CN113699160B - Mutation method of rat mitochondrial gene G14098A and application thereof - Google Patents
Mutation method of rat mitochondrial gene G14098A and application thereof Download PDFInfo
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- CN113699160B CN113699160B CN202110937125.7A CN202110937125A CN113699160B CN 113699160 B CN113699160 B CN 113699160B CN 202110937125 A CN202110937125 A CN 202110937125A CN 113699160 B CN113699160 B CN 113699160B
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- 230000035772 mutation Effects 0.000 title claims abstract description 63
- 108020005196 Mitochondrial DNA Proteins 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 17
- 238000011552 rat model Methods 0.000 claims abstract description 5
- 235000013601 eggs Nutrition 0.000 claims abstract description 4
- 125000006850 spacer group Chemical group 0.000 claims description 4
- 238000011144 upstream manufacturing Methods 0.000 claims description 3
- 238000010276 construction Methods 0.000 claims description 2
- 241000700159 Rattus Species 0.000 abstract description 79
- 230000002438 mitochondrial effect Effects 0.000 abstract description 8
- 238000004458 analytical method Methods 0.000 abstract description 7
- 201000010099 disease Diseases 0.000 abstract description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 7
- 230000004217 heart function Effects 0.000 abstract description 7
- 230000003542 behavioural effect Effects 0.000 abstract description 4
- 238000011156 evaluation Methods 0.000 abstract description 4
- 238000000520 microinjection Methods 0.000 abstract description 4
- 238000013461 design Methods 0.000 abstract description 3
- 230000009437 off-target effect Effects 0.000 abstract description 2
- 238000012216 screening Methods 0.000 abstract 1
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Abstract
大鼠线粒体基因G14098A的突变方法及其应用。本发明利用DdCBE碱基编辑技术，依据目标突变位点，设计两条包含突变位点的TALE靶点识别序列，通过筛选DdCBE组合，将DdCBE定位到相应的目标序列以使目标碱基由G突变为A。本发明的突变方法具有高精确性和低脱靶效应。本发明通过大鼠受精卵显微注射，能够实现大鼠的线粒体基因第14098位的G突变为A。雌性突变大鼠与野生型大鼠杂交进行突变传代分析，表明突变能够稳定传代；行为学和心脏功能评价证实该大鼠模型具有人类线粒体突变疾病表型。
Mutation method and application of rat mitochondrial gene G14098A. The present invention uses DdCBE base editing technology to design two TALE target recognition sequences containing the mutation site according to the target mutation site, and locates the DdCBE to the corresponding target sequence by screening the DdCBE combination so that the target base is mutated from G for A. The mutation method of the present invention has high precision and low off-target effects. The present invention can realize the mutation of G at the 14098th position of the mitochondrial gene of rats to A through microinjection of fertilized eggs of rats. Mutant passage analysis of female mutant rats crossed with wild-type rats showed that the mutations could be passed down stably; behavioral and cardiac function evaluations confirmed that the rat model had the phenotype of human mitochondrial mutation disease.
Description
技术领域technical field
本发明涉及一种线粒体基因突变方法及其动物建模应用。The invention relates to a mitochondrial gene mutation method and its application in animal modeling.
背景技术Background technique
随着生命科学和医学研究迈入新领域，对动物模型的需求正在日益上升。动物模型是疫苗、药物研发过程中不可逾越的环节，特别是重大传染病和复杂人类疾病的研究，都需要稳定有效的疾病动物模型作为支撑。As life science and medical research enters new territories, the demand for animal models is increasing. Animal models are an insurmountable link in the process of vaccine and drug development, especially the research on major infectious diseases and complex human diseases requires stable and effective disease animal models as support.
线粒体DNA(mtDNA)突变主要表现为碱基变化，它能够导致人类发生多种系统性疾病。迄今为止，人类mtDNA的致病变异已超过270种，并且数量还在不断上升。目前，仍没有针对mtDNA致病性突变的有效治疗方案。因此，迫切需求包含精确的人类mtDNA变异的动物模型来揭示这类疾病的病理生理学过程并开发针对这些疾病的治疗方法。此前，具有致病性mtDNA突变的哺乳动物动物模型可以通过线粒体移植获得或者使用PolGD257A/WT谱系筛选新的mtDNA突变[Stewart,J.B.J Inherit Metab Dis,2020]。然而，这两种策略都不能根据需求得到具有精确mtDNA突变的动物模型。Mitochondrial DNA (mtDNA) mutations are mainly manifested as base changes, which can lead to a variety of systemic diseases in humans. So far, there are more than 270 pathogenic variants of human mtDNA, and the number is still rising. Currently, there is still no effective treatment for mtDNA pathogenic mutations. Therefore, animal models containing accurate human mtDNA variation are urgently needed to reveal the pathophysiology of such diseases and develop therapeutic methods against these diseases. Previously, mammalian animal models with pathogenic mtDNA mutations could be obtained by mitochondrial transplantation or screened for novel mtDNA mutations using the PolG D257A/WT lineage [Stewart, JBJ Inherit Metab Dis, 2020]. However, neither of these strategies yields animal models with precise mtDNA mutations on demand.
发明内容Contents of the invention
在人体中，mtDNA TRNE基因中的G14710A突变与线粒体肌病相关，发明人发现其在大鼠中对应的位点是G14098，并因此设计了一种基因突变方法来应用于G14098A突变大鼠建模以辅助研究相关疾病。In humans, the G14710A mutation in the mtDNA TRNE gene is associated with mitochondrial myopathy. The inventors found that its corresponding site in rats is G14098, and therefore designed a gene mutation method to apply the G14098A mutant rat model To assist in the research of related diseases.
本发明的目的因此是提供一种精准有效的大鼠线粒体基因G14098A的突变方法及其相关应用。Therefore, the object of the present invention is to provide a precise and effective mutation method of rat mitochondrial gene G14098A and related applications.
根据本发明的第一方面，提供了一种大鼠线粒体基因G14098A的突变方法，包括：According to the first aspect of the present invention, a method for mutating rat mitochondrial gene G14098A is provided, comprising:
在目标突变碱基G位置上下游分别选定一个TALE识别靶序列，两个TALE识别靶序列中间的间隔序列为7-18bp且不含除目标突变碱基G之外的碱基G，其中两个TALE识别靶序列与间隔序列一起限定目标突变序列；A TALE recognition target sequence is selected at the upstream and downstream of the target mutation base G position, and the spacer sequence between the two TALE recognition target sequences is 7-18bp and does not contain base G other than the target mutation base G. A TALE recognition target sequence together with a spacer sequence defines a target mutation sequence;
根据选定的TALE识别靶序列筛选一对DdCBE组合；以及Screen a pair of DdCBE combinations according to the selected TALE recognition target sequence; and
共注射所筛选的DdCBE组合以将其定位于目标突变序列，从而将目标突变碱基G编辑为A而引入突变。The screened DdCBE combination was co-injected to locate it at the target mutation sequence, thereby editing the target mutation base G to A to introduce mutations.
