CN114015697A

CN114015697A - 与遗传性心律失常有关的突变基因及其应用

Info

Publication number: CN114015697A
Application number: CN202111382083.1A
Authority: CN
Inventors: 刘哲; 梁庆渊; 赵娜娜; 赖开生; 刘昕超; 高璇; 李方玉; 侯青; 惠汝太
Original assignee: Bosinor Beijing Medical Laboratory Co ltd
Current assignee: Bosinor Beijing Medical Laboratory Co ltd
Priority date: 2021-11-22
Filing date: 2021-11-22
Publication date: 2022-02-08
Anticipated expiration: 2041-11-22
Also published as: CN114015697B

Abstract

本发明涉及人类遗传学和内科心血管技术领域，具体涉及了与遗传性心律失常有关的突变基因，与野生型RYR2基因编码DNA的参考序列相比，突变基因RYR2的核苷酸序列为SEQ ID NO:3；在基因组位置chr1:237608771处，碱基G突变为碱基T，在基因组位置chr1:237608774处，碱基C突变为碱基T；参考基因组版本是GRCh37。本发明还涉及上述突变基因在制备检测试剂盒中的应用。本发明提供的突变基因可以作为临床辅助诊断的生物标志物；检测该变异的携带者，为受试者提供优生优育指导和遗传咨询，减少患儿出生，对儿茶酚胺敏感性室性心动过速的早期诊断，或者辅助临床判断具有重要意义。

Description

与遗传性心律失常有关的突变基因及其应用

技术领域

本发明涉及人类遗传学和内科心血管技术领域，尤其涉及与遗传性心律失常有关的突变基因及其应用。

背景技术

儿茶酚胺敏感性室性心动过速(catecholaminergic polymorphic ventriculartachycardia,CPVT)是一种遗传性心律失常疾病，发病年龄平均为8岁，是心脏结构正常而对儿茶酚胺敏感的遗传性疾病。主要临床表现为：体力活动、压力或儿茶酚胺注射诱发的多形性室性心动过速，晕厥，猝死，窦房结功能障碍，房室结功能障碍，心房颤动，心房停搏，左室功能不全，左室扩张，癫痫。

目前已发现2个与CPVT有关的致病基因，分别为常染色体显性遗传，编码心脏兰尼碱受体2型的RYR2基因(称为CPVT1型)和常染色体隐性遗传，编码心脏肌贮钙蛋白的CASQ2基因(称为CPVT2型)。在已知2个CPVT致病基因中，约65％先证者存在RYR2突变，3-5％先证者为CASQ2突变。65％诊断为CPVT患者基因筛查为阳性。

CPVT专家共识建议:CPVT1(RYR2)和CPVT2(CASQ2)的基因检测推荐：基于病史，家族史，以及运动或儿茶酚胺应激诱发的ECG阳性表型，具有CPVT临床证据的患者，都推荐进行基因检测。

RYR2基因编码心肌细胞肌浆网Ca²⁺释放通道-雷诺定受体2(Ryanodine receptor2，RyR2)，该基因的突变引起RYR2通道舒张期肌浆网Ca²⁺泄露增加，引发心肌细胞Ca²⁺稳态失衡，从而导致致死性心律失常。

目前仍存在大量未知的RYR2基因突变位点，进一步发现新的RYR2基因的突变位点对于研究儿茶酚胺敏感性室性心动过速的发病机制，对儿茶酚胺敏感性室性心动过速的早期诊断，或者辅助临床判断具有重要意义。

发明内容

本发明的目的是针对儿茶酚胺敏感性室性心动过速，提供一种与遗传性心律失常有关的突变基因及其应用。

本发明提供的技术方案如下：

一、本发明提供与遗传性心律失常有关的突变基因，与野生型RYR2基因编码DNA的参考序列相比，突变基因RYR2的核苷酸序列为SEQ ID NO:3；在基因组位置chr1:237608771处，碱基G突变为碱基T，在基因组位置chr1:237608774处，碱基C突变为碱基T；参考基因组版本是GRCh37。

二、本发明还提供了上述与遗传性心律失常有关的突变基因在制备检测试剂盒中的应用。

优选地，所述检测试剂盒包括引物SEQ ID NO:1和SEQ ID NO:2。

优选地，所述检测试剂盒还包括Taq DNA聚合酶和PCR缓冲液。

三、本发明的原理和有益效果在于：

本发明公开的与遗传性心律失常有关的突变基因可以作为临床辅助诊断儿茶酚胺敏感性室性心动过速的生物标志物，对儿茶酚胺敏感性室性心动过速的早期诊断，或者辅助临床判断具有重要意义；基于与遗传性心律失常有关的突变基因开发的检测试剂盒，可以检测出携带突变基因的患者，为受试者提供优生优育指导和遗传咨询，减少患儿出生。

附图说明

图1为实施例3的家系图；

图2为实施例3中携带c.1241G>T(RYR2:p.Arg414Leu het)杂合错义的先证者的Sanger测序图；

图3为实施例3家系中未携带c.1241G>T(RYR2:p.Arg414Leu het)杂合错义的先证者父亲等人的Sanger测序图；

图4为实施例3中携带c.1244C>T(RYR2:p.Thr415Ile het)杂合错义的先证者的Sanger测序图；

图5为实施例3家系中未携带c.1244C>T(RYR2:p.Thr415Ile het)杂合错义的先证者母亲等人的Sanger测序图。

具体实施方式

下面通过具体实施方式进一步详细说明：

实施例1-与遗传性心律失常有关的突变基因

与遗传性心律失常有关的突变基因，具体突变如下表1所示：

表1与遗传性心律失常有关的突变基因的具体检测结果

(1)在基因组位置chr1:237608722-chr1:237608821处，野生型RYR2基因的序列为：

为野生型RYR2基因在基因组chr1:237608771处的碱基，

为野生型RYR2基因在基因组chr1:237608774处的碱基。

在基因组位置chr1:237608722-chr1:237608821处，突变基因RYR2的序列为：

为突变基因RYR2在基因组chr1:237608771处和基因组chr1:237608774处的碱基处的碱基。

(2)野生型RYR2基因编码DNA的参考序列为：

为野生型RYR2基因编码DNA参考序列的第1241位突变前碱基。

为野生型RYR2基因编码DNA参考序列的第1244位突变前碱基。

c.1241G>T表示：与野生型RYR2基因编码DNA的参考序列相比，突变基因RYR2的第1241位点的碱基G突变为碱基T。

c.1244C>T表示：与野生型RYR2基因编码DNA的参考序列相比，突变基因RYR2的第1244位的碱基C突变为碱基T，突变基因RYR2的具体核苷酸序列为SEQ ID NO:3。

(4)野生型RYR2基因编码蛋白为：

为野生型RYR2基因编码蛋白氨基酸序列的第414位精氨酸(Arg,R)，以及第415位苏氨酸(Thr,T)。

p.Arg414Leu表示：与野生型RYR2基因编码蛋白的氨基酸序列相比，突变基因RYR2编码蛋白的氨基酸第414位的精氨酸(Arg,R)突变为亮氨酸(Leu,L)。

p.Thr415Ile表示：与野生型RYR2基因编码蛋白的氨基酸序列相比，突变基因RYR2编码蛋白的氨基酸第415位的苏氨酸(Thr,T)突变为异亮氨酸(Ile,I)，突变基因RYR2编码蛋白的具体氨基酸序列为SEQ ID NO:4。

(5)查询人群频率数据库发现RYR2基因c.1241G>T杂合错义变异(RYR2:p.Arg414Leu het)为罕见变异(千人基因组：无，ESP6500：无，ExAC：无)。采用多个生物信息预测软件(包括SIFT和Polyphen-2等)交叉预测，结果多为有害(SIFT为“D”，Polyphen-2为“B”，MutationTaster_pred为“D”，VEST3评分为“0.598”，其他为“5个D/1个M”)，提示该变异所致的氨基酸改变可能会对蛋白功能产生影响。氨基酸由极性带正电荷的精氨酸变为非极性的亮氨酸。查询数据库发现该位置的氨基酸在脊椎动物中高度保守。查询ClinVar数据库发现该变异被上报者评定为“Uncertain_significance”，HGMD数据库发现该变异被评定为“DiseaseCa using mutation”，该位点附近的错义变异c.1250G>T(p.Arg417Leu)被上报者评定为疑似致病突变(相关疾病：not provided)，c.1258C>T(p.Arg420Trp)和c.1259G>A(p.Arg420Gln)被上报者评定为儿茶酚胺敏感性室性心动过速的致病突变(ClinVar数据库)。根据现有证据：该变异为罕见变异、软件预测该变异可能会对蛋白功能产生影响、该位置氨基酸在脊椎动物中高度保守、附近位点多次被报导为致病突变，但缺乏家系连锁及功能学证据支持，所以该变异为儿茶酚胺敏感性室性心动过速的高度可疑致病突变。

查询人群频率数据库发现RYR2基因c.1244C>T杂合错义变异(RYR2:p.Thr415Ilehet)为罕见变异(千人基因组：无，ESP6500：无，ExAC：无)。采用多个生物信息预测软件(包括SIFT和Polyphen-2等)交叉预测，结果多为有害(SIFT为“D”，Polyphen-2为“B”，MutationTaster_pred为“D”，VEST3评分为“0.524”，其他为“5个D/1个M”)，提示该变异所致的氨基酸改变可能会对蛋白功能产生影响。氨基酸由极性不带电荷的苏氨酸变为非极性的异亮氨酸。查询数据库发现该位置的氨基酸在脊椎动物中高度保守。查询ClinVar、HGMD数据库未发现该变异，该位点附近的错义变异c.1250G>T(p.Arg417Leu)被上报者评定为疑似致病突变(相关疾病：not provided)，c.1258C>T(p.Arg420Trp)和c.1259G>A(p.Arg420Gln)被上报者评定为儿茶酚胺敏感性室性心动过速的致病突变(ClinVar数据库)，文献检索未发现该变异与疾病相关的报导。根据现有证据：该变异为罕见变异、软件预测该变异可能会对蛋白功能产生影响、该位置氨基酸在脊椎动物中高度保守、附近位点多次被报导为致病突变，但缺乏家系连锁及功能学证据支持，所以可疑致病变异。

