYFFVGSFRQRQNRQNLKLVLQRALQDKPEVDKASATRS RTRTTSTSSASTPPRPT (SEQ ED NO: 94) tgctacaatcagaccctgagcttcacggtgctgacgtgcatcatttcccttgtcggactgacaggaaacgcggttgtgctctg gctcctgggctaccgcatgcgcaggaacgctgtctccatctacatcctcaacctggccgcagcagacttcctcttcctcagcttccagat tatacgtttgccattacgcctcatcaatatcagccatctcatccgcaaaatcctcgtttctg^gatgacctttccctactttacaggcctgagt atgctgagcgccatcagcaccgagcgctgcctgtctgttctgtggcccatctggtaccgctgccgccgccccacacacctgtcagcgg tcgtgtgtgtcctgctctggggcctgtccctgctgtttagtatgctggagtggaggttotgtgacttcctgtttagtggtgctgattctagtt^ gtgtgaaacgtcagatttcatoccagtcgcgtggctgatttttttatgtgtggttctctgtgtttccagcctggtcctgctggtcaggatccte^ gtggatcccggaagatgccgctgaccaggctgtacgtgaccatcctgctcacagtgctggtcttcctcctctgcggcctgcccttcggc attctgggggccclaatttacaggatgcacctgaatttggaagtcttatattgtcatgtttatctggtttgcatgtccctgtcctctctaaacag tagtgccaaccccatcatttacttcttcgtgggctcctttaggcagcgtcaaaataggcagaacctgaagctggttctccagagggctct gcaggacaagcctgaggtggataaagcatctgcaacgaggagccggacgagaaccacctcaacctcatccgcgtcaacgccacca aggcctacg (SEQ ED NO: 93)

MOOSE05732 ctg3235 1189276..1189408, 1206579..1207366 CGKETLIP LILFIALVGLVGNGFVLWLLGFP-VIRRNAFSVYVLSLAGADFLF

LCFQIINCLVYLSNFFCSISINFPSFFTTv-_TCAYLAGLSMLSTVSTERCLSVLWPIWYR CRRPRHLSAWCVLLWALSLLLSILEGKFCGFLFSDGDSGWCQTFDFITAAWLIFLFM VLCGSSLALLVMLCGSRGLPLTRLYLTILLTVLWLLCGLPFGIQWFLILWIWKDSDV LFCHIHPVSVVLSSLNSSANPIIYFFVGSFRKQECSLSCLSKQKSPTLLSERDFWLAVIA EVSRGRRHEGFTLSFLG (SEQ ED NO: 96) tgtggcaaggagaccctgatcccggtcttcctgatccttttcattgccctggtcgggctggtaggaaacgggtttgtgctctg gctcctgggcttccgcatgcgcaggaacgccttctctgtctacgtcctcagcctggccggggccgacttcctcttcctctgcttccagatt ataaattgcctggtgtacctcagtaacttcttctgttccatctccatcaatttccctagcttcttcaccactgtgatgacctgtgcctaccttgc aggcctgagcatgctgagcaccgtcagcaccgagcgctgcctgtccgtcctgtggcccatctggtatcgctgccgccgccccagaca cctgtcagcggtcgtgtgtgtcctgctctgggccctgtccctactgctgagcatcttggaagggaagttctgtggcttcttatttagtgatg gtgactctggttggtgtcagacatttgatttcatcactgcagcgtggctgatttttttattcatggttctctgtgggtccagtctggccctgctg gtcaggatcctctgtggctccaggggtctgccactgaccaggctgtacctgaccatcctgctcacagtgctggtgttcctcctctgcggc ctgccctttggcattcagtggttcctaatattatggatctggaaggattctgatgtcttattttgtcatattcatccagtttcagttgtcctgtcat ctcttaacagcagtgccaaccccatcatttacttcttcgtgggctcttttaggaagcaggagtgctcactctcctgtttaagcaagcagaag tctcccaccctcctttcagagagagacttctggattgcagttattgctgaagtaagccgaggaagaagacacgaggggttcacactttcc tttttaggc (SEQ ED NO: 95)

MOOSE05740 ctg3235 274265..275065, 285148..285258 CYKQTLSFTGLTCIVSLVALTGNAVVLWLLGCRMRRNAVSIYILNLVAADFL

FLSGHΠCSPLP INIRIJJISKILSPVMTFPY IGLSMLSAISTERCLSILWPIWYHCRRPR YLSSVMCVLLWALSLLRSILEWMFCDFLFSGANSVWCETSDFITIAWLVFLCWLCG

SSLVLLVRTLCGSRKMPLTRLYVTILLTVLVFLLCGLPFGIQWALFSRIHLDWKVLFC HVHLVSIFLSALNSSANPIIYFFVGSFRQRQNRQNLKLDSMCRRTALYKTIRSRESYSL SREQQREDPTHDSILS (SEQ ED NO: 98) tgctacaagcagaccctgagcttcacggggctgacgtgcatcgtttcccttgtcgcgctgacaggaaacgcggttgtgctct ggctcctgggctgccgcatgcgcaggaacgctgtctccatctacatcctcaacctggtcgcggccgacttcctcttccttagcggccac atta g tcgccg tacgcctcatcaatøtccgccatcccatctccaaaatoctcagtoctgtgatgacctttccctactttataggcctaa gcatgctgagcgccatcagcaccgagcgctgcctgtccatcctgtggcccatctggtaccactgccgccgccccagatacctgtcatc agtcatgtgtgtcctgctctgggccctgtccctgctgcggagtatcctggagtggatgttctgtgacttcctgtttagtggtgctaattctgtt tgg g gaaacgtcagatttcattacaatcgcgtggctggtttttttatgtgtggttctctgtgggtccagcctggtcctgctggtcaggattc tctgtggatcccggaagatgccgctgaccaggctgtacgtgaccatcctcctcacagtgctggtcttcctcctctgtggcctgccctttgg cattcag gggccctgttttocaggatccacctggattggaaagtcttattttgtcatgtgcatctagtttccattttcctgtccgctctta gcagtgccaaccccatcatttacttcttcgtgggctcctttaggcagcgtcaaaataggcagaacctgaagctggacagcatgtgcagg agaacggccctttataaaaccatcagatctcgtgagagttattcactatcacgagaacagcagagggaagacccaacccatgattcaat actttcc (SEQ ED NO: 97)