本发明采用由DddA衍生的胞嘧啶碱基编辑器(DdCBE)成功实现了mtDNA中C·G到T·A的精准转变(高精确性和低脱靶效应)，从而为建立具有特异性mtDNA突变的动物模型以及相关研究应用开辟了途径。The present invention uses the cytosine base editor (DdCBE) derived from DddA to successfully realize the precise transformation of C·G to T·A in mtDNA (high accuracy and low off-target effect), so as to establish a specific mtDNA mutation Animal models and related research applications open avenues.
根据本发明的优选方案，所选定的两个TALE识别靶序列可以分别为：According to the preferred scheme of the present invention, the two selected TALE recognition target sequences can be respectively:
左侧识别靶序列：ttaactgtgactaat(SEQ ID NO.1)；以及Left recognition target sequence: ttaactgtgactaat (SEQ ID NO.1); and
右侧识别靶序列：agttgaattacggcgat(SEQ ID NO.2)。Target sequence recognized on the right: agttgaattacggcgat (SEQ ID NO. 2).
根据本发明的进一步优选方案，所筛选的DdCBE组合可以为：According to a further preferred solution of the present invention, the screened DdCBE combination can be:
Rat G14098A Left TALE-G1333C(L1333C)和Rat G14098A Right TALE-G1333N(R1333N)；Rat G14098A Left TALE-G1333C(L1333C) and Rat G14098A Right TALE-G1333N(R1333N);
Rat G14098A Left TALE-G1397C(L1397C)和Rat G14098A Right TALE-G1397N(L1397N)；Rat G14098A Left TALE-G1397C(L1397C) and Rat G14098A Right TALE-G1397N(L1397N);
Rat G14098A Left TALE-G1333N(L1333N)和Rat G14098A Right TALE-G1397C(R1333C)；或Rat G14098A Left TALE-G1333N(L1333N) and Rat G14098A Right TALE-G1397C(R1333C); or
Rat G14098A Left TALE-G1397N(L1397N)或Rat G14098A Right TALE-G1333C(L1397C)。Rat G14098A Left TALE-G1397N(L1397N) or Rat G14098A Right TALE-G1333C(L1397C).
DdCBE组合更优选为：Rat G14098A Left TALE-G1397C(L1397C)和Rat G14098ARight TALE-G1397N(L1397N)。The DdCBE combination is more preferably: Rat G14098A Left TALE-G1397C (L1397C) and Rat G14098ARight TALE-G1397N (L1397N).
根据本发明的另外方面，还提供了上述方法在构建线粒体基因靶位点G14098A突变大鼠模型中的应用。According to another aspect of the present invention, it also provides the application of the above method in the construction of a mitochondrial gene target site G14098A mutant rat model.
通过本发明的方法所建立的F0代线粒体基因G14098A雌性突变大鼠与野生型大鼠交配，在子代大鼠中能够检测到线粒体基因G14098A的突变。另外，通过对这种线粒体基因G14098A突变大鼠进行行为学和心脏功能分析，还能够模拟研究人线粒体基因G14710A突变的临床表型。The F0 generation of mitochondrial gene G14098A mutant rats established by the method of the present invention is mated with wild-type rats, and the mutation of mitochondrial gene G14098A can be detected in the offspring rats. In addition, by analyzing the behavior and heart function of the mitochondrial gene G14098A mutant rats, the clinical phenotype of the human mitochondrial gene G14710A mutation can also be simulated.
根据本发明的其它方面，还提供了上述方法在模拟研究人类线粒体G14710A突变所造成的线粒体脑肌病中的应用。According to other aspects of the present invention, it also provides the application of the above method in the simulated study of mitochondrial encephalomyopathy caused by human mitochondrial G14710A mutation.
附图说明Description of drawings
图1人和大鼠线粒体致病突变位点同源比对；Figure 1 Homologous comparison of pathogenic mutation sites in human and rat mitochondria;
图2在C6细胞中转染不同DdCBE组合产生线粒体G14098A突变比较；Figure 2 Comparison of the mitochondrial G14098A mutation produced by transfection of different DdCBE combinations in C6 cells;
图3线粒体基因G14098A突变大鼠鉴定结果；Fig. 3 Identification results of mitochondrial gene G14098A mutant rats;
图4线粒体基因G14098A突变大鼠各个组织突变情况分析；Figure 4 Analysis of mutations in various tissues of mitochondrial gene G14098A mutant rats;
图5线粒体基因G14098A突变大鼠相关行为分析；Figure 5 Analysis of related behaviors in mitochondrial gene G14098A mutant rats;
图6线粒体基因G14098A突变大鼠心脏功能检测。Fig. 6 Detection of cardiac function in rats with mitochondrial gene G14098A mutation.
具体实施方式Detailed ways
首先，根据人的线粒体基因突变G14710A，确定对应大鼠线粒体基因突变G14098A(见图1)。根据大鼠的突变位点，设计TALE的识别位点(靶点或靶序列)。本发明采用DdCBE定点线粒体编辑：利用一对TALE识别位点将融合的碱基编辑工具定位到靶点基因位置，从而实现目标碱基的突变。本发明如下选择设计TALE识别靶点：First, according to the human mitochondrial gene mutation G14710A, the corresponding rat mitochondrial gene mutation G14098A was determined (see FIG. 1 ). According to the mutation site of the rat, the recognition site (target site or target sequence) of the TALE is designed. The present invention uses DdCBE fixed-point mitochondrial editing: a pair of TALE recognition sites is used to locate the fused base editing tool to the position of the target gene, thereby realizing the mutation of the target base. The present invention selects and designs TALE recognition targets as follows:
根据TALE的靶点设计原则，在靶点上下游分别选定一个TALE识别位点，两个TALE识别位点中间间隔序列为7-18bp，其包含目标突变碱基G，同时避免目标碱基外的碱基G。According to the principle of TALE target design, a TALE recognition site is selected in the upstream and downstream of the target respectively. base G.
利用两个融合TALE识别位点的DdCBE定位于相应的目标突变序列，并使目标突变G编辑变为A，从而在大鼠线粒体基因中引入突变。The DdCBE using two fused TALE recognition sites was positioned at the corresponding target mutation sequence, and the target mutation G was edited into A, thereby introducing mutations in the rat mitochondrial gene.