实施例2-与遗传性心律失常有关的突变基因的检测试剂盒

与遗传性心律失常有关的突变基因的检测试剂盒，包括Taq DNA聚合酶、PCR缓冲液和引物等。具体引物如下：

上游引物(RYR2-E14F，SEQ ID NO:1)：5'AATGAATAAATGGAATGGCTA 3'；

下游引物(RYR2-E14R，SEQ ID NO:2)：5'GTTTCTGCTGACTTACCAA 3'；

长度：451bp。

利用本试剂盒筛选突变的致病基因RYR2的具体步骤为：提取待测者DNA，然后使用经设计的引物组合(SEQ ID NO:1和SEQ ID NO:2)对RYR2基因进行扩增，得到PCR产物，使用1.5％的琼脂糖凝胶电泳检测PCR产物，选用1000bp Marker作为参考，检测验证扩增产物为预期的大小，最后对PCR产物进行测序。从NCBI(https://www.ncbi.nlm.nih.gov/)数据库获得参考序列和测序结果进行比对，判断待测者RYR2基因是否携带c.1241G>T、c.1244C>T杂合错义变异，协助临床确诊儿茶酚胺敏感性室性心动过速。

实施例3-家系验证实验

本实施例采用家系连锁分析方法验证与遗传性心律失常有关的突变基因的致病性。

具体地，选取一个家族性儿茶酚胺敏感性室性心动过速家系中的三代成员，该家系中的先证者(男，15岁)临床被诊断为儿茶酚胺敏感性室性心动过速。

在先证者及其家属自愿签署知情同意书的前提下，寄送5-10mL全血样本，建立病历资料库，详细记录先证者病情、家系情况等资料。本研究已得到本单位伦理委员会批准。

先证者临床概况描述：

表3先证者临床概况

采用实施例2提供的体外检测试剂盒对先证者及其家属的RYR2基因进行基因检测，结果如图1-图5所示，图1为实施例3的家系图；图2为实施例3中携带c.1241G>T(RYR2:p.Arg414Leu het)杂合错义的先证者的Sanger测序图；图3为实施例3家系中未携带c.1241G>T(RYR2:p.Arg414Leu het)杂合错义的先证者父亲等人的Sanger测序图；图4为实施例3中携带c.1244C>T(RYR2:p.Thr415Ile het)杂合错义的先证者的Sanger测序图；图5为实施例3家系中未携带c.1244C>T(RYR2:p.Thr415Ile het)杂合错义的先证者母亲等人的Sanger测序图。

如图1-图5所示，先证者携带了常染色体显性遗传的儿茶酚胺敏感性室性心动过速的可疑致病变异RYR2基因c.1241G>T杂合错义变异(RYR2:p.Arg414Leu het)和RYR2基因c.1244C>T杂合错义变异(RYR2:p.Thr415Ile het)，家系验证以上变异为新发变异，父母均未携带，先证者为以上变异的复合杂合子；以上变异具备导致先证者出现临床表型的条件。

实施例4-针对家系外正常人的验证实验

采用实施例2的儿茶酚胺敏感性室性心动过速检测试剂盒，对3500例家系外正常人进行RYR2基因检测，结果均未能检测到该突变。

实施例5-针对家系外家族遗传性儿茶酚胺敏感性室性心动过速患者的验证实验

在中国范围内，对患有儿茶酚胺敏感性室性心动过速、肥厚型心肌病、扩张型心肌病、长QT等遗传病的患者中检测RYR2基因，患者总共4000例，每种疾病的患病人数不等，结果显示，除实施例3提供的家系外，仅在临床诊断患有儿茶酚胺敏感性室性心动过速的4例患者中检测到携带突变RYR2基因的c.1241G>T杂合错义变异(RYR2:p.Arg414Leu het)或RYR2基因c.1244C>T杂合错义变异(RYR2:p.Thr415Ile het)。

本实验再次验证说明突变的RYR2致病基因的会导致儿茶酚胺敏感性室性心动过速，支持临床诊断。

以上详细描述了本发明的较佳具体实施例。应当理解，本领域的普通技术人员无需创造性劳动就可以根据本发明的构思作出诸多修改和变化。因此，凡本技术领域中技术人员依本发明的构思在现有技术的基础上通过逻辑分析、推理或者有限的实验可以得到的技术方案，皆应在由权利要求书所确定的保护范围内。

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ttagacaaac atggaagaaa tcacaaggtt ctggatgtct tgtgctcact ctgtgtttgc 1860

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gatctcttct cctatggatt tgatggcctt catctctggt caggttgtat tgctcgtact 2220