MOOSE05748 ctg4256 14884996..14885118, 14904164..14905015

HSPTHTFLFFLVLAIFSVAFMGNSVMVLLIYLDTQLHTPMYFLLSQLFLMDLM LICSTVPKMAFNYLSGSKSISMAGCATQEFFYVSLLGSECFLLAVMSYDRYIAICHPLR YTNLMRPKICGLMTAFSWILGSMDAΠDAVATFSFSYCGSREIAHFFCDFPSLLILSCN DTSIFEKVLFICCIVMIWPVAIΠASYARVILAVIHMGSGEGRRKAFTTCSSHLMVVG MYYGAGLFMYIRPTSDRSPMQDKLVSVFYTILTPMLNPLIYSLRNKEVTRALRKVRG APLERKHSDSGFSTSSRSTGCATFSLCAHLRATSSAEV (SEQ ED NO: 100) cacagccccacccacaccttcctcttctttctggtcctggccatcttttcagtggccttcatgggaaactctgtcatggttctcct catctacctggacacccagctccacacccccatgtacttcctcctcagtcaactgttcctcatggacctcatgctcatctgctctaccgtac ccaagatggccttcaactacttgtctggcagcaagtccatttctatggctggttgtgccacacaaattttcttctatgtatcactgcttggctc cgaatgctttctgttggctgttatgtcttatgaccgctatattgccatttgccaccctctaagatacaccaatctcatgagacccaaa atttgtggacttatgactgccttctcctggatcctgggctctatggatgcaatcattgatgctgtagcgacattttccttctcctactgtgggtc tcgggaaatagcccacttcttctgtgacttcccttccctactaatcctctcatgcaatgacacatcaatatttgaaaaggttcttttcatctgct gtatagtaatgattgtttttcctgttgcaatcatoatcgcttcctatgctcgagttattctggctgtcattcacatgggatctggagagggtcgt cgcaaagcttttactacctgttcctctcacctcatggtggtgggaatgtactatggagcaggtttgttcatgtacatacggcccacatctga tcgctcccctatgcaggacaagctggtgtctgtattctacaccatcctcactcccatgctgaatcccctcatctacagcctccgcaacaag gaggtgaccagagcactcaggaaagtgagaggagccccgctggaaaggaaacattcggactccggtttcagcacctcctccagaa gcacaggatgtgcgaccttcagcctttgtgcccacctccgagccaccagctccgcagaagta (SEQ ED NO: 99)

MOOSE05757 ctg30162 475149..475607, 475650-476006, 490932-491042 RWELQIFFFVTFSLIYGATVMGNILEviVTVTCRSTLHSPLYFLLGNLSFLDMCL

STATTPK.MπDLLTDFiKΗSVWGCVTQMFFMHFFGGAEMTLLπMAFDRYVAICKPLH YRT SHKLLKGFAILSWπGFLHSISQIVLTMNLPFCGHNLACIETYTLELFVIADSGL LSFTCFILLLVSYIVILVSVPKKSSHGLSKALSTLSAHirvΥTLFFGPCIFIYVWPFSSLAS NKTLA YTVITPLLNPSIYTLRNKKMQEAII_KLRFQYNSSPSKTFPQTRRJ_DKDYPR KTRAKEFFfϊΗQIFFIRN (SEQ ED NO: 102) cgatgggaacttcaaattttcttctttgtgacattttccctgatctacggtgctactgtgatgggaaacattctcattatggtcaca gtgacatg^ggtcaacccttcattctcccttgtactttctccttggaaatctctcttttttggacatgtgtctctccactgccacaacacccaa gatgatcatagatttgctcactgaccacaagaccatctctgtgtggggctgcgtgacccagatgttcttcatgcacttctttgggggtgctg agatgactcttctgataatcatggcctttgacaggtatgtagccatatgtaaacccctgcactataggacaatcatgagccacaagctgct aaaggggtttgcgatactttcatggataattggttttttacactccataagccagatagttttaacaatgaacttgcctttctgtggccacaat cttgcttgcattgaaacatacaccctggaattatttgtcattgctgacagcgggctgctctctttcacctgtttcatcctcttgcttgtttcttac attgtcatcctggtcagtgtaccaaaaaaatcatcacatgggctctccaaggcgctgtccacattgtctgcccacatcattgtggtcactct gttctttggaccttgtatttttatctatgtttggccattcagtagttt^ ctgaatccgagtatttataccctgagaaataagaaaatgcaagaggccataagaaaattacggttccaatataatagttcacccagcaaa actttccctcagacacgaagaagagataaagactatcccagaaaaacaagagctaaggaatttcattaccaccaaatcttttttataagaa ac (SEQ ED NO: 101)

MOOSE05798 ctgl3495 208823..208941, 209823..210141, 210181..210672

SQDIQLLWVTILIFYLΠLPGNFLΠFTIRSDPGLTAPLYLFLGNLAFLDASYSFIV APRMLVDFLSEKKVISYRGCITQLFFLHFLGGGEGLLLVVMAFDRYIAICRPLHCSTV

MNPRACYAMMLALWLGGFVHSΠQWLILRLPFCGPNQLDNFFCDVRQLLMVFNSG

LMTLLCFLGLLASYAVILCHVRRAASEGKNKAMSTCTTRVΠILLMFGPAIFIYMCPFR

ALPADKMVSLFHTVIFPLMNPMIYTLRNQEVKTSMKRLLIRCLVLCRLMTQTHTRSG

LRNRKFIRQEGREELPHTEGEA (SEQ ED NO: 104) tctcaagatattcagctcttggtctttgtgctgatcttaatrttetaccttatcatcctocctggaaattttotcattattttcac ggtcagaccctgggctcacagcccccctctatttatttctgggcaacttggccttcctggatgcatcctactccttcattgtggctcccagg atgttggtggacttcctctctgagaaaaaggtaatctcctacagaggctgcatcactcagctctttttcttgcacttccttggaggagggga gggattactccttgttgtgatggcctttgaccgctacatcgccatctgccggcctctgcactgttcaactgtcatgaaccctagagcctgct atgcaatgatgttggctctgtggcttgggggttttgtccactccattatccaggtggtcctcatcctccgcttgcctttttgtggcccaaacc agctggacaacttcttctgtgatgtccgacagcttctaatggtcttcaacagtggcctgatgacactcctgtgctttctggggcttctggctt cctatgcagtcatcctctgccatgttcgtagggcagcttctgaagggaagaacaaggccatgtccacgtgcaccactcgtgtcattattat acttcttatgtttggacctgc cttcatctacatgtgccctttcagggccttaccagctgacaagatggtttctctctttcacacagtgatctt tccattgatgaatcctatgatttatacccttcgcaaccaggaagtgaaaacttccatgaagaggttattgattcgttgtcttgtgctgtgccg attaatgacacagactcacacacggagtgggttaaggaacagaaagtttattaggcaagaaggaagagaagagcttccccatacaga gggagaagca (SEQ ED NO: 103)