针对大鼠线粒体基因G14098突变，本发明选定了如下两条TALE左右识别靶点：For the mutation of rat mitochondrial gene G14098, the present invention selects the following two TALE recognition targets:
左侧识别靶点：Identify the target on the left:
ttaactgtgactaat(SEQ ID NO.1)ttaactgtgactaat (SEQ ID NO.1)
右侧识别靶点：Identify the target on the right:
agttgaattacggcgat(SEQ ID NO.2)agttgaattacggcgat (SEQ ID NO.2)
针对上述选定的TALE靶点序列，构建不同的DdCBE，对应不同的DdCBE如下：For the above selected TALE target sequences, construct different DdCBEs, corresponding to different DdCBEs as follows:
Rat G14098A Left TALE-G1333C(SEQ ID NO.3)；Rat G14098A Left TALE-G1333C (SEQ ID NO.3);
Rat G14098A Right TALE-G1333N(SEQ ID NO.4)；Rat G14098A Right TALE-G1333N (SEQ ID NO.4);
Rat G14098A Left TALE-G1397C(SEQ ID NO.5)；Rat G14098A Left TALE-G1397C (SEQ ID NO.5);
Rat G14098A Right TALE-G1397N(SEQ ID NO.6)；Rat G14098A Right TALE-G1397N (SEQ ID NO.6);
Rat G14098A Left TALE-G1333N(SEQ ID NO.7)；Rat G14098A Left TALE-G1333N (SEQ ID NO.7);
Rat G14098A Right TALE-G1397C(SEQ ID NO.8)；Rat G14098A Right TALE-G1397C (SEQ ID NO.8);
Rat G14098A Left TALE-G1397N(SEQ ID NO.9)；Rat G14098A Left TALE-G1397N (SEQ ID NO.9);
Rat G14098A Right TALE-G1333C(SEQ ID NO.10)。Rat G14098A Right TALE-G1333C (SEQ ID NO. 10).
将DdCBE分别导入PB转座子表达载体，用于大鼠线粒体基因G14098A的突变。DdCBEs were introduced into PB transposon expression vectors for the mutation of rat mitochondrial gene G14098A.
实施例1Example 1
在大鼠细胞系上进行DdCBE介导的线粒体基因G14098A突变。按常规操作，进行细胞系的基因编辑(通过电转或脂质体转染)，以脂质体转染为例。DdCBE-mediated mutation of the mitochondrial gene G14098A was performed on a rat cell line. Perform gene editing of cell lines (by electroporation or lipofection) according to routine operations, taking lipofection as an example.
(1)以C6细胞为例，本发明进行真核生物细胞的培养与转染：C6细胞接种培养于添加10％FBS(Gemini)的DMEM高糖培养液中,其中含penicillin(100U/ml)和streptomycin(100μg/ml)。(1) Taking C6 cells as an example, the present invention carries out the cultivation and transfection of eukaryotic cells: C6 cells are inoculated and cultured in DMEM high-sugar culture medium supplemented with 10% FBS (Gemini), which contains penicillin (100U/ml) and streptomycin (100 μg/ml).
(2)在转染前分至6孔板中，待密度达到70％-80％时进行转染。(2) Divide into 6-well plates before transfection, and perform transfection when the density reaches 70%-80%.
DdCBE系统质粒组合如下：The DdCBE system plasmid combination is as follows:
A:Rat G14098A Left TALE-G1333C/Rat G14098A Right TALE-G1333N(L1333C+R1333N)；A:Rat G14098A Left TALE-G1333C/Rat G14098A Right TALE-G1333N(L1333C+R1333N);
B:Rat G14098A Left TALE-G1397C/Rat G14098A Right TALE-G1397N(L1397C+R1397N)；B:Rat G14098A Left TALE-G1397C/Rat G14098A Right TALE-G1397N(L1397C+R1397N);
C:Rat G14098A Left TALE-G1333N/Rat G14098A Right TALE-G1397C(L1333N+RL1333C)；C:Rat G14098A Left TALE-G1333N/Rat G14098A Right TALE-G1397C(L1333N+RL1333C);
D:Rat G14098A Left TALE-G1397N/Rat G14098A Right TALE-G1333C(L1397N+RL1397C)；D:Rat G14098A Left TALE-G1397N/Rat G14098A Right TALE-G1333C(L1397N+RL1397C);
(3)转染以脂质体转染为例。按照LipofectamineTM2000Transfection Reagent(Invitrogen,11668-019)的操作手册，以A组合为例，将1μg Rat G14098A Left TALE-G1333C(L1333C)质粒与1μg Rat G14098A Right TALE-G1333N(R1333N)质粒混匀，共转染至每孔细胞中，6-8小时后换液，72小时后收取细胞。(3) Transfection Take liposome transfection as an example. According to the operation manual of Lipofectamine TM 2000Transfection Reagent (Invitrogen, 11668-019), taking combination A as an example, mix 1 μg Rat G14098A Left TALE-G1333C (L1333C) plasmid with 1 μg Rat G14098A Right TALE-G1333N (R1333N) plasmid, and mix together Transfect into the cells in each well, change the medium after 6-8 hours, and harvest the cells after 72 hours.
(4)基因型分析(4) Genotype analysis
A、收取部分细胞在裂解液(10μM Tris-HCl,0.4M NaCl,2μMEDTA,1％SDS)中用100μg/ml蛋白酶K裂解消化后，酚-氯仿抽提后溶解到50μl去离子水中。A. Harvest part of the cells, lyse and digest them with 100 μg/ml proteinase K in lysate (10 μM Tris-HCl, 0.4M NaCl, 2 μM EDTA, 1% SDS), dissolve them in 50 μl deionized water after extraction with phenol-chloroform.
B、使用一对引物G14098-For(TCAAGTCTCCGGGTACTCCT)和G14098-Rev(AAATATTGAGGCGCCGTTGG)进行PCR扩增，用AxyPrep PCR cleanup试剂盒(AXYGEN，AP-PCR-250G)纯化获得PCR回收产物，PCR反应体系为：B. Use a pair of primers G14098-For (TCAAGTCTCCGGGTACTCCT) and G14098-Rev (AAATATTGAGGCGCCGTTGG) for PCR amplification, and use AxyPrep PCR cleanup kit (AXYGEN, AP-PCR-250G) to purify and obtain PCR recovered products. The PCR reaction system is:
300-400ng基因组DNA300-400ng genomic DNA
25μl 2×Buffer25μl 2×Buffer
1μl dNTP1 μl dNTPs
2μl G14098--For(20μm)2μl G14098--For (20μm)
2μl G14098--Rev(20μm)2μl G14098--Rev (20μm)
1μl DNA Polymerase(Vazyme,P505-d3)1μl DNA Polymerase (Vazyme, P505-d3)
补水至50μl体系。Replenish water to 50μl system.
C、获得的PCR回收产物连接到克隆载体
1μl PCR产物1 μl PCR product
1μl
轻轻混合，室温(20-37℃)反应5分钟。反应结束后，将离心管置于冰上。Mix gently and react at room temperature (20-37°C) for 5 minutes. After the reaction, place the centrifuge tube on ice.
连接产物转化DH5感受态细胞(TransGen,CD201)。The ligation product was transformed into DH5 competent cells (TransGen, CD201).
D、挑取克隆，用通用引物M13-F测序靶基因突变，测序结果如下(斜体、加粗、下划线表示突变碱基)：D. Pick clones and use the universal primer M13-F to sequence the mutation of the target gene. The sequencing results are as follows (italics, bold, and underline indicate the mutated bases):
靶点突变情况：Target Mutations:
Wt：gacatgaaaaaWt: gacatgaaaaa
Mut：gacataaaaaaMut: gacat a aaaaa
结果表明：线粒体基因14098位点由G突变为了A，并且DdCBE组合B：G14098A LeftTALE-G1397C/Rat G14098A Right TALE-G1397N介导的线粒体G14098A突变效率最高(图2)。The results showed that the mitochondrial gene 14098 site was mutated from G to A, and DdCBE combination B: G14098A LeftTALE-G1397C/Rat G14098A Right TALE-G1397N mediated the highest mitochondrial G14098A mutation efficiency (Figure 2).