gtaagctcac caaaccaaca tctgttaaga actgatgatg tcatcagttg ctgtttagat 2280

ctgagtgccc caagcatctc gttccgaatt aatggacaac ctgttcaagg aatgtttgag 2340

aatttcaaca tcgatggcct cttctttcca gtcgttagtt tctctgcagg aataaaagta 2400

cgctttctgc ttggagggcg acatggagaa ttcaaatttc ttcctccacc tgggtatgct 2460

ccttgttatg aagctgttct gccaaaagaa aagttgaaag tggaacacag ccgagagtac 2520

aagcaagaaa gaacttacac acgcgacctg ctgggcccca cagtttccct gacgcaagct 2580

gccttcacac ccatccctgt ggataccagc cagatcgtgt tgcctcctca tctagaaaga 2640

ataagagaaa aactggcaga gaatatccat gaactctggg ttatgaataa aattgagctt 2700

ggctggcagt atggtccggt tagagatgac aacaagagac aacacccatg cctggtggag 2760

ttctccaagc tgcctgaaca ggagcgcaat tacaacttac aaatgtcgct tgagaccctg 2820

aagactttgt tggcattagg atgtcatgtg ggtatatcag atgaacatgc tgaagacaag 2880

gtgaaaaaaa tgaagctacc caagaattac cagctgacaa gtggatacaa gcctgcccct 2940

atggacctga gctttatcaa actcacccca tcacaagaag caatggtgga caagttggca 3000

gaaaatgcac ataatgtgtg ggcgcgggat cgaatccggc agggctggac ttatggcatc 3060

caacaggacg taaagaacag aagaaatcct cgccttgttc cctacactct tctggatgac 3120

cgaaccaaga aatccaacaa ggacagcctc cgcgaggctg tgcgcacgct gctggggtac 3180

ggctacaact tggaagcacc agatcaagat catgcagcca gagccgaagt gtgcagcggc 3240

accggggaaa ggttccgaat cttccgtgcc gagaagacct atgcagtgaa ggccggacgg 3300

tggtattttg aatttgagac ggtcactgct ggagacatga gggttggttg gagtcgtcct 3360

ggttgtcaac cggatcagga gcttggctca gatgaacgtg cctttgcctt tgatggcttc 3420

aaggcccagc ggtggcatca gggcaatgaa cactatgggc gctcttggca agcaggcgat 3480

gtcgtggggt gtatggttga catgaacgaa cacaccatga tgttcacact gaatggtgaa 3540

atccttcttg atgattcagg ctcagaactg gctttcaagg actttgatgt tggcgatgga 3600

ttcatacctg tgtgtagcct tggagtggct caagtgggta ggatgaactt tggaaaggat 3660

gtcagcacct tgaaatattt caccatctgt ggcttacaag agggctatga accatttgcc 3720

gttaatacaa acagggatat taccatgtgg ctgagcaaga ggcttcctca gtttcttcaa 3780

gttccatcaa accatgaaca tatagaggtg accagaatag acggcaccat agacagttcc 3840

ccatgtttaa aggtcactca gaagtctttt ggttctcaga acagcaacac tgatatcatg 3900

ttttatcgcc tgagcatgcc gatcgagtgc gcggaggtct tctccaagac ggtggctgga 3960

gggctccctg gggctggcct ttttgggccc aagaatgact tggaagatta tgatgctgat 4020

tctgactttg aggttctgat gaagacagct catggccatc tagtgcccga tcgtgttgac 4080

aaagacaaag aagctactaa accagagttt aacaaccaca aagattatgc ccaggaaaag 4140

ccctctcgtc tgaaacaaag atttttgctt agaagaacaa agccagatta cagcacaagc 4200

cattctgcaa gactcaccga agatgtcctt gctgatgatc gggatgacta tgatttcttg 4260

atgcaaacgt ccacgtacta ttactcagtg agaatctttc ctggacaaga acctgctaat 4320

gtctgggtgg gctggattac atcagatttc catcagtatg acacaggctt tgacttggac 4380

agagttcgca cagtaacagt tactctagga gatgaaaaag gaaaagtgca tgaaagcatc 4440

aaacgcagca actgctatat ggtatgtgcg ggtgagagca tgagccccgg gcaaggacgc 4500

aacaataatg gactggagat tggctgtgtg gtggatgctg ccagcgggct gctcacattc 4560

attgccaatg gcaaggaact gagcacatac tatcaggtgg aaccgagtac aaaattattt 4620

cctgcggttt ttgcacaagc tacaagtccc aatgttttcc agtttgagtt gggaagaata 4680

aagaatgtga tgcctctctc ggcgggatta ttcaagagtg agcacaagaa ccccgtgccg 4740

cagtgccccc cgcgcctcca cgtgcagttc ctgtcacacg tcctgtggag cagaatgccc 4800

aaccagtttt tgaaggtaga tgtgtctcga ataagtgaac gccaaggctg gttggtgcag 4860

tgtttggatc ctctgcagtt catgtctctt catatccctg aggaaaacag atctgttgac 4920

atcttagagt tgacagagca ggaggaattg ctgaaatttc actatcacac tctccggctc 4980

tactcagccg tctgtgctct tgggaaccac cgggtggccc atgccctgtg cagccatgtg 5040

gatgaacctc agctcctcta tgccattgag aacaagtaca tgcctggttt gctgcgtgct 5100

ggctactatg acctgctgat tgacatccac ctgagctcct atgccactgc caggctcatg 5160

atgaacaacg agtacattgt ccccatgacg gaggagacga agagcatcac cctgttccct 5220

gatgagaaca aaaaacacgg ccttccaggg atcggcctca gcacctccct caggccacgg 5280

atgcagtttt cctcccccag ttttgtaagc attagtaatg aatgttacca gtacagtcca 5340

gagttcccac tggacatcct caagtccaaa accatacaga tgctgacaga agctgttaaa 5400

gagggcagtc ttcatgcccg ggacccagtt ggagggacta ctgaattcct ctttgtacct 5460

ctcatcaagc ttttctatac cctgctgatc atgggcatct ttcacaacga ggacttgaag 5520

cacatcttgc agttgattga gcccagtgtg tttaaagaag ctgccactcc ggaggaggag 5580

agtgacacgc tggagaaaga gctcagtgtg gacgatgcaa agctgcaagg agctggtgag 5640

gaagaagcca aggggggcaa gcggcccaag gaaggcctgc tccaaatgaa actgccagag 5700

ccagttaaat tgcagatgtg cctactgctt cagtacctct gtgactgcca ggtccggcac 5760

cggatagaag ccattgtagc cttttcagat gattttgtgg ctaagctcca agacaatcaa 5820

cgtttccgat acaacgaagt catgcaagcc ttaaacatgt cagctgcact cacagccagg 5880

aagacaaagg aatttagatc accacctcaa gaacagatca atatgcttct caattttaag 5940

gatgacaaaa gtgaatgtcc atgtccagaa gaaattcgtg accaactatt ggatttccat 6000

gaagatttga tgacacattg tggaattgag ctggatgaag atgggtctct ggatggaaac 6060

agtgatttaa caattagagg gcgtctgcta tccctggtag aaaaggtgac atatctgaag 6120

aagaagcaag cagaaaaacc agttgagagt gactccaaaa agtcctccac tctgcagcag 6180

ctgatttctg agaccatggt ccgatgggct caggagtctg tcattgaaga ccccgagctg 6240

gtgagggcca tgtttgtgtt gctccatcgg cagtatgacg gcattggggg tcttgttcgg 6300

gccctgccaa agacctacac gataaatggt gtgtccgtgg aggacaccat caacctgctg 6360

gcatcccttg gtcagattcg gtccctgctg agtgtgagaa tgggcaaaga agaagagaag 6420

ctcatgattc gtggattagg ggatattatg aataacaaag tgttttacca gcaccctaat 6480

ctcatgaggg cactggggat gcacgagact gtgatggagg tcatggtgaa cgtccttgga 6540

ggtggagagt ccaaggaaat cacctttccc aagatggtgg ccaactgttg ccgttttctc 6600

tgttacttct gtcgtataag taggcagaat caaaaagcta tgtttgatca tctcagttat 6660

ttactggaaa acagcagtgt tggtcttgcc tccccagcta tgagaggttc aacaccactg 6720

gatgtggctg cagcttcggt gatggataat aatgaactag cattagctct gcgtgagccg 6780

gatctagaaa aggtagttcg ttatttggct ggttgtggac tgcaaagttg ccagatgctg 6840

gtgtctaagg gctatccaga cattgggtgg aacccagttg aaggagagag atatcttgac 6900

tttcttagat ttgctgtctt ctgtaatggg gagagtgtgg aggaaaatgc aaatgtcgtg 6960

gtgagattgc tcattcggag gcctgagtgt tttggtcctg ctttgagagg agaaggtggg 7020

aatgggcttc ttgcagcaat ggaagaagcc atcaaaatcg ccgaggatcc ttcccgagat 7080

ggtccctcac caaatagcgg atccagtaaa acacttgaca cagaggagga ggaagatgac 7140

actatccaca tggggaacgc gatcatgacc ttctattcag ctttgattga cctcttggga 7200

cgctgtgctc ctgagatgca tttgattcat gccgggaagg gagaagccat cagaattagg 7260

tccattttga gatccctcat tcccctggga gatttggtgg gcgttatcag catcgctttt 7320

cagatgccaa caatagccaa agatgggaat gtggtggaac ctgacatgtc tgcggggttt 7380

tgcccagatc acaaggcagc catggtttta ttccttgaca gggtctatgg gattgaggtt 7440

caagacttcc tcctccatct tcttgaggtt ggctttctgc cagatctccg ggcggctgct 7500

tctttagata cggcagcttt gagtgctaca gacatggcct tggccctcaa tcggtacctt 7560

tgcacagccg tcttgccatt gttaacaaga tgtgctcctc tctttgctgg cacagagcac 7620

cacgcttctc tcattgactc attacttcat actgtgtata gactttctaa gggctgttca 7680

cttaccaaag ctcagcggga ttccatagaa gtttgtttac tctctatttg tggacaactg 7740

agaccttcta tgatgcagca cttactcaga agattagtat ttgatgttcc attattaaat 7800

gaacacgcaa agatgcctct taaactgctg acaaatcatt atgaaagatg ctggaaatat 7860