MOOSE05802 ctg832 2973509..2973633, 2988359-2988723, 2988742..2989221

LEAAHΓ^SIPFCVVYLLALLGNGSLLFΠKTEPSLHEPMYLFLCMLAVVDLVV

CSTAWKLLSLFWFHDGEIRFETCLSLVFLIHSCSTMESGFFLAMAFDRYVAICNPLRH SAILTP^VIGRVGLAIVLRGIALLSPHSFLLRWLPYCRTHIISHTYCEIACAETKFRRAY SLIVAFLTGVVDFILIIYSYVLILHTVFQLPSKDARLKSLGTCGSHVCVILVSYTPAFFS FLTHRFGHHVAJ⁾HFHIFVA YLLWPMVNPIIYGVRTKRIWDI_FLK RLNFQCHff ITDHKRLSπSSFPIKEAACC KQRFCMKLCRR (SEQ ED NO: 106) ttggaagctgctcacatctggatctccatccccttttgtgtggtctacctgttggccctactgggaaacggctctcttctgtttatc atcaagacagagcccagcctccatgagccaatgtacctcttcctatgcatgctggctgtagttgatcttgttgtgtgttctacagctgtgcc caaacttctcagtctcttctggttccatgatggagagattcgctttgaaacctgcctctcactcgtgttcctgattcactcttgctccaccatg gaatctggcttcttcctggccatggcttttgaccgatatgtggccatttgcaatccattaagacattcagctattctgacacgcgctgtaatt gggagagtgggcctagctattgttctcaggggcatagcacttctcagtcctcactctttcctactacgctggcttccctactgcagaaccc atatcatttctcacacctactgtgagattgcctgtgctgagacaaaattccgcagagcctacagcctcattgttgccttccttactggggtg gtagactttatattgatcatttattcttatgtcctcatactccacactgtcttccagctcccatccaaagatgcccggctcaaatctttgggcac ctg^ggctcccatgtctgtgtcatcttag atcctatactccagccttcttctcgtttctcacccacaggtttgggcaccatgtggctccccat tttcacatatttgtggccaacatctatcttcttgtcccacccatggtgaaccccattatctatggggtaagaaccaaaaggatttgggacag gttcctlaaag tttcaggttaaatttccagtgccacatatttgtaatcactgatcataaaagactcagtataatatcttctttcc agcagcctgctgcaaacaaaggttctgcatgaagttatgcaggcga (SEQ ED NO: 105)

MOOSE05811 ctgl5944 2627738..2628080, 2633616-2633643, 2638652..2639093,

2664854..2664991

NPEMNWLSVLFLLIYLITVLGNFWIIIIILASAQLHSPMYFFLSQLAFLDFCYSS VLIPKMLVNYIAGQKVTSYHGCLLQYSFVSLFLTTECFLLAAMACDRYLAVCHPLHY

KEHRVQEPIRCSLISVWVISSLAFCDSSINHFFCDTTALLALSCVDTFGTEMVSFVLAG

FTLLSSLLΠTVTYΠIISAILRIQSAAGRQKAFSTCASHLMAVTIFYGSLIFTYLQPDNTS

SLTQAQVASVFYTIVIPMLNPLIYSLRNKDVKNALLRCSGAVSAHCKLCLLNSRHSPA

SASQVAGTTGAPJfflARLFFLFLVET (SEQ ED NO: 108) aaccctgaaatgaatgttgtcctttctgtgctctttetattaatctatotcattactgtcttgggcaacttttggattatcataataattc tggctagtgcccaactccattcacccatgfactttttccttagccagftggcltt^ ggtgaattacatagcaggacagaaagtcatctcttatcacggttgcctccttcagtattcctttgtcagcttgttcctgactactgaatgcttc ctcctggctgccatggcatgtgatcggtatctcgctgtttgccacccacttcactacaaagaacatagagtccaggagccaattagatgc tcgctgatatctgtctgggtgataagcag ttggcgttctgtgattccagcatcaatcattttttttgtgacaccacagctcttttagcactctc ctgtgtagatacattcggcacagaaatgg gagctttgtcttagctggattcactcttcttagctctctccttatcatcacagtcacttatatca tcatcatctcagccatcctgaggatccagtcagcagcaggcaggcagaaggccttctccacctgcgcatcccacctcatggctgtaact atcttttatgggtctctgattttcacctatttgcaacctgataacacatcategctgacccaggcgcaggtggcatctgtattctatacgattg tcattcccatgctgaatccactcatctacagtctgaggaacaaagatgtgaaaaatgctcttctgagatgcagtggtgccgtctcggctca ctgcaagctctgcctcctgaattcacgccattctcctgcctcagcctcccaagtcgctgggactacaggcgcccgccaccacgcaagg ctattttttttatttttagtagagacg (SEQ ED NO: 107)

MOOSE05812 . ctg4256 14956033..14956176, 14958553-14958878, 14958906-14959383

QSmGLFVFTLIFLIFLMALIGNLSMILLIFLDIHLHTPMYFLLSQLSLIDLNYISTI WKMVYDFLYGNKSISFTGCGIQSFFFLTLAVAEGLLLTSMAYDRYVAICFPLIIYPIRI SKRVCVMMITGSWMSSINSCAHTVYALCIPYCKSP^ ivIHFFSCTDTWVYESTVFLSS TIFLVLPFTGIACSYGRVLLAVYRMHSAKGRNKVYSTCSTHLTVVYFYYAPFAYTYV P ^RSLRSPTEDKILAWYΗLTPMLNPπYSLPNKESFCTAKETTmVNRQPTEWEKIFA TYSSDKGLISPJYNELKQTYKKKT (SEQ ED NO: 110) caatcaagaattggccttttcgtattcacccteatttttctcattttcctaatggctctaattggaaatctatccatgattcttctcatct ttttggacatccatctccacacacctatgtatttcctacttagtcagctctccctcattgacctaaattacatctccaccattgttccaaagatg gtttatgaltttctg^tggaaacaag ctatctccttcactggatgtgggattcagagtttcttcttcttgactttagcagttgcagaagggct gctcctgacatcaatggcctatgatcgttatgtggccatttgctttcctctccactatcccatccgtataagcaaaagagtgtgtgtgatgat gataacaggatcttggatgataagctctatcaactcttgtgctcacacagtatatgcactctgtatcccatattgcaagtccagagccatca atcattttttctcctgcacagacacttgggtctatgagagcacagtgtttttgagcagcaccatctttcttgtgcttcctttcactggtattgcat gttcctatggccgggttctccttgctgtctaccgcatgcactctgcaaaagggaggaataaggtctattcaacctgtagcacccacctca ctg ggtglacttctactatgcaccctttgcttatacctatgtacgtgcaagatccctgcgatctccaaccgaggacaagattctggctgttt tctacaccatcctcaccccaatgctcaaccccatcatctacagcctgagaaacaaggagagcttctgcacagcaaaagaaactaccat cagagtgaacaggcaacctacagaatgggagaaaatttttgcaacctactcatctgacaaagggctaatatccagaatctacaatgaac tcaaacaaacttacaagaaaaaaaca (SEQ ED NO: 109)

MOOSE05827 ctg4256 15055783-15055926, 15062025-15062324, 15062361..15062855

PSPJDLFFFILIVFIFLMALIGNLSMILLIFLDTHLHTPMYFLLSQLSLIDLNYISTI VPKMASDFLHGNKSISFTGCGIQSFFFLALGGAEALLLASMAYDRYIAICFPLHYLIR MSKRVCVLMITGSWIIGSINACAHTVYVLHIPYCRSRAINHFFCDVPAMGTVFLSATI FL PFIGISCSYGQVXFAVΥHMKSAEGRKKAYLTCSTHLTVVTFYYAPFVYTYLRP RSLRSPTEDKVLAVFYTILTPMLNPΠYSLR^KESKRKKRKERJ_RKRKKRKKERERK^