实施例2Example 2
在大鼠受精卵上进行DdCBE介导的线粒体基因G14098A突变，实现目标碱基突变，建立线粒体基因G14098突变大鼠模型，并进行行为学和心脏功能评价，鉴定该模型是否符合人的线粒体突变导致的临床表型。DdCBE-mediated mitochondrial gene G14098A mutation was carried out on fertilized rat eggs to achieve the target base mutation, and the mitochondrial gene G14098 mutant rat model was established, and behavioral and cardiac function evaluations were performed to identify whether the model was consistent with human mitochondrial mutations. clinical phenotype.
按常规操作进行大鼠的胚胎收集、显微注射、胚胎培养和胚胎移植等。Rat embryo collection, microinjection, embryo culture and embryo transfer were performed according to routine operations.
(1)显微注射：受精卵注射G14098A Left TALE-G1397C/Rat G14098A RightTALE-G1397N质粒。常规进行胚胎移植；(1) Microinjection: the fertilized eggs were injected with the G14098A Left TALE-G1397C/Rat G14098A RightTALE-G1397N plasmid. Routine embryo transfer;
(2)基因型分析：(2) Genotype analysis:
A、常规小鼠剪尾提取基因组DNA：在裂解液(10μM Tris-HCl,0.4M NaCl,2μMEDTA,1％SDS)中用100μg/ml蛋白酶K裂解消化后，酚-氯仿抽提后溶解到50μl去离子水中。A. Genomic DNA was extracted by pruning the tail of conventional mice: lysed and digested with 100 μg/ml proteinase K in the lysate (10 μM Tris-HCl, 0.4M NaCl, 2 μM EDTA, 1% SDS), and dissolved to 50 μl after extraction with phenol-chloroform deionized water.
B、使用一对引物G14098-For(TCAAGTCTCCGGGTACTCCT)和G14098-Rev(AAATATTGAGGCGCCGTTGG)进行PCR扩增，用AxyPrep PCR cleanup试剂盒(AXYGEN，AP-PCR-250G)纯化获得PCR回收产物，PCR反应体系为：B. Use a pair of primers G14098-For (TCAAGTCTCCGGGTACTCCT) and G14098-Rev (AAATATTGAGGCGCCGTTGG) for PCR amplification, and use AxyPrep PCR cleanup kit (AXYGEN, AP-PCR-250G) to purify and obtain PCR recovered products. The PCR reaction system is:
300-400ng基因组DNA300-400ng genomic DNA
25μl 2×Buffer25μl 2×Buffer
1μl dNTP1 μl dNTPs
2μl G14098--For(20μm)2μl G14098--For (20μm)
2μl G14098--Rev(20μm)2μl G14098--Rev (20μm)
1μl DNA Polymerase(Vazyme,P505-d3)1μl DNA Polymerase (Vazyme, P505-d3)
补水至50μl体系。Replenish water to 50μl system.
C、获得的PCR回收产物连接到克隆载体
2μl PCR产物2 μl PCR product
1μl
轻轻混合，室温(20-37℃)反应5分钟。反应结束后，将离心管置于冰上。Mix gently and react at room temperature (20-37°C) for 5 minutes. After the reaction, place the centrifuge tube on ice.
连接产物转化DH5感受态细胞(TransGen,CD201)。The ligation product was transformed into DH5 competent cells (TransGen, CD201).
D、挑取克隆，用通用引物M13-F测序靶基因突变，测序结果如下(斜体加粗下划表示突变碱基)：D. Pick the clones and use the universal primer M13-F to sequence the mutation of the target gene. The sequencing results are as follows (italics, bold and underlined indicate the mutated bases):
靶点突变情况：Target Mutations:
Wt：gacatgaaaaaWt: gacatgaaaaa
Mut：gacataaaaaaMut: gacataaaaaa
结果表明：线粒体基因14098位点由G突变为了A(见图3)，并且我们取材一只突变大鼠进行了各个组织的突变情况分析，结果显示各个组织都能够检测到突变(见图3)，结果证实利用G14098A Left TALE-G1397C/Rat G14098A Right TALE-G1397N质粒，通过显微注射的方式，可以实现线粒体基因G14098A的突变。The results showed that the 14098 site of the mitochondrial gene was mutated from G to A (see Figure 3), and we took a mutant rat and analyzed the mutations in various tissues, and the results showed that mutations could be detected in all tissues (see Figure 3) , the results confirmed that the mitochondrial gene G14098A mutation can be achieved by microinjection using the G14098A Left TALE-G1397C/Rat G14098A Right TALE-G1397N plasmid.
(4)通过将获得的G1408A雌雄突变大鼠与野生型大鼠进行杂交，对子代大鼠进行基因组DNA提取、PCR和测序分析，发现突变能够稳定的遗传到子代大鼠中(见图4)。表明我们获得了能够稳定遗传的线粒体基因G14098A突变大鼠。(4) By crossing the obtained G1408A male and female mutant rats with wild-type rats, the offspring rats were subjected to genomic DNA extraction, PCR and sequencing analysis, and it was found that the mutation could be stably inherited to the offspring rats (see figure 4). It shows that we have obtained mitochondrial gene G14098A mutant rats that can be stably inherited.
(5)线粒体基因G14098A点突变大鼠行为学分析(5) Behavioral analysis of mitochondrial gene G14098A point mutation rats
获得的线粒体基因G14098A点突变大鼠，会导致线粒体功能缺陷，进而影响肌肉组织的功能，通过行为学方法检测，判定大鼠的肌肉功能指标。The obtained mitochondrial gene G14098A point mutation rats will lead to mitochondrial functional defects, which will affect the function of muscle tissue. The muscle function indicators of the rats are determined by behavioral methods.
开放旷场实验：该实验在80cm×80cm×50cm的黑色箱体中进行。使用SuperMaze数字跟踪系统记录大鼠自发活动5分钟。大鼠的运动轨迹通过SuperMaze软件进行分析。Open field experiment: This experiment is carried out in a black box of 80cm×80cm×50cm. Rat locomotor activity was recorded for 5 min using the SuperMaze digital tracking system. The trajectories of the rats were analyzed by SuperMaze software.
转棒实验：使用带有自动计时器和跌落传感器的旋转棒仪(ZH-600B，安徽正华生物仪器设备公司)评估大鼠的运动及平衡能力。训练前，所有大鼠均在固定转棒上停留3分钟。采用20rpm的速度进行训练，大鼠跌落后立即放回转棒上，最多放回5次。该测试每天进行一次，连续三天。记录大鼠掉落转棒的潜伏期。Rotarod test: A rotarod instrument (ZH-600B, Anhui Zhenghua Bio-Instrument Co., Ltd.) with an automatic timer and a drop sensor was used to evaluate the movement and balance ability of rats. Before training, all rats stayed on a fixed rotarod for 3 minutes. Adopt the speed of 20rpm to carry out training, rat is put back on the rotating bar immediately after falling, put back 5 times at most. The test is performed once a day for three consecutive days. The latency period for rats to drop the rotarod was recorded.