tactgcctgc ctggagggtg gggaaacttt ggtgctgcct cagaagaaga acttcattta 7920

tcaagaaagt tgttctgggg catttttgat gccctgtctc aaaagaaata tgaacaagaa 7980

cttttcaaac tggcactgcc ttgcctgagt gcagttgcgg gagctttgcc tccagactac 8040

atggagtcaa attatgtcag tatgatggaa aaacagtcat caatggattc tgaagggaac 8100

tttaacccac aacctgttga tacctcaaat attacaattc ctgagaaatt ggaatacttc 8160

attaacaaat atgcagaaca ctcccatgac aaatggtcaa tggacaagtt ggcaaatgga 8220

tggatttatg gagaaatata ttcagactct tctaaggttc agccattaat gaagccatat 8280

aagctattgt ctgaaaagga aaaagaaatt tatcgctggc caatcaaaga atctttaaaa 8340

actatgctgg cttggggctg gagaattgaa agaactcggg agggagacag catggccctt 8400

tacaaccgga ctcgtcgtat ttctcagaca agccaggttt ctgtggacgc tgcccatggt 8460

tacagtcccc gggccattga catgagcaat gttacactat ctagagacct gcatgctatg 8520

gcagaaatga tggctgaaaa ctaccataat atatgggcaa agaaaaagaa aatggagttg 8580

gagtccaaag gaggaggaaa ccatcctctg ctggtgccct atgatacact gacagccaaa 8640

gagaaagcca aggatagaga aaaagcacag gacatcctca agttcttgca gatcaatgga 8700

tatgctgtat ccagaggatt taaggacctg gaactggaca cgccttctat tgagaaacga 8760

tttgcctata gtttcctcca acaactcatt cgctatgtgg atgaagccca tcagtatatc 8820

ctggagtttg atggtggcag cagaggcaaa ggagaacatt tcccttatga acaagaaatc 8880

aagttctttg caaaagtcgt tcttccttta attgatcagt atttcaaaaa ccatcgttta 8940

tacttcttat ctgcagcaag cagacctctc tgctctggag gacatgcttc caacaaagag 9000

aaagaaatgg tgactagcct attctgcaaa cttggagttc ttgtcaggca taggatttca 9060

ctatttggca atgatgcaac atcaattgtc aactgtcttc atattttggg tcagactttg 9120

gatgcaagga cagtgatgaa gactggcctg gagagtgtta aaagtgcact cagagctttt 9180

ctggacaacg ctgcagagga tctggagaag accatggaaa acctcaagca gggccagttc 9240

actcacaccc gaaaccagcc caaaggggtt actcagatta tcaattacac cacagtggcc 9300

ctgctgccaa tgctgtcttc attatttgaa catattggcc agcatcagtt cggagaagac 9360

ctaatattgg aagatgtcca ggtgtcttgt tatagaattc tgactagctt atatgctttg 9420

ggaaccagca agagtattta cgtggagagg caacgttctg cattaggaga atgtctagct 9480

gcctttgctg gtgcttttcc tgtagcattt ttggaaactc atctggacaa acataatatt 9540

tactccatct acaataccaa gtcttcacga gaaagagcag ctctcagttt gccaactaat 9600

gtggaagatg tttgtccaaa cataccgtct ttggagaaac tcatggaaga aatcgtggaa 9660

ttagccgagt ccggcattcg ctacactcaa atgccacatg tcatggaagt catactgccc 9720

atgctttgca gctacatgtc tcgttggtgg gagcatggac ctgagaacaa tccagaacgg 9780

gccgagatgt gctgcacagc cctgaactca gagcacatga acacacttct agggaacata 9840

ttgaaaatca tatataataa cttggggatt gatgagggag cctggatgaa gaggctagca 9900

gtgttttccc agcctataat aaataaagtg aaacctcagc tcttgaaaac tcatttcttg 9960

ccgttaatgg agaaactcaa gaaaaaggca gctacggtgg tgtctgagga agaccacctg 10020

aaagctgagg ccagggggga catgtcggag gcagaactcc tcatcctaga tgagttcacc 10080

acactggcca gagatctcta tgccttctac cctctcttga ttagatttgt ggactataac 10140

agggcaaagt ggctaaagga gcctaaccca gaagcagagg agctcttccg catggtggct 10200

gaagtgttta tctactggtc gaagtcccat aatttcaaaa gagaagagca gaacttcgtt 10260

gtacagaatg aaatcaacaa tatgtctttc cttattactg ataccaagtc aaagatgtca 10320

aaggcagctg tttctgatca ggaaaggaag aaaatgaagc gcaaaggaga tcggtattcc 10380

atgcagacct ctctgattgt agcagctctg aagcggttac tgcccattgg gttgaacatc 10440

tgtgcccctg gggaccagga gctcattgct ctggccaaaa atcgatttag cctgaaagat 10500

accgaggatg aagtacgaga tataatccgc agcaatattc atttacaagg caagttggag 10560

gatcctgcta ttagatggca aatggctctt tacaaagact taccaaacag gactgatgat 10620

acctcagatc cagagaagac ggtagaaaga gtattggata tagcaaatgt gctttttcat 10680

cttgaacaga agtctaaacg tgtgggtcgg agacattact gtctggtgga acatcctcag 10740

agatctaaaa aggctgtatg gcataaacta ctgtccaagc agaggaaaag ggctgttgta 10800

gcctgcttcc ggatggcccc cttatataat ctgccaaggc atcgggctgt caatctcttt 10860

cttcagggat atgaaaagtc ttggattgaa acagaagaac attactttga agataaactg 10920

atagaagatt tagcaaaacc tggggctgaa cctccagaag aagatgaagg cactaagaga 10980

gttgatcctc tacatcagct gatccttctg tttagtcgga cagctttaac agagaaatgc 11040

aaactggagg aagatttttt atatatggcc tatgcagata ttatggcaaa gagttgtcat 11100

gatgaggaag atgacgatgg tgaagaggaa gtgaagagtt ttgaagaaaa agaaatggaa 11160

aagcaaaagc ttctatacca gcaagcccga ctccacgatc gtggcgcggc tgagatggtg 11220

ctacagacaa tcagtgccag caaaggtgaa actggaccaa tggtagcagc tactctgaaa 11280

cttggaattg ctattttaaa tggtgggaac tccacagtac agcagaaaat gcttgactac 11340

ctcaaggaga aaaaggatgt gggcttcttt cagagcctgg ccggcctgat gcagtcatgt 11400

agtgtccttg acctaaatgc atttgagcga caaaacaaag ctgaaggtct tgggatggtg 11460

acagaggaag gatcaggaga aaaggttctg caggacgatg agttcacctg tgacctcttc 11520

cgattcctgc aactactctg tgagggacac aactcagatt ttcagaatta tctgagaact 11580

cagactggca ataatacaac tgtcaacata attatctcca ctgtagacta cctactgaga 11640

gttcaggaat caattagtga cttttattgg tattactctg ggaaagatgt tattgatgaa 11700

caaggacaac ggaatttctc caaagctatc caagtggcaa aacaagtctt taacactctt 11760

acagagtata ttcagggtcc ttgcactggg aatcaacaga gtttggcaca cagcaggctg 11820

tgggatgctg tggtcggctt tcttcatgtg tttgcccata tgcagatgaa gctgtcgcag 11880

gattccagtc aaattgagct attaaaagaa ttaatggatc tgcagaagga tatggtggtc 11940

atgttgctgt ccatgttaga aggtaatgtt gttaatggaa cgattggcaa acagatggtg 12000

gatatgcttg tggaatcttc caacaacgtg gagatgattc tcaaattttt tgacatgttc 12060

ttaaaactaa aggatttgac gtcgtctgat acttttaaag aatatgaccc cgatggcaag 12120

ggagtcattt ccaagaggga cttccacaaa gcgatggaga gccataagca ctacacgcag 12180

tcagaaacgg aatttctttt gtcttgtgcg gagacggatg agaatgaaac cctcgactac 12240

gaagagttcg tcaaacgctt ccacgaacct gcgaaggaca tcggcttcaa cgtcgccgtc 12300

cttctgacaa acctctctga gcacatgccc aacgataccc gacttcagac ttttctggaa 12360

ttagcagaga gcgtcctgaa ttatttccag ccctttctgg gccgcatcga aatcatggga 12420

agcgccaaac gcatcgagag ggtctatttt gaaatcagtg agtccagccg aacccagtgg 12480

gagaagcccc aggtcaagga gtccaaaaga cagttcatat ttgacgtggt caacgaaggc 12540

ggagagaaag agaagatgga actctttgtg aacttctgcg aggacaccat ctttgaaatg 12600

cagctggcgg ctcagatctc ggagtcggac ttgaacgaga ggtcagcgaa taaggaagaa 12660

agcgagaagg agaggccgga agagcagggg ccgaggatgg ctttcttctc cattctgacg 12720

gtcaggtcgg ccctgtttgc gctcaggtac aatatcttga cccttatgcg aatgctcagt 12780

ctgaagagcc tgaagaagca gatgaaaaaa gtaaaaaaga tgaccgtgaa ggacatggtc 12840

acggccttct tttcatccta ctggagtatt ttcatgaccc tcttgcactt cgtggccagc 12900

gttttcagag gctttttccg catcatttgc agcctgctgc ttgggggaag cctcgtcgaa 12960

ggtgctaaaa agatcaaagt tgcagaactg ttagccaaca tgccagaccc cactcaggat 13020

gaggttagag gagatgggga ggagggagag aggaaacccc tggaagccgc cctgccctcc 13080

gaggatctga ccgacttaaa ggagctgaca gaggaaagtg accttctttc ggacatcttt 13140

ggcctggatc tgaagagaga aggaggacag tacaaactga ttcctcataa tccaaatgct 13200

gggctcagtg acctcatgag caacccagtc cccatgcctg aggtgcagga aaaatttcag 13260

gaacagaagg caaaagaaga agaaaaggaa gaaaaagaag aaaccaaatc tgaacctgaa 13320

aaagccgagg gagaagatgg agaaaaagaa gagaaagcca aggaagacaa gggcaaacaa 13380

aagttgaggc agcttcacac acacagatac ggagaaccag aagtgccaga gtcagcattc 13440

