专利名称：一种骨骼肌和心肌功能相关基因及其编码的蛋白质的制作方法
技术领域：
本发明涉及一种与骨骼肌和心肌功能相关的基因及其编码的蛋白质，特别是涉及一种可能与扩张型心肌病相关的基因及其编码的蛋白质。
背景技术：
骨骼肌和心肌是人体最重要的组织之一，功能是收缩、拉动身体各部位，使其完成相应的功能。肌肉组织的发育及其行使功能由多种基因控制，基因功能失常则引发相应的肌病。目前肌萎缩、肌无力和心肌病仍是严重威胁人类健康的疾病，寻找早期诊断和治疗的生物学指标势在必行，骨骼肌和心肌功能相关基因的发现提供了潜在的靶点。
据估计在人类基因组中大约有5000个潜在的药物靶点，迄今为止成功应用于药物开发的人类药物靶标约有500种，仅占潜在药物靶点的十分之一，因此该领域具有巨大的开发潜力。
发明内容
本发明的目的是提供一种与骨骼肌和心肌功能相关的基因及其编码的蛋白质。
一种与骨骼肌和心肌功能相关的基因，它是下列核苷酸序列之一1)序列表中序列1的DNA序列；2)与序列表中序列1限定的DNA序列具有90％以上同源性，且编码相同功能蛋白质的DNA序列。
所述与骨骼肌和心肌功能相关基因的完整开放读码框为序列表中序列1的第166至5253共5088个碱基。
一种与骨骼肌和心肌功能相关的蛋白质，它具有序列表中序列2的氨基酸序列或将序列2的氨基酸序列经过一个或几个氨基酸残基的取代、缺失或添加且具有与序列2的氨基酸序列相同活性的由序列2衍生的蛋白质。
利用NCBI的Genbank数据库对本发明基因进行检索，结果表明数据库中没有基因与此基因序列完全相同。进一步应用GDB的数据库检索，结果表明，本发明基因应定位于10号染色体的长臂25至26区(10q25-q26)。
本发明所提供的基因在小鼠中有对应的同源物。该同源物特异表达于骨骼肌和心肌组织，被认为在肌纤维生成过程中起作用，是肌纤维组装的引发中心；该同源物在MLP敲除和tropomodulin转基因小鼠模型中的表达都上调，提示有可能作为扩张型心肌病(dilated cardiomyopathy，DCM)的早期标志。本发明所提供的基因在人骨骼肌、心肌和脑中都表达，生物信息学分析和亚细胞定位结果表明，本发明所提供的基因与其同源物有相同的功能。因此本发明的基因及其编码的蛋白质有望为临床扩张型心肌病的诊断和治疗提供有意义的生物学指标、药物靶标或蛋白类药物，将会带来巨大的社会效益及经济效益，而且，对于基因不同研究阶段所开发的不同产品，包括新抗体、生物芯片的靶基因、筛选出的新药等，也将促进形成新兴的人类基因产业。
图1为本发明基因在几种组织中表达的检测结果。
图2为本发明基因与小鼠中对应同源物结构域分析结果。
图3为本发明基因表达的亚细胞定位结果。
具体实施例方式
实施例1、本发明基因的克隆本发明的基因cDNA克隆自人骨骼肌cDNA文库。根据生物信息学分析结果，设计2对引物上游引物P1(ATGAATGTGCAGCCCTGTTC)、P2(TCATCGACCCTCTGCCG)和下游引物P3(AAGACTCAGTGGGCCAAGAA)、P4(CGAAGATTACGCCAGGACTC)，以人骨骼肌cDNA文库(Clontech)为模板，用高保真DNA聚合酶Pyrobest(TaKaRa)扩增出对应片段。上游片段扩增采用Touchdown PCR技术，条件如下94℃预变性3min；94℃变性15s，60℃退火15s，每经过一个循环退火温度降低0.5℃，72℃延伸3min，共25个循环；94℃变性15s，55℃退火15s，72℃延伸3min，共10个循环；72℃ 5min。PCR结束后，补加1μl rTaq多聚酶(TaKaRa)72℃ 20min加尾。下游片段扩增条件如下94℃预变性3min；94℃变性15s，55℃退火15s，72℃延伸3min，共38个循环；72℃ 5min。PCR结束后，补加1μl rTaq多聚酶(TaKaRa)72℃ 20min加尾。0.7％琼脂糖凝胶电泳分离PCR产物，采用玻璃奶回收试剂盒(北京博大泰克生物基因技术有限责任公司)从琼脂糖凝胶中回收目的片段。目的片段以3∶1摩尔比与pMD18-T载体(TaKaRa)进行连接。将连接产物转化大肠杆菌JM109，挑选白色菌落进行PCR和酶切鉴定，阳性克隆送上海博亚生物技术有限公司测序。