抓力实验：使用握力计(ZH-YLS-13A，安徽正华生物设备设施)测量大鼠四肢的肌肉力量。记录三次测量值，计算平均值。Grip force test: The muscle strength of the rat limbs was measured using a grip dynamometer (ZH-YLS-13A, Anhui Zhenghua Bio-equipment Facility). Three measurements were recorded and the average value calculated.
开放旷场的实验结果显示突变导致大鼠运动距离和平均速度下降，运动和平衡能力下降，四肢抓力显著降低，如图5所示。The experimental results in the open field showed that the mutation caused the rats to decrease the movement distance and average speed, the movement and balance ability, and the grasping force of the limbs were significantly reduced, as shown in Figure 5.
(6)线粒体基因G14098A突变大鼠心脏功能评价(6) Cardiac function evaluation of mitochondrial gene G14098A mutant rats
获得的线粒体基因G14098A突变大鼠，用超声心动图分析检测心脏功能相关的参数。The obtained mitochondrial gene G14098A mutant rats were analyzed by echocardiography to detect parameters related to cardiac function.
心动超声法：用异氟醚麻醉大鼠，并使用微超声成像系统(Vevo3100)进行超声心动图观察。测量记录至少三个连续心动周期。Cardiac ultrasonography: Rats were anesthetized with isoflurane, and echocardiographic observation was performed using a micro-ultrasound imaging system (Vevo3100). Measurements were recorded for at least three consecutive cardiac cycles.
心脏功能评价结果显示与野生大鼠相比，突变大鼠表现出扩张型心肌病表型，心室更大，壁更薄，收缩功能下降，如图6所示。Cardiac function evaluation results showed that compared with wild rats, mutant rats exhibited dilated cardiomyopathy phenotype, with larger ventricles, thinner walls, and decreased systolic function, as shown in Figure 6.
序列表sequence listing
<110> 中国医学科学院医学实验动物研究所<110> Institute of Medical Experimental Animals, Chinese Academy of Medical Sciences
<120> 大鼠线粒体基因G14098A的突变方法及其应用<120> Mutation method and application of rat mitochondrial gene G14098A
<160> 10<160> 10
<170> SIPOSequenceListing 1.0<170> SIPOSequenceListing 1.0
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<213> Artificial Sequence<213> Artificial Sequence
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ttaactgtga ctaat 15
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<213> Artificial Sequence<213> Artificial Sequence
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Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly AlaMet Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 151 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu LeuLeu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30 20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala AlaLeu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45 35 40 45
Gln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala AspGln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala Asp
50 55 60 50 55 60
Tyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu GlyTyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu Gly
65 70 75 8065 70 75 80
Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser ThrTyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr
85 90 95 85 90 95
Val Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His AlaVal Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala
100 105 110 100 105 110
His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val AlaHis Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala
115 120 125 115 120 125
Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His GluVal Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu
130 135 140 130 135 140
Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu GluAla Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu
145 150 155 160145 150 155 160
Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln LeuAla Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu
165 170 175 165 170 175
Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr AlaAsp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala
180 185 190 180 185 190
Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro LeuVal Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
195 200 205 195 200 205
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyAsn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
210 215 220 210 215 220
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
225 230 235 240225 230 235 240
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn GlyAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
245 250 255 245 250 255
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
260 265 270 260 265 270
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
275 280 285 275 280 285
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProAsn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
290 295 300 290 295 300
Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
305 310 315 320305 310 315 320
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
325 330 335 325 330 335
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
340 345 350 340 345 350
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
355 360 365 355 360 365
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln
370 375 380 370 375 380
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
385 390 395 400385 390 395 400
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
405 410 415 405 410 415
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala LeuGlu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
420 425 430 420 425 430
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
435 440 445 435 440 445
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys GlnThr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
450 455 460 450 455 460
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
465 470 475 480465 470 475 480
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly GlyGly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
485 490 495 485 490 495
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
500 505 510 500 505 510
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn IleAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
515 520 525 515 520 525
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
530 535 540 530 535 540
Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser
545 550 555 560545 550 555 560
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProHis Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
565 570 575 565 570 575
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
580 585 590 580 585 590
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
595 600 605 595 600 605
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
610 615 620 610 615 620
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
625 630 635 640625 630 635 640
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
645 650 655 645 650 655
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
660 665 670 660 665 670
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
675 680 685 675 680 685
Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala LeuAsp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu
690 695 700 690 695 700
Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala AlaGlu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala
705 710 715 720705 710 715 720
Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg ProLeu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg Pro
725 730 735 725 730 735
Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Pro Thr Pro TyrAla Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Pro Thr Pro Tyr
740 745 750 740 745 750
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu PhePro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
755 760 765 755 760 765
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn ProMet Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
770 775 780 770 775 780
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu ProGlu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
785 790 795 800785 790 795 800
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro ValGlu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
805 810 815 805 810 815
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser AsnLys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
820 825 830 820 825 830
Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Gly Ser Thr AsnSer Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Gly Ser Thr Asn
835 840 845 835 840 845
Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile GlnLeu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln
850 855 860 850 855 860
Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly AsnGlu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn
865 870 875 880865 870 875 880
Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser ThrLys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr
885 890 895 885 890 895
Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys ProAsp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro
900 905 910 900 905 910
Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys MetTrp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met
915 920 925 915 920 925
LeuLeu
<210> 4<210> 4
<211> 947<211> 947
<212> PRT<212> PRT
<213> Artificial Sequence<213> Artificial Sequence
<400> 4<400> 4
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly AlaMet Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 151 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu LeuLeu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30 20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala AlaLeu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45 35 40 45
Gln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala AspGln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala Asp
50 55 60 50 55 60
Tyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu GlyTyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu Gly
65 70 75 8065 70 75 80
Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser ThrTyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr
85 90 95 85 90 95
Val Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His AlaVal Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala
100 105 110 100 105 110
His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val AlaHis Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala
115 120 125 115 120 125
Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His GluVal Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu
130 135 140 130 135 140
Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu GluAla Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu
145 150 155 160145 150 155 160
Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln LeuAla Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu
165 170 175 165 170 175
Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr AlaAsp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala
180 185 190 180 185 190
Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro LeuVal Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
195 200 205 195 200 205
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly GlyAsn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
210 215 220 210 215 220
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
225 230 235 240225 230 235 240
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn AsnAla His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
245 250 255 245 250 255
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
260 265 270 260 265 270
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
275 280 285 275 280 285
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProAsn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
290 295 300 290 295 300
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
305 310 315 320305 310 315 320
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
325 330 335 325 330 335
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
340 345 350 340 345 350
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
355 360 365 355 360 365
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
370 375 380 370 375 380
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
385 390 395 400385 390 395 400
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
405 410 415 405 410 415
Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala LeuAsp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
420 425 430 420 425 430
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
435 440 445 435 440 445
Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys GlnThr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
450 455 460 450 455 460
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
465 470 475 480465 470 475 480
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyGly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
485 490 495 485 490 495
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
500 505 510 500 505 510
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn IleAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
515 520 525 515 520 525
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
530 535 540 530 535 540
Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser
545 550 555 560545 550 555 560
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProHis Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
565 570 575 565 570 575
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
580 585 590 580 585 590
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
595 600 605 595 600 605
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
610 615 620 610 615 620
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
625 630 635 640625 630 635 640
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
645 650 655 645 650 655
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
660 665 670 660 665 670
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
675 680 685 675 680 685
Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala LeuAla Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
690 695 700 690 695 700
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
705 710 715 720705 710 715 720
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys GlnThr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
725 730 735 725 730 735
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
740 745 750 740 745 750
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyGly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
755 760 765 755 760 765
Arg Pro Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp ProArg Pro Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
770 775 780 770 775 780
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys LeuAla Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
785 790 795 800785 790 795 800
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly SerGly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