tggaagaaaa tcatagcata tcaacagaaa cttctaaact attttgctcg caacttttac 13500

aacatgagaa tgttagcctt atttgtcgca tttgctatca atttcatctt gctcttttat 13560

aaggtctcca cttcttctgt ggttgaagga aaggagctcc ccacgagaag ttcaagtgaa 13620

aatgccaaag tgacaagcct ggacagcagc tcccatagaa tcatcgcagt tcactatgta 13680

ctagaggaga gcagcggcta catggagccc acgttgcgta tcttagctat tctgcacacg 13740

gtcatttctt tcttctgcat cattggatac tactgcttga aagtcccatt ggttattttt 13800

aagcgagaaa aggaagtggc acggaaattg gaatttgatg ggctttatat tacagaacag 13860

ccttcagaag atgatattaa aggccagtgg gatagactcg taatcaacac acagtcattt 13920

cccaacaact actgggacaa atttgttaaa agaaaggtta tggataaata tggagagttc 13980

tacggccgag acagaatcag tgaattactt ggcatggaca aggcagctct ggacttcagt 14040

gatgccagag aaaagaagaa gccaaagaaa gacagctcct tatcagctgt actgaactcc 14100

attgatgtga agtatcagat gtggaaacta ggagtcgttt tcactgacaa ctccttcctc 14160

tacctagcct ggtatatgac tatgtctgtt cttggacact ataacaactt tttttttgcc 14220

gctcaccttc tcgacattgc tatgggattc aagacattaa gaaccatctt gtcctcagta 14280

actcacaatg gcaaacagct cgtattaacc gttggcttat tagctgttgt tgtataccta 14340

tacactgtgg tggcattcaa ttttttccga aaattctaca ataaaagtga agatggtgat 14400

acaccagata tgaaatgtga cgatatgcta acatgctata tgttccacat gtatgttgga 14460

gttcgtgctg gaggagggat cggggatgaa atcgaagacc cagcaggaga tgaatatgag 14520

atctatcgaa tcatctttga catcactttc ttcttctttg ttattgtcat tctcttggcc 14580

ataatacaag gtctaattat tgatgctttt ggagaactaa gagaccaaca ggaacaagtc 14640

aaagaagaca tggagaccaa atgcttcatc tgtgggatag gcaatgatta cttcgacaca 14700

gtgccacatg gctttgaaac ccacacttta caggagcaca acttggctaa ttacttgttt 14760

tttctgatgt atcttataaa caaagatgaa acagaacaca caggacagga atcttatgtc 14820

tggaagatgt atcaagaaag gtgttgggaa tttttcccag caggggattg cttccggaaa 14880

cagtatgaag accagctaaa ttaa 14904

<210> 4

<211> 4967

<212> PRT

<213> homo sapiens

<400> 4

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

1 5 10 15

Asp Asp Glu Val Val Leu Gln Cys Thr Ala Thr Ile His Lys Glu Gln

20 25 30

Gln Lys Leu Cys Leu Ala Ala Glu Gly Phe Gly Asn Arg Leu Cys Phe

35 40 45

Leu Glu Ser Thr Ser Asn Ser Lys Asn Val Pro Pro Asp Leu Ser Ile

50 55 60

Cys Thr Phe Val Leu Glu Gln Ser Leu Ser Val Arg Ala Leu Gln Glu

65 70 75 80

Met Leu Ala Asn Thr Val Glu Lys Ser Glu Gly Gln Val Asp Val Glu

85 90 95

Lys Trp Lys Phe Met Met Lys Thr Ala Gln Gly Gly Gly His Arg Thr

100 105 110

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

115 120 125

Tyr Leu Cys Cys Leu Ser Thr Ser Arg Ser Ser Thr Asp Lys Leu Ala

130 135 140

Phe Asp Val Gly Leu Gln Glu Asp Thr Thr Gly Glu Ala Cys Trp Trp

145 150 155 160

Thr Ile His Pro Ala Ser Lys Gln Arg Ser Glu Gly Glu Lys Val Arg

165 170 175

Val Gly Asp Asp Leu Ile Leu Val Ser Val Ser Ser Glu Arg Tyr Leu

180 185 190

His Leu Ser Tyr Gly Asn Gly Ser Leu His Val Asp Ala Ala Phe Gln

195 200 205

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

210 215 220

Gln Gly Tyr Leu Ile Gly Gly Asp Val Leu Arg Leu Leu His Gly His

225 230 235 240

Met Asp Glu Cys Leu Thr Val Pro Ser Gly Glu His Gly Glu Glu Gln

245 250 255

Arg Arg Thr Val His Tyr Glu Gly Gly Ala Val Ser Val His Ala Arg

260 265 270

Ser Leu Trp Arg Leu Glu Thr Leu Arg Val Ala Trp Ser Gly Ser His

275 280 285

Ile Arg Trp Gly Gln Pro Phe Arg Leu Arg His Val Thr Thr Gly Lys

290 295 300

Tyr Leu Ser Leu Met Glu Asp Lys Asn Leu Leu Leu Met Asp Lys Glu

305 310 315 320

Lys Ala Asp Val Lys Ser Thr Ala Phe Thr Phe Arg Ser Ser Lys Glu

325 330 335

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

340 345 350

Glu Ile Lys Tyr Gly Asp Ser Val Cys Tyr Ile Gln His Val Asp Thr

355 360 365

Gly Leu Trp Leu Thr Tyr Gln Ser Val Asp Val Lys Ser Val Arg Met

370 375 380

Gly Ser Ile Gln Arg Lys Ala Ile Met His His Glu Gly His Met Asp

385 390 395 400

Asp Gly Ile Ser Leu Ser Arg Ser Gln His Glu Glu Ser Leu Ile Ala

405 410 415

Arg Val Ile Arg Ser Thr Val Phe Leu Phe Asn Arg Phe Ile Arg Gly

420 425 430

Leu Asp Ala Leu Ser Lys Lys Ala Lys Ala Ser Thr Val Asp Leu Pro

435 440 445

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

450 455 460

Pro Pro Asp Glu His Leu Glu His Glu Asp Lys Gln Asn Arg Leu Arg

465 470 475 480

Ala Leu Lys Asn Arg Gln Asn Leu Phe Gln Glu Glu Gly Met Ile Asn

485 490 495

Leu Val Leu Glu Cys Ile Asp Arg Leu His Val Tyr Ser Ser Ala Ala

500 505 510

His Phe Ala Asp Val Ala Gly Arg Glu Ala Gly Glu Ser Trp Lys Ser

515 520 525

Ile Leu Asn Ser Leu Tyr Glu Leu Leu Ala Ala Leu Ile Arg Gly Asn

530 535 540

Arg Lys Asn Cys Ala Gln Phe Ser Gly Ser Leu Asp Trp Leu Ile Ser

545 550 555 560

Arg Leu Glu Arg Leu Glu Ala Ser Ser Gly Ile Leu Glu Val Leu His

565 570 575

Cys Val Leu Val Glu Ser Pro Glu Ala Leu Asn Ile Ile Lys Glu Gly

580 585 590

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

595 600 605

Lys Val Leu Asp Val Leu Cys Ser Leu Cys Val Cys His Gly Val Ala

610 615 620

Val Arg Ser Asn Gln His Leu Ile Cys Asp Asn Leu Leu Pro Gly Arg

625 630 635 640

Asp Leu Leu Leu Gln Thr Arg Leu Val Asn His Val Ser Ser Met Arg

645 650 655

Pro Asn Ile Phe Leu Gly Val Ser Glu Gly Ser Ala Gln Tyr Lys Lys

660 665 670

Trp Tyr Tyr Glu Leu Met Val Asp His Thr Glu Pro Phe Val Thr Ala

675 680 685

Glu Ala Thr His Leu Arg Val Gly Trp Ala Ser Thr Glu Gly Tyr Ser

690 695 700

Pro Tyr Pro Gly Gly Gly Glu Glu Trp Gly Gly Asn Gly Val Gly Asp

705 710 715 720

Asp Leu Phe Ser Tyr Gly Phe Asp Gly Leu His Leu Trp Ser Gly Cys

725 730 735

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

740 745 750

Asp Val Ile Ser Cys Cys Leu Asp Leu Ser Ala Pro Ser Ile Ser Phe

755 760 765

Arg Ile Asn Gly Gln Pro Val Gln Gly Met Phe Glu Asn Phe Asn Ile

770 775 780

Asp Gly Leu Phe Phe Pro Val Val Ser Phe Ser Ala Gly Ile Lys Val

785 790 795 800

Arg Phe Leu Leu Gly Gly Arg His Gly Glu Phe Lys Phe Leu Pro Pro

805 810 815

Pro Gly Tyr Ala Pro Cys Tyr Glu Ala Val Leu Pro Lys Glu Lys Leu

820 825 830

Lys Val Glu His Ser Arg Glu Tyr Lys Gln Glu Arg Thr Tyr Thr Arg

835 840 845

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

850 855 860

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

865 870 875 880

Ile Arg Glu Lys Leu Ala Glu Asn Ile His Glu Leu Trp Val Met Asn

885 890 895

Lys Ile Glu Leu Gly Trp Gln Tyr Gly Pro Val Arg Asp Asp Asn Lys

900 905 910

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

915 920 925

Arg Asn Tyr Asn Leu Gln Met Ser Leu Glu Thr Leu Lys Thr Leu Leu

930 935 940

Ala Leu Gly Cys His Val Gly Ile Ser Asp Glu His Ala Glu Asp Lys

945 950 955 960

Val Lys Lys Met Lys Leu Pro Lys Asn Tyr Gln Leu Thr Ser Gly Tyr

965 970 975

Lys Pro Ala Pro Met Asp Leu Ser Phe Ile Lys Leu Thr Pro Ser Gln

980 985 990

Glu Ala Met Val Asp Lys Leu Ala Glu Asn Ala His Asn Val Trp Ala

995 1000 1005