2个片段拼接后用ESTBlast程序(http//www.hgmp.mrc.ac.uk/ESTBlast)进行电子延伸，将延伸后的序列与人类基因组数据库作BLAST分析(http//www.ncbi.nlm.nih.gov/BLAST)，确认其可靠性。根据拼接后的序列，设计2对引物外引物P5(TCAGCAAAGGAAAGTGGAAG)、P6(GATCTGGGATGGGCTGG)和内引物P7(GAATTCGGATGAATGTGCAGCCCTGTTC)、P8(CTCGAGTCACAACAGCAGGGCCTTCTTC)，内引物上分别添加EcoRI和XhoI酶切位点，以便进行后续操作。以人骨骼肌cDNA文库(Clontech)为模板，用Advantage II DNA聚合酶(Clontech)和外引物P5、P6进行第一轮PCR，条件如下94℃预变性3min；94℃变性30s，55℃退火30s，72℃延伸6min，共20个循环；72℃ 10min。将PCR产物用去离子水稀释100倍，取1μl作模板，用内引物P7和P8进行第二轮PCR，条件与第一轮PCR相同。电泳分离PCR产物，回收后连入pMD18-T载体，挑选阳性克隆测序。结果表明本发明所克隆到的基因为序列表中序列1的DNA序列。
实施例2、本发明基因在几种成人组织中表达的检测用RT-PCR法。分别以成人脑、骨髓、骨骼肌和心脏组织的cDNA文库(Clontech)为模板，用引物P5、P6、P7和P8进行巢式PCR，扩增本发明基因编码区5088bp的片段。回收特异性片段后连入pMD18-T载体测序证实。结果如图1所示，1为骨髓，2为心脏，3为脑，4为相对分子质量标记DL15000，5为骨骼肌。除骨髓组织外，从脑、骨骼肌和心脏组织中均扩增到5.1kb片段。
实施例3、本发明基因的结构域分析在ExPASy分子生物学服务器(http//cn.expasy.org)上进行。结果如图2，A为本发明基因的结构域组成，B为小鼠中对应同源物的结构域组成。可以看出，二者是极其相似的，提示有相同的功能。
实施例4、本发明基因表达的亚细胞定位回收本发明基因编码区，以相同读码框架连入pCMV-myc载体，构建融合表达载体。用脂质体Lipofactamine 2000(Gibco)介导的方法，将融合表达载体瞬时转染约80％汇合的COS7细胞，使本发明基因与c-myc标签融合表达。转染24~48h后依次用4％多聚甲醛固定细胞30min，用0.5％Triton X-100处理15min，用山羊血清封闭10min，用小鼠抗人c-myc单克隆抗体(北京中山生物技术有限公司)37℃孵育1h，用红色荧光TRITC标记的山羊抗小鼠IgG(北京中山生物技术有限公司)37℃孵育30min，用50mg/L4’，6-二乙酰基-2-苯基吲哚(DAPI)(Sigma)复染15min，其间均用PBS洗涤3次，每次5min。荧光显微镜下观察红色荧光和蓝色荧光的分布。以转染pCMV-myc载体的COS7细胞和未转染基因的COS7细胞作为对照。结果见图3A为转染融合表达载体的COS7细胞，B为转染pCMV-myc的COS7细胞，C为DAPI显示转染融合表达载体的COS7细胞的细胞核(箭头所示)，D为DAPI显示转染pCMV-myc的COS7细胞的细胞核(箭头所示)。从图中可以看出，本发明的基因表达于胞浆，这与其可能行使的参与肌纤维生成的功能是相吻合的。
序列表<160>2<210>1<211>5417<212>DNA<213>人属人种(Homo sapiens)<400>1aaaactagaa ggtcagcaaa ggaaagtgga agttggattc tgaaagatcg 50aggtgcccat aggaatttta tggtcgtcgg attttgaaga cttgaactag 100actgggggtt ctccttgcat ttcttgcctg ttgcctatct ttgtcctctc 150tcttccggct tcgagatgaa tgtgcagccc tgttctaggt gtgggtatgg 200ggtttatcct gccgagaaga tcagctgtat agatcagata tggcataaag 250cctgttttca ctgtgaagtt tgcaagatga tgctgtctgt taataacttt 300gtgagtcacc agaaaaagcc gtactgtcac gcccataacc ctaagaacaa 