EREKEKERKERKEREKERKERKKE (SEQ ED NO: 112) ccatcaagaattgaccttttcttcttcattctcattgttttcattttcctgatggctctaattggaaacctgtccatgattcttctcatctt cttggacacccatctccacacacccatgtatttcctactgagtcagctctccctcattgacctaaattacatctccaccattgttcctaagat ggcatctgattttctgcatggaaacaagtctatctccttcactgggtgtgggattcagagtttcttcttcttggcattaggaggtgcagaagc actacttttggcatctatggcctatgatcgttacattgctatttgctttcctctccactatctcatccgcatgagcaaaagagtgtgtgtgctga tgataacagggtcttggatcataggctcgatcaatgcttgtgctcacactgtatatgtactccatattccttattgccgatccagggccatca atcatttcttctgtgatgtcccagcaatgggcacagtgtttttgagtgccaccatctttctcgtgtttcccttcattggtatttcatgttcctatgg ccaggttctctttgctgtctaccacatgaaatctgcagaagggaggaagaaagcctatttgacctgcagcacccacctcactgtagtaac tttctactatgcaccttttgtctacacttatctacgtccaagatccctgcgatctccaacagaggacaaggttctggctgtcttctacaccatc ctcaccccaatgctcaaccccatcatctatagcctgaggaacaaggagagcaagagaaagaaaagaaaggaaagaaagagaaaaa gaaagaaaagaaagaaagaaagagaaagaaagagagaaagagagaaagagaaagaaagaaaagaaaggaaagaaagagaaa aagaaagaaaagaaagaaagaaagag (SEQ ID NO: 111)

MOOSE05861 ctgl5944 6729088..6729914, 6731331..6731469 SRELSQVLFTFLFLVYMTTLMGNFLIMVΓV CESHLHTPMYFLLRNLSILDICF

SSITAPKVLIDLLSETKΉSFSGCVTQMFFFHLLGGADVFSLSVMAFDRYIAISKPLHY MTIMSRGRCTGLIVGFLGGGLVΉSIAQISLLLPLPVCGPNVLDTFYCDVPQVLKLACT DTFTLELLMISNNGLVSWFVFFFLLISYTVILMMLRSHTGEGRRKAISTCTSHITWTL HFWCIΎVYARPFTALPTDTAISVTFTVISPLLNPΠYTLRNQEMNNRSRHLSKKKKKR KDKKKPACAJ__ENEVRLCFFLCVTVNSVQPWRNT (SEQ ED NO: 114) tcccgagaactgagccaggtcttatttaccttcctgtttttggtgtacatgacaactctaatgggaaacttcctcatcatggttac agttøcctgtgaatctcaccttcatacgcccatgtacttcctgctccgcaacctgtctattcttgacatctgcttttcctccatcacagctccta aggtcctgatagatcttctatcagagacaaaaaccatctccttcagtggctgtgtcactcaaatgttcttcttccaccttctggggggagca gacgttttttctctctctgtgatggcgtttgaccgctatatagccatctccaagcccctgcactatatgaccatcatgagtagggggcgatg cacaggcctcatcgtgggcttcctgggtggggggcttgtccactccatagcgcagatttctctattgctcccactccctgtctgtggaccc aatgttcttgacactttctactgcgatgtcccccaggtcctcaaacttgcctgcactgacaccttcactctggagctcctgatgatttcaaat aatgggttagtcagttggtttgtattcttctttctcctcatatcttacacggtcatcttgatgatgctgaggtctcacactggggaaggcagga ggaaagccatctccacctgcacctcccacatcaccgtggtgaccctgcatttcgtgccctgcatctatgtctatgcccggcccttcactg ccctccccacagacactgccatctctgtcaccttcactgtcatctcccctttgctcaatcctataatttacacgctgaggaatcaggaaatg aacaacagatcaagacacttgtcaaaaaaaaaaaagaaaagaaaagacaaaaagaaaccagcatgcgcaaagaaagaaaatgagg tgagattgtgccacatttgtgtaacagtgaactcagtgcagccatggcgtaataca (SEQ ED NO: 113)

MOOSE05863 ctgl5907 31055206-31056122, 31068740-31068797

WQQQQVLLFALFLCLYLTGLFGNLLILLAIGSDHCLHTPMYFFLANLSLVDLC LPSAT KMLLNIQTQTQTISYPGCLAQMYFCMMFANMDNFLLTVMAYDRYVAICH PLHYSTIMALRLCASLVAAPWVIAILNPLLHTLMMAHLHFCSDNVTHHFFCDINSLLP LSCSDTSLNQLSVLATVGLIFVWSVCILVSYILIVSAVMKVPSAQGKLKAFSTCGSHL ALVILFYGAITGλ viSPLSNHSTEKDSAASVIFMVVAPVLNPFIYSLRNNELKGTLKK TLSRPGAVAHACNPSTLGGRGGNTWTEAKYVT__EvT_ IKR (SEQ ED NO: 116) tggcaacaacagcaggtgctactctttgcacttttoctgtgtctctatttaacagggctgtttggaaacttactcatcttgctggc cattggctcggatcactgccttcacacacccatgtatttcttccttgccaatctgtccttggtagacctctgccttccctcagccacagtccc caagatgctactgaacatccaaacccaaacccaaaccatctcctatcccggctgcctggctcagatgtatttctgtatgatgtttgccaat atggacaattttcttctcacagtgatggcatatgaccgttacgtggccatctgtcaccctttacattactccaccattatggccctgcgcctc tgtgcctctctggtagctgcaccttgggtcattgccattttgaaccctctcttgcacactcttatgatggcccatctgcacttctgctctgata atgttatccaccatttcttctgtgatatcaactctctcctccctctgtcctgttccgacaccagtcttaatcagttgagtgttctg gctacggtggggctgatctttg^ggtaccttcagtg gtatcctggtatcctatatcctcattgtttctgctgtgatgaaagtcccttctgccc aaggaaaactcaaggctttctctacctgtggatctcaccttgccttggtcattcttttctatggagcaatcacaggggtctatatgagcccct tatccaatcactctactgaaaaagactcagccgcatcagtcatttttatggttg agcacctgtgttgaatccattcatttacagtttaagaaa caatgaactgaaggggactttaaaaaagaccctaagccgaccgggcgcggtggctcacgcctgtaatcccagcactttgggaggcc gaggcggaaatacatggacagaagcaaaatatgttcatagagaagtacatataatgatcaagaga (SEQ ID NO: 115)