Arg Asp Arg Ile Arg Gln Gly Trp Thr Tyr Gly Ile Gln Gln Asp Val

1010 1015 1020

Lys Asn Arg Arg Asn Pro Arg Leu Val Pro Tyr Thr Leu Leu Asp Asp

1025 1030 1035 1040

Arg Thr Lys Lys Ser Asn Lys Asp Ser Leu Arg Glu Ala Val Arg Thr

1045 1050 1055

Leu Leu Gly Tyr Gly Tyr Asn Leu Glu Ala Pro Asp Gln Asp His Ala

1060 1065 1070

Ala Arg Ala Glu Val Cys Ser Gly Thr Gly Glu Arg Phe Arg Ile Phe

1075 1080 1085

Arg Ala Glu Lys Thr Tyr Ala Val Lys Ala Gly Arg Trp Tyr Phe Glu

1090 1095 1100

Phe Glu Thr Val Thr Ala Gly Asp Met Arg Val Gly Trp Ser Arg Pro

1105 1110 1115 1120

Gly Cys Gln Pro Asp Gln Glu Leu Gly Ser Asp Glu Arg Ala Phe Ala

1125 1130 1135

Phe Asp Gly Phe Lys Ala Gln Arg Trp His Gln Gly Asn Glu His Tyr

1140 1145 1150

Gly Arg Ser Trp Gln Ala Gly Asp Val Val Gly Cys Met Val Asp Met

1155 1160 1165

Asn Glu His Thr Met Met Phe Thr Leu Asn Gly Glu Ile Leu Leu Asp

1170 1175 1180

Asp Ser Gly Ser Glu Leu Ala Phe Lys Asp Phe Asp Val Gly Asp Gly

1185 1190 1195 1200

Phe Ile Pro Val Cys Ser Leu Gly Val Ala Gln Val Gly Arg Met Asn

1205 1210 1215

Phe Gly Lys Asp Val Ser Thr Leu Lys Tyr Phe Thr Ile Cys Gly Leu

1220 1225 1230

Gln Glu Gly Tyr Glu Pro Phe Ala Val Asn Thr Asn Arg Asp Ile Thr

1235 1240 1245

Met Trp Leu Ser Lys Arg Leu Pro Gln Phe Leu Gln Val Pro Ser Asn

1250 1255 1260

His Glu His Ile Glu Val Thr Arg Ile Asp Gly Thr Ile Asp Ser Ser

1265 1270 1275 1280

Pro Cys Leu Lys Val Thr Gln Lys Ser Phe Gly Ser Gln Asn Ser Asn

1285 1290 1295

Thr Asp Ile Met Phe Tyr Arg Leu Ser Met Pro Ile Glu Cys Ala Glu

1300 1305 1310

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

1315 1320 1325

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

1330 1335 1340

Val Leu Met Lys Thr Ala His Gly His Leu Val Pro Asp Arg Val Asp

1345 1350 1355 1360

Lys Asp Lys Glu Ala Thr Lys Pro Glu Phe Asn Asn His Lys Asp Tyr

1365 1370 1375

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

1380 1385 1390

Thr Lys Pro Asp Tyr Ser Thr Ser His Ser Ala Arg Leu Thr Glu Asp

1395 1400 1405

Val Leu Ala Asp Asp Arg Asp Asp Tyr Asp Phe Leu Met Gln Thr Ser

1410 1415 1420

Thr Tyr Tyr Tyr Ser Val Arg Ile Phe Pro Gly Gln Glu Pro Ala Asn

1425 1430 1435 1440

Val Trp Val Gly Trp Ile Thr Ser Asp Phe His Gln Tyr Asp Thr Gly

1445 1450 1455

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

1460 1465 1470

Lys Gly Lys Val His Glu Ser Ile Lys Arg Ser Asn Cys Tyr Met Val

1475 1480 1485

Cys Ala Gly Glu Ser Met Ser Pro Gly Gln Gly Arg Asn Asn Asn Gly

1490 1495 1500

Leu Glu Ile Gly Cys Val Val Asp Ala Ala Ser Gly Leu Leu Thr Phe

1505 1510 1515 1520

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

1525 1530 1535

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

1540 1545 1550

Phe Gln Phe Glu Leu Gly Arg Ile Lys Asn Val Met Pro Leu Ser Ala

1555 1560 1565

Gly Leu Phe Lys Ser Glu His Lys Asn Pro Val Pro Gln Cys Pro Pro

1570 1575 1580

Arg Leu His Val Gln Phe Leu Ser His Val Leu Trp Ser Arg Met Pro

1585 1590 1595 1600

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

1605 1610 1615

Trp Leu Val Gln Cys Leu Asp Pro Leu Gln Phe Met Ser Leu His Ile

1620 1625 1630

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

1635 1640 1645

Glu Leu Leu Lys Phe His Tyr His Thr Leu Arg Leu Tyr Ser Ala Val

1650 1655 1660

Cys Ala Leu Gly Asn His Arg Val Ala His Ala Leu Cys Ser His Val

1665 1670 1675 1680

Asp Glu Pro Gln Leu Leu Tyr Ala Ile Glu Asn Lys Tyr Met Pro Gly

1685 1690 1695

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

1700 1705 1710

Ser Tyr Ala Thr Ala Arg Leu Met Met Asn Asn Glu Tyr Ile Val Pro

1715 1720 1725

Met Thr Glu Glu Thr Lys Ser Ile Thr Leu Phe Pro Asp Glu Asn Lys

1730 1735 1740

Lys His Gly Leu Pro Gly Ile Gly Leu Ser Thr Ser Leu Arg Pro Arg

1745 1750 1755 1760

Met Gln Phe Ser Ser Pro Ser Phe Val Ser Ile Ser Asn Glu Cys Tyr

1765 1770 1775

Gln Tyr Ser Pro Glu Phe Pro Leu Asp Ile Leu Lys Ser Lys Thr Ile

1780 1785 1790

Gln Met Leu Thr Glu Ala Val Lys Glu Gly Ser Leu His Ala Arg Asp

1795 1800 1805

Pro Val Gly Gly Thr Thr Glu Phe Leu Phe Val Pro Leu Ile Lys Leu

1810 1815 1820

Phe Tyr Thr Leu Leu Ile Met Gly Ile Phe His Asn Glu Asp Leu Lys

1825 1830 1835 1840

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

1845 1850 1855

Pro Glu Glu Glu Ser Asp Thr Leu Glu Lys Glu Leu Ser Val Asp Asp

1860 1865 1870

Ala Lys Leu Gln Gly Ala Gly Glu Glu Glu Ala Lys Gly Gly Lys Arg

1875 1880 1885

Pro Lys Glu Gly Leu Leu Gln Met Lys Leu Pro Glu Pro Val Lys Leu

1890 1895 1900

Gln Met Cys Leu Leu Leu Gln Tyr Leu Cys Asp Cys Gln Val Arg His

1905 1910 1915 1920

Arg Ile Glu Ala Ile Val Ala Phe Ser Asp Asp Phe Val Ala Lys Leu

1925 1930 1935

Gln Asp Asn Gln Arg Phe Arg Tyr Asn Glu Val Met Gln Ala Leu Asn

1940 1945 1950

Met Ser Ala Ala Leu Thr Ala Arg Lys Thr Lys Glu Phe Arg Ser Pro

1955 1960 1965

Pro Gln Glu Gln Ile Asn Met Leu Leu Asn Phe Lys Asp Asp Lys Ser

1970 1975 1980

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

1985 1990 1995 2000

Glu Asp Leu Met Thr His Cys Gly Ile Glu Leu Asp Glu Asp Gly Ser

2005 2010 2015

Leu Asp Gly Asn Ser Asp Leu Thr Ile Arg Gly Arg Leu Leu Ser Leu

2020 2025 2030

Val Glu Lys Val Thr Tyr Leu Lys Lys Lys Gln Ala Glu Lys Pro Val

2035 2040 2045

Glu Ser Asp Ser Lys Lys Ser Ser Thr Leu Gln Gln Leu Ile Ser Glu

2050 2055 2060

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

2065 2070 2075 2080

Val Arg Ala Met Phe Val Leu Leu His Arg Gln Tyr Asp Gly Ile Gly

2085 2090 2095

Gly Leu Val Arg Ala Leu Pro Lys Thr Tyr Thr Ile Asn Gly Val Ser

2100 2105 2110

Val Glu Asp Thr Ile Asn Leu Leu Ala Ser Leu Gly Gln Ile Arg Ser

2115 2120 2125

Leu Leu Ser Val Arg Met Gly Lys Glu Glu Glu Lys Leu Met Ile Arg

2130 2135 2140

Gly Leu Gly Asp Ile Met Asn Asn Lys Val Phe Tyr Gln His Pro Asn

2145 2150 2155 2160

Leu Met Arg Ala Leu Gly Met His Glu Thr Val Met Glu Val Met Val

2165 2170 2175

Asn Val Leu Gly Gly Gly Glu Ser Lys Glu Ile Thr Phe Pro Lys Met

2180 2185 2190

Val Ala Asn Cys Cys Arg Phe Leu Cys Tyr Phe Cys Arg Ile Ser Arg

2195 2200 2205

Gln Asn Gln Lys Ala Met Phe Asp His Leu Ser Tyr Leu Leu Glu Asn

2210 2215 2220

Ser Ser Val Gly Leu Ala Ser Pro Ala Met Arg Gly Ser Thr Pro Leu

2225 2230 2235 2240

Asp Val Ala Ala Ala Ser Val Met Asp Asn Asn Glu Leu Ala Leu Ala

2245 2250 2255

Leu Arg Glu Pro Asp Leu Glu Lys Val Val Arg Tyr Leu Ala Gly Cys

2260 2265 2270

Gly Leu Gln Ser Cys Gln Met Leu Val Ser Lys Gly Tyr Pro Asp Ile

2275 2280 2285

Gly Trp Asn Pro Val Glu Gly Glu Arg Tyr Leu Asp Phe Leu Arg Phe

2290 2295 2300

Ala Val Phe Cys Asn Gly Glu Ser Val Glu Glu Asn Ala Asn Val Val

2305 2310 2315 2320

Val Arg Leu Leu Ile Arg Arg Pro Glu Cys Phe Gly Pro Ala Leu Arg

2325 2330 2335

Gly Glu Gly Gly Asn Gly Leu Leu Ala Ala Met Glu Glu Ala Ile Lys

2340 2345 2350