805 810 815 805 810 815
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu ProGly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
820 825 830 820 825 830
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp AlaAla Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
835 840 845 835 840 845
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Ser Gly Gly SerGly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Ser Gly Gly Ser
850 855 860 850 855 860
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu ValThr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
865 870 875 880865 870 875 880
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val IleIle Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
885 890 895 885 890 895
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp GluGly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
900 905 910 900 905 910
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu TyrSer Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
915 920 925 915 920 925
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys IleLys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
930 935 940 930 935 940
Lys Met LeuLys Met Leu
945945
<210> 5<210> 5
<211> 865<211> 865
<212> PRT<212> PRT
<213> Artificial Sequence<213> Artificial Sequence
<400> 5<400> 5
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly AlaMet Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 151 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu LeuLeu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30 20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala AlaLeu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45 35 40 45
Gln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala AspGln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala Asp
50 55 60 50 55 60
Tyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu GlyTyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu Gly
65 70 75 8065 70 75 80
Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser ThrTyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr
85 90 95 85 90 95
Val Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His AlaVal Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala
100 105 110 100 105 110
His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val AlaHis Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala
115 120 125 115 120 125
Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His GluVal Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu
130 135 140 130 135 140
Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu GluAla Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu
145 150 155 160145 150 155 160
Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln LeuAla Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu
165 170 175 165 170 175
Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr AlaAsp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala
180 185 190 180 185 190
Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro LeuVal Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
195 200 205 195 200 205
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyAsn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
210 215 220 210 215 220
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
225 230 235 240225 230 235 240
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn GlyAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
245 250 255 245 250 255
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
260 265 270 260 265 270
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
275 280 285 275 280 285
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProAsn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
290 295 300 290 295 300
Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
305 310 315 320305 310 315 320
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
325 330 335 325 330 335
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
340 345 350 340 345 350
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
355 360 365 355 360 365
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln
370 375 380 370 375 380
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
385 390 395 400385 390 395 400
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
405 410 415 405 410 415
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala LeuGlu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
420 425 430 420 425 430
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
435 440 445 435 440 445
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys GlnThr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
450 455 460 450 455 460
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
465 470 475 480465 470 475 480
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly GlyGly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
485 490 495 485 490 495
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
500 505 510 500 505 510
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn IleAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
515 520 525 515 520 525
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
530 535 540 530 535 540
Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser
545 550 555 560545 550 555 560
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProHis Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
565 570 575 565 570 575
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
580 585 590 580 585 590
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
595 600 605 595 600 605
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
610 615 620 610 615 620
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
625 630 635 640625 630 635 640
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
645 650 655 645 650 655
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
660 665 670 660 665 670
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
675 680 685 675 680 685
Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala LeuAsp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu
690 695 700 690 695 700
Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala AlaGlu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala
705 710 715 720705 710 715 720
Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg ProLeu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg Pro
725 730 735 725 730 735
Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Ala Ile Pro ValAla Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Ala Ile Pro Val
740 745 750 740 745 750
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser AsnLys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
755 760 765 755 760 765
Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Gly Ser Thr AsnSer Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Gly Ser Thr Asn
770 775 780 770 775 780
Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile GlnLeu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln
785 790 795 800785 790 795 800
Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly AsnGlu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn
805 810 815 805 810 815
Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser ThrLys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr
820 825 830 820 825 830
Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys ProAsp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro
835 840 845 835 840 845
Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys MetTrp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met
850 855 860 850 855 860
LeuLeu
865865
<210> 6<210> 6
<211> 1011<211> 1011
<212> PRT<212> PRT
<213> Artificial Sequence<213> Artificial Sequence
<400> 6<400> 6
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly AlaMet Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 151 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu LeuLeu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30 20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala AlaLeu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45 35 40 45
Gln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala AspGln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala Asp
50 55 60 50 55 60
Tyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu GlyTyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu Gly
65 70 75 8065 70 75 80
Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser ThrTyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr
85 90 95 85 90 95
Val Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His AlaVal Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala
100 105 110 100 105 110
His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val AlaHis Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala
115 120 125 115 120 125
Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His GluVal Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu
130 135 140 130 135 140
Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu GluAla Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu
145 150 155 160145 150 155 160
Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln LeuAla Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu
165 170 175 165 170 175
Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr AlaAsp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala
180 185 190 180 185 190
Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro LeuVal Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
195 200 205 195 200 205
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly GlyAsn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
210 215 220 210 215 220
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
225 230 235 240225 230 235 240
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn AsnAla His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
245 250 255 245 250 255
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
260 265 270 260 265 270
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
275 280 285 275 280 285
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProAsn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
290 295 300 290 295 300
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
305 310 315 320305 310 315 320
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
325 330 335 325 330 335
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
340 345 350 340 345 350
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
355 360 365 355 360 365
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
370 375 380 370 375 380
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
385 390 395 400385 390 395 400
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
405 410 415 405 410 415
Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala LeuAsp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
420 425 430 420 425 430
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
435 440 445 435 440 445
Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys GlnThr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
450 455 460 450 455 460
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
465 470 475 480465 470 475 480
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyGly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
485 490 495 485 490 495
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
500 505 510 500 505 510
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn IleAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
515 520 525 515 520 525
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
530 535 540 530 535 540
Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser
545 550 555 560545 550 555 560
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProHis Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
565 570 575 565 570 575
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
580 585 590 580 585 590
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
595 600 605 595 600 605
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
610 615 620 610 615 620
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
625 630 635 640625 630 635 640
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
645 650 655 645 650 655
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
660 665 670 660 665 670
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
675 680 685 675 680 685
Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala LeuAla Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
690 695 700 690 695 700
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
705 710 715 720705 710 715 720
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys GlnThr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
725 730 735 725 730 735
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
740 745 750 740 745 750
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyGly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
755 760 765 755 760 765
Arg Pro Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp ProArg Pro Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
770 775 780 770 775 780
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys LeuAla Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
785 790 795 800785 790 795 800
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly SerGly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
805 810 815 805 810 815
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu ProGly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
820 825 830 820 825 830
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp AlaAla Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
835 840 845 835 840 845
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro TyrGly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
850 855 860 850 855 860
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu PhePro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
865 870 875 880865 870 875 880
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn ProMet Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
885 890 895 885 890 895
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu ProGlu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
900 905 910 900 905 910
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ser Gly Gly SerGlu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ser Gly Gly Ser
915 920 925 915 920 925
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu ValThr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
930 935 940 930 935 940