MOOSE05908 ctgl5944 2316216..2316693, 2331854-2331911, 2333360-2333372,

2338898-2339200, 2350975-2351085 YPEIQWLFLWLFVYTVWVGNLGMNILRLNSKLHTIMCFFLSHLSLTDFCFST

WTPKLLENLWEYRTISFSGCIMQFCFACIFGVTETFMLAAMAYDRFVAVCKPLLY TTMSQKLCALLVAGSYTWGIVCSLILTYFLLDLSFCESTFINNFISFNSTLYYKΓYHQT FSCICEOTLPMCTLLΠLTSYV IJVTVLKIRSVSGRHKAFSTWASHLTSITIFHGTILFLY C NSKNSRQTVKVASWYTVVN MLNPLIYSLRNKDVTODAFWKLIHTQSRRRQSL RRSLRNSHLMTTSIEEGVTHTSTILELCSS (SEQ ED NO: 118) tacccagaaatccaggttccactctttctggttttcttgttcgtctacacagtcactgtagtggggaacttgggcatgataataat catcagactcaattcaaaactccatacaatcatgtgctttttocttagtcacttgtccttgacagacttctgtttttccactgtagttacacctaa actgttggagaacttggttgtggaatacagaaccatctctttctctggttgcatcatgcaattttgttttgcttgcatttttggagtgacagaaa ctttcatgttagcagcgatggcttatgaccgttttg ggcagtttgtaaacccttgctgtataccactattatgtctcagaagctotgtg^ ctggtggctgggtoctatacatgggggatagtgtgctccctgatactcacatattttcttcttgacttatcgttttgtgaatctaccttcataaat aattttatctctttcaactccaccttatactataaaatataccaccaaaccttctcctg^atatgtgagcactttctgccaatgtgtacactactg atcatcctcacttcctatgttttcatttttgtgactgtactaaaaatccgttctgttagtgggcgccacaaagccttctccacctgggcctccc acctgacttctatcaccatcttccatgggaccatccttttcctttactgtgtacccaactccaaaaactctcggcaaacagtcaaagtggcc tctgtattttacacagttgtcaaccccatgctgaaccctctgatctacagcctaaggaataaagacgtgaaggatgctttctggaagttaat acatacacaatcaagaaggaggcagagcttgaggaggtccttgagaaattcccatttaatgaccaccagcattgaagaaggagtcacc cacaccagtaccattctggagctctgctcatcc (SEQ ED NO: 117)

MOOSE05917 ctg30162 1982852..1982944, 1985177..1985529, 1985575..1986055

PLRLRTLFFWFFLIYILTQLGNLLILITV VADPRLHARPMYIFLGVLSVIDMSIS SIIVPP MMNFTLGV- PIPFGGCVAQLYFYHFLGSTQCFLYTLMAYDRYLAICQPLRY

PVLMTAKLSALLVAGAWMAGSIHGALQAILTFRLPYCGPNQVDYFFFNELVTFVDIG

VVVASCFSLILLSYIQΠQ LRIHTADGRRRAFSTCGAHVTVVTVYYVPCAFIYLRPET

NSPLDGAAALVPTAITPFLNPLIYTLRNQEVKLALKRMLRSPRTPSEHFGRPRRVDHL

RSGVRDQPNQHGETASLPKI (SEQ ED NO: 120) ccactcaggctaaggacactcttttttgtgflcttttttctaatctacatcctgactcagctgggaaacctgcttattttaatcactgt ctgggcagacccaaggctccatgcccgccccatgtacatctttcttggtgttctctcagtcattgatatgagcatctcctccatcattgtccc tcgcctcatgatgaacttcactttaggtgtcaaacccatcccatttggtggctgtgttgctcaactctatttctatcacttcctgggcagcacc cagtgcttcctctacaccctaatggcctatgacaggtacctggcaatatgtcagcccctgcgctaccctgtgctcatgactgctaagctg agcgccttgcttgtggctggagcctggatggcaggatccatccatggggctctccaggccatcctaaccttccgcctgccctactgtgg gcccaatcaggtggattacttcttcttcaacgagctggtgacgtttgtagacattggggtggtggttgccagttgcttctccctgatcctcct ctcctacatacagatcattcaggccatcctgagaatccacacagctgatgggcggcgccgggctttttcaacttgtggagcccatgtaa ccgtggtcaccgtgtactatgtgccctgtgccttcatctacctgaggcctgaaaccaacagccccctggatggggcagctgccctagtc cccacggccatcactcctttcctcaacccccttatctacactctgcggaaccaagaggtgaagctggccctgaaaagaatgctcagaa gcccaagaactccgagtgagcactttgggaggccaaggcgggtggatcacctgaggtcgggagttcgagaccagcctaaccaacat ggagaaactgcatctttaccaaaaata (SEQ ED NO: 119)

MOOSE06051 ctgl5064 38069881-38070569, 38071866-38071965, 38089519-38089577, 38100385..38100508

DPTV^'TPHLISLYFIVLIGGLVGVISILFLLVKMNTRSVTTMAVINLVVVHSVFLL

TVPFRLTYLITXTWMFGLPFCKFVSAMLMHMYLTFLFYVVILVTRYLIFFKCKDKVE FYRJ_LHAVAASAGMWTLVRVTVNPLVΥSRYGIHEEYNEEHCFKFHKELAYTYVKIΙN YMIVIFVIAVAVILLVFQVFΠMLMVQKLRHSLLSHQEFWAQLKNLFFIGVILVCFLPY QPHCVMFPSLCPCVFIVQLPLLSENMRCLVFCSCLIHFLLYΠ^?IVHNMRYSYMYNFVS CPLPACNAVMQCSGSNLEQSRKQYSQCSRPGTARGKL (SEQ ED NO: 122) gatcctatagtgacaccccacttaatcagcctctacttoatag gcttattggcgggctggtggg^gtcatttccattcttttcctc ctggtgaaaatgaacacccggtcagtgaccaccatggcggtcattaacttggtggtggtccacagcgtttttctgctgacagtgccatttc gcttgacctacctcatcaagaagacttggatgtttgggctgcccttctgcaaatttgtgagtgccatgctgcacatccacatgtacctcac gttcctattctatgtggtgatcctggtcaccagatacctcatcttcttcaagtgcaaagacaaagtggaattctacagaaaactgcatgctg tggctgccagtgctggcatgtggacgctggtgattgtcattgtggtacccctggttgtctcccggtatggaatccatgaggaatacaatg aggagcactg tttaaatttcacaaagagcttgcttacacatatgtgaaaatcatcaactatatgatagtcatttttgtcatagccgttgctgt gattctgttggtcttccaggtcttcatcattatgttgatggtgcagaagctacgccactctttactatcccaccaggagttctgggctcagct gaaaaacctattttttataggggtcatccttgtttgtttccttccctaccagccccattgtgtgatgttcccctccctg gtccatgtg tttcatt gttcaactcccacttctaagjgagaacatgcggtgtttggttttctgttcctgtttg gtattcatacatgtataacttcgtcagctgtcctttgccagcatgtaatgcagtgatgcaatgctctggtagtaatttggaacaatccaggaa gcaatattctcagtgttcaaggcccggcactgcccgggggaaatta (SEQ ED NO: 121)