Ile Ala Glu Asp Pro Ser Arg Asp Gly Pro Ser Pro Asn Ser Gly Ser

2355 2360 2365

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

2370 2375 2380

Gly Asn Ala Ile Met Thr Phe Tyr Ser Ala Leu Ile Asp Leu Leu Gly

2385 2390 2395 2400

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

2405 2410 2415

Ile Arg Ile Arg Ser Ile Leu Arg Ser Leu Ile Pro Leu Gly Asp Leu

2420 2425 2430

Val Gly Val Ile Ser Ile Ala Phe Gln Met Pro Thr Ile Ala Lys Asp

2435 2440 2445

Gly Asn Val Val Glu Pro Asp Met Ser Ala Gly Phe Cys Pro Asp His

2450 2455 2460

Lys Ala Ala Met Val Leu Phe Leu Asp Arg Val Tyr Gly Ile Glu Val

2465 2470 2475 2480

Gln Asp Phe Leu Leu His Leu Leu Glu Val Gly Phe Leu Pro Asp Leu

2485 2490 2495

Arg Ala Ala Ala Ser Leu Asp Thr Ala Ala Leu Ser Ala Thr Asp Met

2500 2505 2510

Ala Leu Ala Leu Asn Arg Tyr Leu Cys Thr Ala Val Leu Pro Leu Leu

2515 2520 2525

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

2530 2535 2540

Ile Asp Ser Leu Leu His Thr Val Tyr Arg Leu Ser Lys Gly Cys Ser

2545 2550 2555 2560

Leu Thr Lys Ala Gln Arg Asp Ser Ile Glu Val Cys Leu Leu Ser Ile

2565 2570 2575

Cys Gly Gln Leu Arg Pro Ser Met Met Gln His Leu Leu Arg Arg Leu

2580 2585 2590

Val Phe Asp Val Pro Leu Leu Asn Glu His Ala Lys Met Pro Leu Lys

2595 2600 2605

Leu Leu Thr Asn His Tyr Glu Arg Cys Trp Lys Tyr Tyr Cys Leu Pro

2610 2615 2620

Gly Gly Trp Gly Asn Phe Gly Ala Ala Ser Glu Glu Glu Leu His Leu

2625 2630 2635 2640

Ser Arg Lys Leu Phe Trp Gly Ile Phe Asp Ala Leu Ser Gln Lys Lys

2645 2650 2655

Tyr Glu Gln Glu Leu Phe Lys Leu Ala Leu Pro Cys Leu Ser Ala Val

2660 2665 2670

Ala Gly Ala Leu Pro Pro Asp Tyr Met Glu Ser Asn Tyr Val Ser Met

2675 2680 2685

Met Glu Lys Gln Ser Ser Met Asp Ser Glu Gly Asn Phe Asn Pro Gln

2690 2695 2700

Pro Val Asp Thr Ser Asn Ile Thr Ile Pro Glu Lys Leu Glu Tyr Phe

2705 2710 2715 2720

Ile Asn Lys Tyr Ala Glu His Ser His Asp Lys Trp Ser Met Asp Lys

2725 2730 2735

Leu Ala Asn Gly Trp Ile Tyr Gly Glu Ile Tyr Ser Asp Ser Ser Lys

2740 2745 2750

Val Gln Pro Leu Met Lys Pro Tyr Lys Leu Leu Ser Glu Lys Glu Lys

2755 2760 2765

Glu Ile Tyr Arg Trp Pro Ile Lys Glu Ser Leu Lys Thr Met Leu Ala

2770 2775 2780

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

2785 2790 2795 2800

Tyr Asn Arg Thr Arg Arg Ile Ser Gln Thr Ser Gln Val Ser Val Asp

2805 2810 2815

Ala Ala His Gly Tyr Ser Pro Arg Ala Ile Asp Met Ser Asn Val Thr

2820 2825 2830

Leu Ser Arg Asp Leu His Ala Met Ala Glu Met Met Ala Glu Asn Tyr

2835 2840 2845

His Asn Ile Trp Ala Lys Lys Lys Lys Met Glu Leu Glu Ser Lys Gly

2850 2855 2860

Gly Gly Asn His Pro Leu Leu Val Pro Tyr Asp Thr Leu Thr Ala Lys

2865 2870 2875 2880

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

2885 2890 2895

Gln Ile Asn Gly Tyr Ala Val Ser Arg Gly Phe Lys Asp Leu Glu Leu

2900 2905 2910

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

2915 2920 2925

Leu Ile Arg Tyr Val Asp Glu Ala His Gln Tyr Ile Leu Glu Phe Asp

2930 2935 2940

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

2945 2950 2955 2960

Lys Phe Phe Ala Lys Val Val Leu Pro Leu Ile Asp Gln Tyr Phe Lys

2965 2970 2975

Asn His Arg Leu Tyr Phe Leu Ser Ala Ala Ser Arg Pro Leu Cys Ser

2980 2985 2990

Gly Gly His Ala Ser Asn Lys Glu Lys Glu Met Val Thr Ser Leu Phe

2995 3000 3005

Cys Lys Leu Gly Val Leu Val Arg His Arg Ile Ser Leu Phe Gly Asn

3010 3015 3020

Asp Ala Thr Ser Ile Val Asn Cys Leu His Ile Leu Gly Gln Thr Leu

3025 3030 3035 3040

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

3045 3050 3055

Leu Arg Ala Phe Leu Asp Asn Ala Ala Glu Asp Leu Glu Lys Thr Met

3060 3065 3070

Glu Asn Leu Lys Gln Gly Gln Phe Thr His Thr Arg Asn Gln Pro Lys

3075 3080 3085

Gly Val Thr Gln Ile Ile Asn Tyr Thr Thr Val Ala Leu Leu Pro Met

3090 3095 3100

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

3105 3110 3115 3120

Leu Ile Leu Glu Asp Val Gln Val Ser Cys Tyr Arg Ile Leu Thr Ser

3125 3130 3135

Leu Tyr Ala Leu Gly Thr Ser Lys Ser Ile Tyr Val Glu Arg Gln Arg

3140 3145 3150

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

3155 3160 3165

Ala Phe Leu Glu Thr His Leu Asp Lys His Asn Ile Tyr Ser Ile Tyr

3170 3175 3180

Asn Thr Lys Ser Ser Arg Glu Arg Ala Ala Leu Ser Leu Pro Thr Asn

3185 3190 3195 3200

Val Glu Asp Val Cys Pro Asn Ile Pro Ser Leu Glu Lys Leu Met Glu

3205 3210 3215

Glu Ile Val Glu Leu Ala Glu Ser Gly Ile Arg Tyr Thr Gln Met Pro

3220 3225 3230

His Val Met Glu Val Ile Leu Pro Met Leu Cys Ser Tyr Met Ser Arg

3235 3240 3245

Trp Trp Glu His Gly Pro Glu Asn Asn Pro Glu Arg Ala Glu Met Cys

3250 3255 3260

Cys Thr Ala Leu Asn Ser Glu His Met Asn Thr Leu Leu Gly Asn Ile

3265 3270 3275 3280

Leu Lys Ile Ile Tyr Asn Asn Leu Gly Ile Asp Glu Gly Ala Trp Met

3285 3290 3295

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

3300 3305 3310

Gln Leu Leu Lys Thr His Phe Leu Pro Leu Met Glu Lys Leu Lys Lys

3315 3320 3325

Lys Ala Ala Thr Val Val Ser Glu Glu Asp His Leu Lys Ala Glu Ala

3330 3335 3340

Arg Gly Asp Met Ser Glu Ala Glu Leu Leu Ile Leu Asp Glu Phe Thr

3345 3350 3355 3360

Thr Leu Ala Arg Asp Leu Tyr Ala Phe Tyr Pro Leu Leu Ile Arg Phe

3365 3370 3375

Val Asp Tyr Asn Arg Ala Lys Trp Leu Lys Glu Pro Asn Pro Glu Ala

3380 3385 3390

Glu Glu Leu Phe Arg Met Val Ala Glu Val Phe Ile Tyr Trp Ser Lys

3395 3400 3405

Ser His Asn Phe Lys Arg Glu Glu Gln Asn Phe Val Val Gln Asn Glu

3410 3415 3420

Ile Asn Asn Met Ser Phe Leu Ile Thr Asp Thr Lys Ser Lys Met Ser

3425 3430 3435 3440

Lys Ala Ala Val Ser Asp Gln Glu Arg Lys Lys Met Lys Arg Lys Gly

3445 3450 3455

Asp Arg Tyr Ser Met Gln Thr Ser Leu Ile Val Ala Ala Leu Lys Arg

3460 3465 3470

Leu Leu Pro Ile Gly Leu Asn Ile Cys Ala Pro Gly Asp Gln Glu Leu

3475 3480 3485

Ile Ala Leu Ala Lys Asn Arg Phe Ser Leu Lys Asp Thr Glu Asp Glu

3490 3495 3500

Val Arg Asp Ile Ile Arg Ser Asn Ile His Leu Gln Gly Lys Leu Glu

3505 3510 3515 3520

Asp Pro Ala Ile Arg Trp Gln Met Ala Leu Tyr Lys Asp Leu Pro Asn

3525 3530 3535

Arg Thr Asp Asp Thr Ser Asp Pro Glu Lys Thr Val Glu Arg Val Leu

3540 3545 3550

Asp Ile Ala Asn Val Leu Phe His Leu Glu Gln Lys Ser Lys Arg Val

3555 3560 3565

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

3570 3575 3580

Ala Val Trp His Lys Leu Leu Ser Lys Gln Arg Lys Arg Ala Val Val

3585 3590 3595 3600

Ala Cys Phe Arg Met Ala Pro Leu Tyr Asn Leu Pro Arg His Arg Ala

3605 3610 3615

Val Asn Leu Phe Leu Gln Gly Tyr Glu Lys Ser Trp Ile Glu Thr Glu

3620 3625 3630

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

3635 3640 3645

Ala Glu Pro Pro Glu Glu Asp Glu Gly Thr Lys Arg Val Asp Pro Leu

3650 3655 3660

His Gln Leu Ile Leu Leu Phe Ser Arg Thr Ala Leu Thr Glu Lys Cys

3665 3670 3675 3680

Lys Leu Glu Glu Asp Phe Leu Tyr Met Ala Tyr Ala Asp Ile Met Ala