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val IleIle Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
945 950 955 960945 950 955 960
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp GluGly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
965 970 975 965 970 975
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu TyrSer Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
980 985 990 980 985 990
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys IleLys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
995 1000 1005 995 1000 1005
Lys Met LeuLys Met Leu
1010 1010
<210> 7<210> 7
<211> 879<211> 879
<212> PRT<212> PRT
<213> Artificial Sequence<213> Artificial Sequence
<400> 7<400> 7
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly AlaMet Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 151 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu LeuLeu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30 20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala AlaLeu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45 35 40 45
Gln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala AspGln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala Asp
50 55 60 50 55 60
Tyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu GlyTyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu Gly
65 70 75 8065 70 75 80
Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser ThrTyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr
85 90 95 85 90 95
Val Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His AlaVal Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala
100 105 110 100 105 110
His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val AlaHis Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala
115 120 125 115 120 125
Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His GluVal Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu
130 135 140 130 135 140
Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu GluAla Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu
145 150 155 160145 150 155 160
Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln LeuAla Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu
165 170 175 165 170 175
Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr AlaAsp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala
180 185 190 180 185 190
Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro LeuVal Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
195 200 205 195 200 205
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyAsn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
210 215 220 210 215 220
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
225 230 235 240225 230 235 240
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn GlyAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
245 250 255 245 250 255
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
260 265 270 260 265 270
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
275 280 285 275 280 285
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProAsn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
290 295 300 290 295 300
Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
305 310 315 320305 310 315 320
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
325 330 335 325 330 335
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
340 345 350 340 345 350
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
355 360 365 355 360 365
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln
370 375 380 370 375 380
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
385 390 395 400385 390 395 400
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
405 410 415 405 410 415
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala LeuGlu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
420 425 430 420 425 430
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
435 440 445 435 440 445
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys GlnThr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
450 455 460 450 455 460
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
465 470 475 480465 470 475 480
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly GlyGly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
485 490 495 485 490 495
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
500 505 510 500 505 510
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn IleAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
515 520 525 515 520 525
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
530 535 540 530 535 540
Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser
545 550 555 560545 550 555 560
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProHis Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
565 570 575 565 570 575
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
580 585 590 580 585 590
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
595 600 605 595 600 605
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
610 615 620 610 615 620
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
625 630 635 640625 630 635 640
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
645 650 655 645 650 655
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
660 665 670 660 665 670
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
675 680 685 675 680 685
Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala LeuAsp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu
690 695 700 690 695 700
Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala AlaGlu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala
705 710 715 720705 710 715 720
Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg ProLeu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg Pro
725 730 735 725 730 735
Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Gly Ser Tyr AlaAla Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Gly Ser Tyr Ala
740 745 750 740 745 750
Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn GlyLeu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly
755 760 765 755 760 765
Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu GluGln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu
770 775 780 770 775 780
Ser Lys Val Phe Ser Ser Gly Gly Ser Gly Gly Ser Thr Asn Leu SerSer Lys Val Phe Ser Ser Gly Gly Ser Gly Gly Ser Thr Asn Leu Ser
785 790 795 800785 790 795 800
Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln Glu SerAsp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser
805 810 815 805 810 815
Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys ProIle Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro
820 825 830 820 825 830
Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp GluGlu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu
835 840 845 835 840 845
Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp AlaAsn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala
850 855 860 850 855 860
Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met LeuLeu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu
865 870 875865 870 875
<210> 8<210> 8
<211> 933<211> 933
<212> PRT<212> PRT
<213> Artificial Sequence<213> Artificial Sequence
<400> 8<400> 8
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly AlaMet Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 151 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu LeuLeu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30 20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala AlaLeu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45 35 40 45
Gln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala AspGln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala Asp
50 55 60 50 55 60
Tyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu GlyTyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu Gly
65 70 75 8065 70 75 80
Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser ThrTyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr
85 90 95 85 90 95
Val Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His AlaVal Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala
100 105 110 100 105 110
His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val AlaHis Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala
115 120 125 115 120 125
Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His GluVal Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu
130 135 140 130 135 140
Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu GluAla Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu
145 150 155 160145 150 155 160
Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln LeuAla Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu
165 170 175 165 170 175
Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr AlaAsp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala
180 185 190 180 185 190
Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro LeuVal Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
195 200 205 195 200 205
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly GlyAsn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
210 215 220 210 215 220
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
225 230 235 240225 230 235 240
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn AsnAla His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
245 250 255 245 250 255
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
260 265 270 260 265 270
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
275 280 285 275 280 285
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProAsn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
290 295 300 290 295 300
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
305 310 315 320305 310 315 320
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
325 330 335 325 330 335
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
340 345 350 340 345 350
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
355 360 365 355 360 365
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
370 375 380 370 375 380
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
385 390 395 400385 390 395 400
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
405 410 415 405 410 415
Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala LeuAsp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
420 425 430 420 425 430
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
435 440 445 435 440 445
Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys GlnThr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
450 455 460 450 455 460
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
465 470 475 480465 470 475 480
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyGly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
485 490 495 485 490 495
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
500 505 510 500 505 510
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn IleAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
515 520 525 515 520 525
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
530 535 540 530 535 540
Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser
545 550 555 560545 550 555 560
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProHis Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
565 570 575 565 570 575
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
580 585 590 580 585 590
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
595 600 605 595 600 605
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
610 615 620 610 615 620
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
625 630 635 640625 630 635 640
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
645 650 655 645 650 655
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
660 665 670 660 665 670
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
675 680 685 675 680 685
Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala LeuAla Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
690 695 700 690 695 700
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
705 710 715 720705 710 715 720
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys GlnThr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
725 730 735 725 730 735
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
740 745 750 740 745 750
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyGly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
755 760 765 755 760 765
Arg Pro Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp ProArg Pro Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
770 775 780 770 775 780
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys LeuAla Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
785 790 795 800785 790 795 800
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly SerGly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
805 810 815 805 810 815
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe ThrAla Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
820 825 830 820 825 830
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser GlyGly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly
835 840 845 835 840 845
Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys GlnGly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln
850 855 860 850 855 860
Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu GluLeu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu
865 870 875 880865 870 875 880
Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala TyrVal Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr
885 890 895 885 890 895
Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala ProAsp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro
900 905 910 900 905 910
Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu AsnGlu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn
915 920 925 915 920 925
Lys Ile Lys Met LeuLys Ile Lys Met Leu
930 930
<210> 9<210> 9
<211> 943<211> 943
<212> PRT<212> PRT
<213> Artificial Sequence<213> Artificial Sequence
<400> 9<400> 9
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly AlaMet Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 151 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu LeuLeu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30 20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala AlaLeu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45 35 40 45
Gln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala AspGln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala Asp
50 55 60 50 55 60
Tyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu GlyTyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu Gly