MOOSE07369 ctgl5361 27381078-27381945, 27422885-27422949 HVLNFQELFFLVFGVSTLIVWLMVLΠLTTLVGNLIVIVSISHFKQLHTPTNWLI

HSMATVDFLLGCLVMPYSMVRSAEHCWYFGEVFCKIHTSTDIMLSSASIFHLSFISID

RYYAVCDPLRYKAJ_V LVTCVMIFISWSWAWAFGMIFLELNFKGAEEIYYKHV^ CRGGCS FSKISGVLTFMTSFYIPGSIMLCVYYP YLIAKEQARLISDANQKLQIGLE MI_NGISQSKERKAVKTLGIVMG LICWCPFFICTvTvmPFLHYπPPTLM)vXIΛVFGY LNSTFNPMVYAFFYPWFRKALKM (SEQ ED NO: 124) catgttcttaalttccaagaactcttttttcttgtcttcggagtttcaacacttattg^ actcgttggcaatctgatagttattgtttctatatcacacttcaaacaacttcataccccaacaaattggctcattcattccatggccactgtg gactttcttctggggtgtctggtcatgccttacagtatggtgagatctgctgagcactgttggtattttggagaagtcttctgtaaaattcaca caagcaccgacattatgctgagctcagcctccattttccatttgtctttcatctccattgaccgctactatgctgtgtgtgatccactgagata taaagccaagatgaatatcttggttatttgtgtgatgatcttcattagttggagtgtccctgctglttttgcatttggaatgatctttctggagct aaacttcaaaggcgctgaagagatatattacaaacatgttcactgcagaggaggttgctctgtcttctttagcaaaatatctggggtactg acctttatgacttctttttatatacctggatctattatg tatgtgtctattøcagaatalatcttatogctaaagaacaggcaa atgccaatcagaagctccaaattggattggaaatgaaaaatggaatttcacaaagcaaagaaaggaaagctgtgaagacattggggat tgtgatgggagttttcctaatatgctggtgccctttctttatctg acagtcatggacccttttcttcactacattattccacctøcttt gtattgatttggtttggc„cttgaactctacatttaatccaatggtttatgcatttttctatccttggtttagaaaagcactgaagatg (SEQ ED NO: 123) Table UI ctgl4797_MOOSE00162.xml Angiotensin ctgl6465_MOOSE00638.xml Class A Orphan ctgl6008_MOOSE00693.xml Class A Orphan ctgl 6282_MOOSE00717.xml Class A Orphan ctgl 6228_MOOSE00721.xml Class A Orphan ctg877_MOOSE00741.xml Class A Orphan ctgl 5378_MOOSE00766.xml Class A Orphan ctg78_MOOSE00772.xml Class A Orphan ctgl 5540_MOOSE00775. xml Class A Orphan ctgl6537_MOOSE00779.xml Class A Orphan ctg30162_MOOSE00804.xml Class A Orphan ctgl5378_MOOSE00814.xml Class A Orphan ctgl5907_MOOSE00818.xml Class A Orphan ctgl4294_MOOSE00822.xml Class A Orphan ctgl 5968_MOOSE00826.xml Class A Orphan ctgl4145_MOOSE00829.xml Class A Orphan ctgl4667_MOOSE00838.xml Class A Orphan ctgl4333_MOOSE00841.xml Class A Orphan ctgl 5064_MOOSE00843.xml Class A Orphan ctgl 5944_MOOSE00846.xml Class A Orphan ctgl4333_MOOSE00855.xml Class A Orphan ctgl 6279_MOOSE00861.xml Class A Orphan ctgl 8147_MOOSE00872.xml Class A Orphan ctgl5944_MOOSE00880.xml Class A Orphan ctgl5944_MOOSE00882.xml Class A Orphan ctgl5296_MOOSE00886.xml Class A Orphan ctgl7659_MOOSE00899.xml Class A Orphan ctgl 8867_MOOSE00930.xml Class A Orphan ctg3235_MOOSE00941.xml Class A Orphan ctgl 5907_MOOSE00981.xml Class A Orphan ctgl 7802_MOOSE00994.xml Class A Orphan ctgl2634_MOOSE01139.xml Class B Secretin Receptor ctgl5998_MOOSE01146.xml Class B Secretin Receptor ctgl2559_MOOSE01148.xml Class B Secretin Receptor ctgl5361_MOOSE01165.xml Class C Metabotropic Glutamate Receptor ctgl4779_MOOSE01172.xml Class C Metabotropic Glutamate Receptor ctgl7658_MOOSE01176.xml Class C Metabotropic Glutamate Receptor ctgl3655_MOOSE01371.xml Frizzled related GPCR ctgl 734 l_MOOSE01451. xml Gonadotropin receptor ctg37_MOOSE01609.xml Lysosphingolipid Receptor ctgl 5037_MOOSE02359.xml Mollusc Rhodopsine-like GPCR ctgl5907_MOOSE02360.xml Mollusc Rhodopsine-like GPCR

ctgl2376_MOOSE02352.xml unclassified GPCRs ctgl3067_MOOSE01169.xml Class C Metabotropic glutamate pheromone ctgl3284_MOOSE00810.xml Class A Orphan other ctgl4473_MOOSE01192.xml Class Y G proteins ctgl 5361_MOOSE05621.xml Class A O han GPCRs ctg3235_MOOSE05712.xml Class A Orphan GPCRs ctg3235_MOOSE05732.xml Class A Orphan GPCRs ctg3235_MOOSE05740.xml Class A Orphan GPCRs ctg4256_MOOSE05748.xml Class A Orphan GPCRs ctg30162_MOOSE05757.xml Class A Oφhan GPCRs ctgl 3495_MOOSE05798.xml Class A Orphan GPCRs ctg832_MOOSE05802.xml Class A Oφhan GPCRs ctgl 5944_MOOSE05811.xml Class A Oφhan GPCRs ctg4256_MOOSE05812.xml Class A Oφhan GPCRs ctg4256_MOOSE05827.xml Class A Oφhan GPCRs ctgl 5944_MOOSE05861.xml Class A Oφhan GPCRs ctgl 5907_MOOSE05863. xml Class A Oφhan GPCRs ctgl 5944_MOOSE05908.xml Class A Oφhan GPCRs ctg30162_MOOSE05917.xml Class A Oφhan GPCRs ctgl 5064_MOOSE06051.xml Class A Oφhan GPCRs ctgl5361_MOOSE07369.xml Serotonin Gated Ion Channel Receptor

Table IV

OTHER GPCRs

////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////////

RA (Rheumatoid Arthritis)

Locus2 Marker:D6S265 Lod:3 CM RANGE of one LOD drop: 6

MOOSE05863 Class A Oφhan other DISTANCE: -1.97 Mb MOOSE06011 Class A Oφhan other DISTANCE: 8.000 Mb

COPD (Chronic Obstructive Pulmonary Disease)