3685 3690 3695

Lys Ser Cys His Asp Glu Glu Asp Asp Asp Gly Glu Glu Glu Val Lys

3700 3705 3710

Ser Phe Glu Glu Lys Glu Met Glu Lys Gln Lys Leu Leu Tyr Gln Gln

3715 3720 3725

Ala Arg Leu His Asp Arg Gly Ala Ala Glu Met Val Leu Gln Thr Ile

3730 3735 3740

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

3745 3750 3755 3760

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

3765 3770 3775

Met Leu Asp Tyr Leu Lys Glu Lys Lys Asp Val Gly Phe Phe Gln Ser

3780 3785 3790

Leu Ala Gly Leu Met Gln Ser Cys Ser Val Leu Asp Leu Asn Ala Phe

3795 3800 3805

Glu Arg Gln Asn Lys Ala Glu Gly Leu Gly Met Val Thr Glu Glu Gly

3810 3815 3820

Ser Gly Glu Lys Val Leu Gln Asp Asp Glu Phe Thr Cys Asp Leu Phe

3825 3830 3835 3840

Arg Phe Leu Gln Leu Leu Cys Glu Gly His Asn Ser Asp Phe Gln Asn

3845 3850 3855

Tyr Leu Arg Thr Gln Thr Gly Asn Asn Thr Thr Val Asn Ile Ile Ile

3860 3865 3870

Ser Thr Val Asp Tyr Leu Leu Arg Val Gln Glu Ser Ile Ser Asp Phe

3875 3880 3885

Tyr Trp Tyr Tyr Ser Gly Lys Asp Val Ile Asp Glu Gln Gly Gln Arg

3890 3895 3900

Asn Phe Ser Lys Ala Ile Gln Val Ala Lys Gln Val Phe Asn Thr Leu

3905 3910 3915 3920

Thr Glu Tyr Ile Gln Gly Pro Cys Thr Gly Asn Gln Gln Ser Leu Ala

3925 3930 3935

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

3940 3945 3950

His Met Gln Met Lys Leu Ser Gln Asp Ser Ser Gln Ile Glu Leu Leu

3955 3960 3965

Lys Glu Leu Met Asp Leu Gln Lys Asp Met Val Val Met Leu Leu Ser

3970 3975 3980

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

3985 3990 3995 4000

Asp Met Leu Val Glu Ser Ser Asn Asn Val Glu Met Ile Leu Lys Phe

4005 4010 4015

Phe Asp Met Phe Leu Lys Leu Lys Asp Leu Thr Ser Ser Asp Thr Phe

4020 4025 4030

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

4035 4040 4045

His Lys Ala Met Glu Ser His Lys His Tyr Thr Gln Ser Glu Thr Glu

4050 4055 4060

Phe Leu Leu Ser Cys Ala Glu Thr Asp Glu Asn Glu Thr Leu Asp Tyr

4065 4070 4075 4080

Glu Glu Phe Val Lys Arg Phe His Glu Pro Ala Lys Asp Ile Gly Phe

4085 4090 4095

Asn Val Ala Val Leu Leu Thr Asn Leu Ser Glu His Met Pro Asn Asp

4100 4105 4110

Thr Arg Leu Gln Thr Phe Leu Glu Leu Ala Glu Ser Val Leu Asn Tyr

4115 4120 4125

Phe Gln Pro Phe Leu Gly Arg Ile Glu Ile Met Gly Ser Ala Lys Arg

4130 4135 4140

Ile Glu Arg Val Tyr Phe Glu Ile Ser Glu Ser Ser Arg Thr Gln Trp

4145 4150 4155 4160

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

4165 4170 4175

Val Asn Glu Gly Gly Glu Lys Glu Lys Met Glu Leu Phe Val Asn Phe

4180 4185 4190

Cys Glu Asp Thr Ile Phe Glu Met Gln Leu Ala Ala Gln Ile Ser Glu

4195 4200 4205

Ser Asp Leu Asn Glu Arg Ser Ala Asn Lys Glu Glu Ser Glu Lys Glu

4210 4215 4220

Arg Pro Glu Glu Gln Gly Pro Arg Met Ala Phe Phe Ser Ile Leu Thr

4225 4230 4235 4240

Val Arg Ser Ala Leu Phe Ala Leu Arg Tyr Asn Ile Leu Thr Leu Met

4245 4250 4255

Arg Met Leu Ser Leu Lys Ser Leu Lys Lys Gln Met Lys Lys Val Lys

4260 4265 4270

Lys Met Thr Val Lys Asp Met Val Thr Ala Phe Phe Ser Ser Tyr Trp

4275 4280 4285

Ser Ile Phe Met Thr Leu Leu His Phe Val Ala Ser Val Phe Arg Gly

4290 4295 4300

Phe Phe Arg Ile Ile Cys Ser Leu Leu Leu Gly Gly Ser Leu Val Glu

4305 4310 4315 4320

Gly Ala Lys Lys Ile Lys Val Ala Glu Leu Leu Ala Asn Met Pro Asp

4325 4330 4335

Pro Thr Gln Asp Glu Val Arg Gly Asp Gly Glu Glu Gly Glu Arg Lys

4340 4345 4350

Pro Leu Glu Ala Ala Leu Pro Ser Glu Asp Leu Thr Asp Leu Lys Glu

4355 4360 4365

Leu Thr Glu Glu Ser Asp Leu Leu Ser Asp Ile Phe Gly Leu Asp Leu

4370 4375 4380

Lys Arg Glu Gly Gly Gln Tyr Lys Leu Ile Pro His Asn Pro Asn Ala

4385 4390 4395 4400

Gly Leu Ser Asp Leu Met Ser Asn Pro Val Pro Met Pro Glu Val Gln

4405 4410 4415

Glu Lys Phe Gln Glu Gln Lys Ala Lys Glu Glu Glu Lys Glu Glu Lys

4420 4425 4430

Glu Glu Thr Lys Ser Glu Pro Glu Lys Ala Glu Gly Glu Asp Gly Glu

4435 4440 4445

Lys Glu Glu Lys Ala Lys Glu Asp Lys Gly Lys Gln Lys Leu Arg Gln

4450 4455 4460

Leu His Thr His Arg Tyr Gly Glu Pro Glu Val Pro Glu Ser Ala Phe

4465 4470 4475 4480

Trp Lys Lys Ile Ile Ala Tyr Gln Gln Lys Leu Leu Asn Tyr Phe Ala

4485 4490 4495

Arg Asn Phe Tyr Asn Met Arg Met Leu Ala Leu Phe Val Ala Phe Ala

4500 4505 4510

Ile Asn Phe Ile Leu Leu Phe Tyr Lys Val Ser Thr Ser Ser Val Val

4515 4520 4525

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

4530 4535 4540

Thr Ser Leu Asp Ser Ser Ser His Arg Ile Ile Ala Val His Tyr Val

4545 4550 4555 4560

Leu Glu Glu Ser Ser Gly Tyr Met Glu Pro Thr Leu Arg Ile Leu Ala

4565 4570 4575

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

4580 4585 4590

Leu Lys Val Pro Leu Val Ile Phe Lys Arg Glu Lys Glu Val Ala Arg

4595 4600 4605

Lys Leu Glu Phe Asp Gly Leu Tyr Ile Thr Glu Gln Pro Ser Glu Asp

4610 4615 4620

Asp Ile Lys Gly Gln Trp Asp Arg Leu Val Ile Asn Thr Gln Ser Phe

4625 4630 4635 4640

Pro Asn Asn Tyr Trp Asp Lys Phe Val Lys Arg Lys Val Met Asp Lys

4645 4650 4655

Tyr Gly Glu Phe Tyr Gly Arg Asp Arg Ile Ser Glu Leu Leu Gly Met

4660 4665 4670

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

4675 4680 4685

Lys Lys Asp Ser Ser Leu Ser Ala Val Leu Asn Ser Ile Asp Val Lys

4690 4695 4700

Tyr Gln Met Trp Lys Leu Gly Val Val Phe Thr Asp Asn Ser Phe Leu

4705 4710 4715 4720

Tyr Leu Ala Trp Tyr Met Thr Met Ser Val Leu Gly His Tyr Asn Asn

4725 4730 4735

Phe Phe Phe Ala Ala His Leu Leu Asp Ile Ala Met Gly Phe Lys Thr

4740 4745 4750

Leu Arg Thr Ile Leu Ser Ser Val Thr His Asn Gly Lys Gln Leu Val

4755 4760 4765

Leu Thr Val Gly Leu Leu Ala Val Val Val Tyr Leu Tyr Thr Val Val

4770 4775 4780

Ala Phe Asn Phe Phe Arg Lys Phe Tyr Asn Lys Ser Glu Asp Gly Asp

4785 4790 4795 4800

Thr Pro Asp Met Lys Cys Asp Asp Met Leu Thr Cys Tyr Met Phe His

4805 4810 4815

Met Tyr Val Gly Val Arg Ala Gly Gly Gly Ile Gly Asp Glu Ile Glu

4820 4825 4830

Asp Pro Ala Gly Asp Glu Tyr Glu Ile Tyr Arg Ile Ile Phe Asp Ile

4835 4840 4845

Thr Phe Phe Phe Phe Val Ile Val Ile Leu Leu Ala Ile Ile Gln Gly

4850 4855 4860

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

4865 4870 4875 4880

Lys Glu Asp Met Glu Thr Lys Cys Phe Ile Cys Gly Ile Gly Asn Asp

4885 4890 4895

Tyr Phe Asp Thr Val Pro His Gly Phe Glu Thr His Thr Leu Gln Glu

4900 4905 4910

His Asn Leu Ala Asn Tyr Leu Phe Phe Leu Met Tyr Leu Ile Asn Lys

4915 4920 4925

Asp Glu Thr Glu His Thr Gly Gln Glu Ser Tyr Val Trp Lys Met Tyr

4930 4935 4940

Gln Glu Arg Cys Trp Glu Phe Phe Pro Ala Gly Asp Cys Phe Arg Lys

4945 4950 4955 4960

Gln Tyr Glu Asp Gln Leu Asn

4965

Claims

1.与遗传性心律失常有关的突变基因，其特征在于，与野生型RYR2基因编码DNA的参考序列相比，突变基因RYR2的核苷酸序列为SEQ ID NO:3；在基因组位置chr1:237608771 处，碱基G突变为碱基T，在基因组位置chr1:237608774处，碱基C突变为碱基T；参考基因组版本是GRCh37。

2.根据权利要求1所述的与遗传性心律失常有关的突变基因在制备检测试剂盒中的应用。

3.根据权利要求2所述的应用，其特征在于，所述检测试剂盒包括引物SEQ ID NO:1和SEQ ID NO:2。

4.根据权利要求3所述的应用，其特征在于，所述检测试剂盒还包括Taq DNA聚合酶和PCR缓冲液。