65 70 75 8065 70 75 80
Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser ThrTyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr
85 90 95 85 90 95
Val Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His AlaVal Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala
100 105 110 100 105 110
His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val AlaHis Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala
115 120 125 115 120 125
Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His GluVal Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu
130 135 140 130 135 140
Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu GluAla Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu
145 150 155 160145 150 155 160
Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln LeuAla Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu
165 170 175 165 170 175
Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr AlaAsp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala
180 185 190 180 185 190
Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro LeuVal Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
195 200 205 195 200 205
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyAsn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
210 215 220 210 215 220
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
225 230 235 240225 230 235 240
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn GlyAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
245 250 255 245 250 255
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
260 265 270 260 265 270
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
275 280 285 275 280 285
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProAsn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
290 295 300 290 295 300
Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
305 310 315 320305 310 315 320
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
325 330 335 325 330 335
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
340 345 350 340 345 350
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
355 360 365 355 360 365
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln
370 375 380 370 375 380
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
385 390 395 400385 390 395 400
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
405 410 415 405 410 415
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala LeuGlu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
420 425 430 420 425 430
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
435 440 445 435 440 445
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys GlnThr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
450 455 460 450 455 460
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
465 470 475 480465 470 475 480
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly GlyGly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
485 490 495 485 490 495
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
500 505 510 500 505 510
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn IleAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
515 520 525 515 520 525
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
530 535 540 530 535 540
Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser
545 550 555 560545 550 555 560
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProHis Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
565 570 575 565 570 575
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
580 585 590 580 585 590
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
595 600 605 595 600 605
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
610 615 620 610 615 620
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
625 630 635 640625 630 635 640
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
645 650 655 645 650 655
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
660 665 670 660 665 670
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
675 680 685 675 680 685
Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala LeuAsp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu
690 695 700 690 695 700
Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala AlaGlu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala
705 710 715 720705 710 715 720
Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg ProLeu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg Pro
725 730 735 725 730 735
Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Gly Ser Tyr AlaAla Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Gly Ser Tyr Ala
740 745 750 740 745 750
Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn GlyLeu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly
755 760 765 755 760 765
Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu GluGln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu
770 775 780 770 775 780
Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr AlaSer Lys Val Phe Ser Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala
785 790 795 800785 790 795 800
Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp AsnAsn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp Asn
805 810 815 805 810 815
Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr CysGly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys
820 825 830 820 825 830
Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala LysGly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys
835 840 845 835 840 845
Met Thr Val Val Pro Pro Glu Gly Ser Gly Gly Ser Thr Asn Leu SerMet Thr Val Val Pro Pro Glu Gly Ser Gly Gly Ser Thr Asn Leu Ser
850 855 860 850 855 860
Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln Glu SerAsp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser
865 870 875 880865 870 875 880
Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys ProIle Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro
885 890 895 885 890 895
Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp GluGlu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu
900 905 910 900 905 910
Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp AlaAsn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala
915 920 925 915 920 925
Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met LeuLeu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu
930 935 940 930 935 940
<210> 10<210> 10
<211> 997<211> 997
<212> PRT<212> PRT
<213> Artificial Sequence<213> Artificial Sequence
<400> 10<400> 10
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly AlaMet Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 151 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu LeuLeu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30 20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala AlaLeu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45 35 40 45
Gln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala AspGln Thr Ser Glu Ser Gly Gly Gly Gly Ser Pro Gly Ala Ala Ala Asp
50 55 60 50 55 60
Tyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu GlyTyr Lys Asp Asp Asp Asp Lys Gly Ser Val Asp Leu Arg Thr Leu Gly
65 70 75 8065 70 75 80
Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser ThrTyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr
85 90 95 85 90 95
Val Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His AlaVal Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala
100 105 110 100 105 110
His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val AlaHis Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala
115 120 125 115 120 125
Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His GluVal Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu
130 135 140 130 135 140
Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu GluAla Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu
145 150 155 160145 150 155 160
Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln LeuAla Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu
165 170 175 165 170 175
Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr AlaAsp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala
180 185 190 180 185 190
Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro LeuVal Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
195 200 205 195 200 205
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly GlyAsn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
210 215 220 210 215 220
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
225 230 235 240225 230 235 240
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn AsnAla His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
245 250 255 245 250 255
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
260 265 270 260 265 270
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
275 280 285 275 280 285
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProAsn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
290 295 300 290 295 300
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
305 310 315 320305 310 315 320
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
325 330 335 325 330 335
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
340 345 350 340 345 350
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
355 360 365 355 360 365
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
370 375 380 370 375 380
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
385 390 395 400385 390 395 400
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
405 410 415 405 410 415
Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala LeuAsp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
420 425 430 420 425 430
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
435 440 445 435 440 445
Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys GlnThr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
450 455 460 450 455 460
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
465 470 475 480465 470 475 480
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyGly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
485 490 495 485 490 495
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys GlnLys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
500 505 510 500 505 510
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn IleAla His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
515 520 525 515 520 525
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val LeuGly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
530 535 540 530 535 540
Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala SerCys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser
545 550 555 560545 550 555 560
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu ProHis Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
565 570 575 565 570 575
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala IleVal Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
580 585 590 580 585 590
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg LeuAla Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
595 600 605 595 600 605
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val ValLeu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
610 615 620 610 615 620
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val GlnAla Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
625 630 635 640625 630 635 640
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp GlnArg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
645 650 655 645 650 655
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu ThrVal Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
660 665 670 660 665 670
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr ProVal Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
675 680 685 675 680 685
Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala LeuAla Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
690 695 700 690 695 700
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly LeuGlu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
705 710 715 720705 710 715 720
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys GlnThr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
725 730 735 725 730 735
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala HisAla Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
740 745 750 740 745 750
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly GlyGly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
755 760 765 755 760 765
Arg Pro Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp ProArg Pro Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
770 775 780 770 775 780
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys LeuAla Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
785 790 795 800785 790 795 800
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly SerGly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
805 810 815 805 810 815
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly GlnPro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
820 825 830 820 825 830
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val PheSer Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
835 840 845 835 840 845
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr GluHis Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
850 855 860 850 855 860
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu GlyThr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
865 870 875 880865 870 875 880
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe ThrAla Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
885 890 895 885 890 895
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser GlyGly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly
900 905 910 900 905 910
Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys GlnGly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln
915 920 925 915 920 925
Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu GluLeu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu
930 935 940 930 935 940
Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala TyrVal Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr
945 950 955 960945 950 955 960
Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala ProAsp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro
965 970 975 965 970 975
Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu AsnGlu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn
980 985 990 980 985 990
Lys Ile Lys Met LeuLys Ile Lys Met Leu
995 995
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