Locus4 Marker:D19S884 Lod:2.9 CM RANGE of one LOD drop: 20

MOOSE05784 Class A Oφhan other DISTANCE: 0.959 Mb

////////////////////////////////////W

Asthma

Locusl Marker:D3S3698 Lod:3.8 CM RANGE of one LOD drop: 15

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Longevity

Locus 1 Marker:D9S1826 Lod:2.30 CM RANGE of one LOD drop: 15

MOOSE06089 Class B Secretin like DISTANCE: -12.5 Mb

Locus2 Marker:Dl 1 S987 Lod:2.07 CM RANGE of one LOD drop: 12

MOOSE05934 Class A Oφhan other DISTANCE: -14.1 Mb MOOSE05861 Class A Oφhan other DISTANCE: -12.4 Mb MOOSE05761 Class A Oφhan other DISTANCE: -0.76 Mb MOOSE05781 Class A Oφhan other DISTANCE: 0.884 Mb MOOSE05938 Class A Oφhan other DISTANCE: 1.364 Mb

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Psoriasis

Locus 1 Marker:GDB: 182383 Lod:10.61 CM RANGE of one LOD drop: 2 MOOSE05863 Class A Oφhan other DISTANCE: -3.49 Mb

MOOSE06011 Class A Oφhan other DISTANCE: 6.481 Mb

Locus2 Marker:D7S669 Lod: 1.81 CM RANGE of one LOD drop:20 MOOSE06127 Class Y G proteins DISTANCE: 2.208 Mb

Claims

CLAIMS What is claimed is:

1. An isolated nucleic acid molecule comprising a G protein-coupled receptor (GPCR) gene, wherein the GPCR gene has a nucleotide sequence selected from the group of nucleic acid sequences as shown in Tables I and II, or the complements of the group of nucleic acid sequences as shown in Tables I and π.

2. A nucleic acid encoding a polypeptide, wherein the polypeptide has an amino acid sequence selected from the group consisting of the group of amino acid sequences as shown in Tables I and H

3. An isolated nucleic acid molecule which hybridizes under high stringency conditions to a nucleotide sequence selected from the group of nucleic acid sequences as shown in Tables I and π, or the complements of the group of nucleic acid sequences as shown in Tables I and π.

4. An isolated nucleic molecule that hybridizes under high stringency conditions to a nucleotide sequence encoding an amino acid sequence selected from the group consisting of the group of amino acid sequences as shown in Tables I and π.

5. A method for assaying for the presence of a first nucleic acid molecule in a sample, comprising contacting said sample with a second nucleic acid molecule, where the second nucleic acid molecule comprises a nucleotide sequence selected from the group of nucleic acid sequences as shown in Tables I and π, and hybridizes to the first nucleic acid under high stringency conditions.

6. A vector comprising an isolated nucleic acid molecule selected from the group consisting of: (a) the nucleic acid sequences as shown in Tables I and II; (b) the complement of one of the nucleic acid sequences are shown in

Tables I and II; or (c) a nucleic acid encoding an amino acid molecule as shown in Tables I and II; wherein the nucleic acid molecule is operably linked to a regulatory sequence.

7. A recombinant host cell comprising the vector of Claim 6.

8. A method for producing a polypeptide encoded by an isolated nucleic acid molecule, comprising culturing the recombinant host cell of Claim 7 under conditions suitable for expression of the nucleic acid molecule.

9. An isolated polypeptide encoded by the nucleotide sequence of the group of nucleic acid sequences as shown in Tables I and U, or the complements thereof.

10. The isolated polypeptide of Claim 9, wherein the polypeptide has an amino acid sequence selected from the group consisting of the group of amino acid sequences as shown in Tables I and II.

11. An isolated polypeptide comprising an amino acid sequence, wherein the amino acid sequence is greater than about 95% identical to an amino acid sequence selected from the group consisting of the group of amino acid sequences as shown in Tables I and II.

12. A fusion protein comprising an isolated polypeptide of Claim 2.

13. A fusion protein comprising an isolated polypeptide of Claim 11.

14. An antibody, or an antigen-binding fragment thereof, which selectively binds to a polypeptide of Claim 2, or to a fragment or variant of said amino acid sequence.

15. An antibody, or an antigen-binding fragment thereof, which selectively binds to a polypeptide of Claim 11, or to a fragment or variant of said amino acid sequence.

16. A method of assaying for the presence of a polypeptide encoded by an isolated nucleic acid molecule according to Claim 1 in a sample, the method comprising contacting the sample with an antibody which specifically binds to the encoded polypeptide.

17. A method of identifying an agent which alters the activity of a GPCR, the method comprising:

(a) contacting a polypeptide of Claim 9, or a derivative or fragment thereof, with an agent to be tested;

(b) assessing the level of activity of the polypeptide or derivative or fragment thereof; and

(c) comparing the level of activity with a level of activity of the polypeptide or active derivative or fragment thereof in the absence of the agent; wherein if the level of activity of the polypeptide or derivative or fragment thereof in the presence of the agent differs, by an amount that is statistically significant, from the level in the absence of the agent, then the agent is an agent that alters activity of a GPCR.

18. An agent that alters the activity of a GPCR, identifiable according to the method of Claim 17.

19. The agent of Claim 18, where the agent is selected from the group consisting of: a GPCR gene binding agent; a G-protein; a peptidomimetic; a fusion protein; a prodrug; an antibody; and a ribozyme.

20. A method of altering activity of a polypeptide encoded by a GPCR gene, comprising contacting the polypeptide with an agent of Claim 19.

21. A method of identifying an agent which alters interaction of the polypeptide of

Claim 9 with a GPCR gene binding agent, comprising: a) contacting the polypeptide or a derivative or fragment thereof, and the binding agent, with an agent to be tested; b) assessing the interaction of the polypeptide or derivative or fragment thereof with the binding agent; and c) comparing the level of interaction with a level of interaction of the polypeptide or derivative or fragment thereof with the binding agent in the absence of the agent, wherein if the level of interaction of the polypeptide or derivative or fragment thereof in the presence of the agent differs by an amount that is statistically significant, from the level of interaction in the absence of the agent, then the agent is an agent that alters interaction of the polypeptide with the binding agent.

22. An agent that alters interaction of a GPCR polypeptide with a GPCR binding agent, identifiable according to the method of Claim 21.

23. An agent that alters interaction of a GPCR polypeptide with a GPCR binding agent, selected from the group consisting of: a second GPCR binding agent; a G-protein; a peptidomimetic; a fusion protein; a prodrug; an antibody; and a ribozyme.

24. A method of altering interaction of a GPCR polypeptide with a GPCR binding agent, comprising contacting the GPCR gene polypeptide and/or the GPCR gene binding agent with an agent of Claim 23.

25. A method of identifying an agent that alters expression of a GPCR gene, comprising the steps of: a) contacting a solution containing a nucleic acid comprising the promoter region of the GPCR gene operably linked to a reporter gene with an agent to be tested; b) assessing the level of expression of the reporter gene; and c) comparing the level of expression with a level of expression of the reporter gene in the absence of the agent, wherein if the level of expression of the reporter gene in the presence of the agent differs, by an amount that is statistically significant, from the level of expression in the absence of the agent, then the agent is an agent that alters expression of the GPCR gene.

26. An agent that alters expression of the GPCR gene, identifiable according to the method of Claim 25.

27. A method of identifying an agent that alters expression of a GPCR gene, comprising the steps of: a) contacting a solution containing a nucleic acid of Claim 1 or a derivative or fragment thereof with an agent to be tested; b) assessing expression of the nucleic acid, derivative or fragment; and c) comparing expression with expression of the nucleic acid, derivative or fragment in the absence of the agent, wherein if expression of the nucleotide, derivative or fragment in the presence of the agent differs, by an amount that is statistically significant, from the expression in the absence of the agent, then the agent is an agent that alters expression of the GPCR gene.

28. The method of Claim 27, wherein the expression of the nucleotide, derivative or fragment in the presence of the agent comprises expression of one or more splicing variant(s) that differ in kind or in quantity from the expression of one or more splicing variant(s) the absence of the agent.

29. An agent that alters expression of a GPCR gene, identifiable according to the method of Claim 27.

30. An agent that alters expression of a GPCR gene, selected from the group consisting of: antisense nucleic acid to a GPCR gene; a GPCR gene polypeptide; a GPCR gene receptor; a GPCR gene binding agent; a peptidomimetic; a fusion protein; a prodrug thereof; an antibody; and a ribozyme.

31. A method of altering expression of a GPCR gene, comprising contacting a cell containing a GPCR gene with an agent of Claim 30.

32. A method of identifying a polypeptide which interacts with a GPCR gene polypeptide, comprising employing a yeast two-hybrid system using a first vector which comprises a nucleic acid encoding a DNA binding domain and a GPCR gene polypeptide, splicing variant, or a fragment or derivative thereof, and a second vector which comprises a nucleic acid encoding a transcription activation domain and a nucleic acid encoding a test polypeptide, wherein if transcriptional activation occurs in the yeast two- hybrid system, the test polypeptide is a polypeptide which interacts with a

GPCR polypeptide.

33. A GPCR gene therapeutic agent selected from the group consisting of: a

GPCR gene or fragment or derivative thereof; a polypeptide encoded by a GPCR gene; a G-protein; a GPCR gene binding agent; a peptidomimetic; a fusion protein; a prodrug; an antibody; an agent that alters GPCR gene expression; an agent that alters activity of a polypeptide encoded by a GPCR gene; an agent that alters posttranscriptional processing of a polypeptide encoded by a GPCR gene; an agent that alters interaction of a GPCR gene with a GPCR gene binding agent; an agent that alters transcription of splicing variants encoded by a GPCR gene; and a ribozyme.

34. A pharmaceutical composition comprising a GPCR gene therapeutic agent of

Claim 33.

35. The pharmaceutical composition of Claim 34, wherein the GPCR gene therapeutic agent is an isolated nucleic acid molecule comprising a GPCR gene or fragment or derivative thereof.

36. The pharmaceutical composition of Claim 34, wherein the GPCR gene therapeutic agent is a polypeptide encoded by the GPCR gene.

37. A method of treating a disease or condition associated with a GPCR in an individual, comprising administering a GPCR gene therapeutic agent to the individual, in a therapeutically effective amount.

38. The method of Claim 37, wherein the GPCR gene therapeutic agent is a GPCR gene agonist.

39. The method of Claim 38 wherein the GPCR gene therapeutic agent is a GPCR gene antagonist.

40. A transgenic animal comprising a nucleic acid selected from the group consisting of: an exogenous GPCR gene and a nucleic acid encoding a G GPPCCRR g eeennee r p>oollvypr>eeDpttiiddee.

41. A method for assaying a sample for the presence of a GPCR gene nucleic acid, comprising: a) contacting said sample with a nucleic acid comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the sequence of said GPCR gene nucleic acid under conditions appropriate for hybridization, and b) assessing whether hybridization has occurred between a GPCR gene nucleic acid and said nucleic acid comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the sequence of said GPCR gene nucleic acid; wherein if hybridization has occuπed, a GPCR gene is present in the nucleic acid.

42. The method of Claim 41, wherein said nucleic acid comprising a contiguous nucleotide sequence is completely complementary to a part of the sequence of said GPCR gene nucleic acid.

43. The method of Claim 41 , comprising amplification of at least part of said

GPCR gene nucleic acid.

44. The method of Claim 41, wherein said contiguous nucleotide sequence is 100 or fewer nucleotides in length and is either: a) at least 80% identical to a contiguous sequence of nucleotides in one of the nucleic acid sequences as shown in Tables I and II; b) at least 80% identical to the complement of a contiguous sequence of nucleotides in one of the nucleic acid sequences as shown in Tables I and II; or c) capable of selectively hybridizing to said GPCR gene nucleic acid.

45. A reagent for assaying a sample for the presence of a GPCR gene nucleic acid, said reagent comprising a nucleic acid comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the nucleotide sequence of said GPCR gene nucleic acid.

46. The reagent of Claim 45, wherein the nucleic acid comprises a contiguous nucleotide sequence that is completely complementary to a part of the nucleotide sequence of said GPCR gene nucleic acid.

47. A reagent kit for assaying a sample for the presence of a GPCR gene nucleic acid, comprising in separate containers: a) one or more labeled nucleic acids comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the nucleotide sequence of said GPCR gene nucleic acid, and b) reagents for detection of said label.

48. The reagent kit of Claim 47, wherein the labeled nucleic acid comprises a contiguous nucleotide sequences which is completely complementary to a part of the nucleotide sequence of said GPCR gene nucleic acid.

49. A reagent kit for assaying a sample for the presence of a GPCR gene nucleic acid, comprising one or more nucleic acids comprising a contiguous nucleotide sequence which is at least partially complementary to a part of the nucleotide sequence of said GPCR gene nucleic acid, and which is capable of acting as a primer for said GPCR gene nucleic acid when maintained under conditions for primer extension.

50. The use of a nucleic acid which is 100 or fewer nucleotides in length and which is either: a) at least 80% identical to a contiguous sequence of nucleotides in one of the nucleic acid sequences as shown in Tables I and II; b) at least 80% identical to the complement of a contiguous sequence of nucleotides in one of the nucleic acid sequences as shown in Tables I and II; or c) capable of selectively hybridizing to said GPCR gene nucleic acid, for assaying a sample for the presence of a GPCR gene nucleic acid.

51. The use of a first nucleic acid which is 100 or fewer nucleotides in length and which is either: a) at least 80% identical to a contiguous sequence of nucleotides in one of the nucleic acid sequences as shown in Tables I and JJ; b) at least 80% identical to the complement of a contiguous sequence of nucleotides in one of the nucleic acid sequences as shown in Tables I and JJ; or c) capable of selectively hybridizing to said GPCR gene nucleic acid; for assaying a sample for the presence of a GPCR gene nucleic acid that has at least one nucleotide difference from the first nucleic acid.

52. The use of a nucleic acid which is 100 or fewer nucleotides in length and which is either: a) at least 80% identical to a contiguous sequence of nucleotides in one of the nucleic acid sequences as shown in Tables I and II; b) at least 80% identical to the complement of a contiguous sequence of nucleotides in one of the nucleic acid sequences as shown in Tables I and II; or c) capable of selectively hybridizing to said GPCR gene nucleic acid; for diagnosing a susceptibility to a disease or condition associated with a

GPCR.