350cactttcacc agtgtctatc acactccatt aaatctaaat gtgaggacat 400ttccagaggc catcagtggg atccatgacc aagaagatgg tgaacagtgt 450aaatcagttt ttcattggga catgaaatcc aaggataagg aaggtgcacc 500taacaggcag ccactggcaa atgagagagc ctattggact ggatatgggg 550aagggaatgc ttggtgccca ggagctctgc cagaccccga aattgtaagg 600atggttgagg ctcgaaagtc tcttggtgag gaatatacag aagactatga 650gcaacccagg ggcaagggga gctttccagc catgatcaca cctgcttatc 700aaagggccaa gaaagccaac cagctggcca gccaagtgga gtataagaga 750gggcatgatg aacgcatctc caggttctcc acggtggcgg atactcctga 800gctgctacgg agcaaggctg gggcacagct tcaaagtgat gtgagataca 850cagaggacta tgaacaacaa agagggaaag gcagtttccc tgcgatgatc 900acacccgcct atcagatagc caaaagagcc aatgagctgg caagtgatgt 950gaggtaccat caacaatatc aaaaagaaat gaggggaatg gctggtccag 1000ccattggagc tgagggcatc ttgacaaggg aatgtgcaga ccaatatggc 1050cagggttacc cggaggagta tgaggagcac aggggaaagg gcagcttccc 1100agctatgatc actccagcat atcagaacgc caagaaagct cacgaactcg 1150ctagtgacat aaaatacagg caggacttca ataagatgaa aggcactgca 1200cattatcact cgcttccagc tcaagacaac ttggttctca aacgggctca 1250gagcgtaaac aaactcgtga gtgagaataa atataaagaa aactaccaga 1300accacatgag aggccgctat gaaggagttg gtatggacag acgcactctg 1350catgctatga aagttggcag cctggcaagc aacgttgcct acaaagctga 1400ttataaacat gatattgtcg actacaacta cccagccact ctcacgcctt 1450cctatcaaac agctatgaaa ctggtgccct tgaaagatgc caattatagg 1500cagagcatcg acaagttgaa gtacagctcg gtgactgaca ccccacagat 1550tgttcaagcc aaaatcaatg cccagcagct gagtcatgtg aattaccgtg 1600ctgactatga gaaaaataag ttgaattaca cattgcccca ggatgttcct 1650cagctggtga aggccaaaac caatgccaaa ctcttcagtg aggttaagta 1700taaagaaggc tgggagaaga caaaggggaa aggatttgag atgaagctgg 1750atgccatgtc tctgctggcc gccaaagcct ctggggagct tgctagcaat 1800attaaatata aagaagaata tgaaaagaca aaaggcaaag ccatgggaac 1850agccgactct aggcttctgc actccctgca gattgctaag atgagcagtg 1900
aggttgaata taagaaaggc tttgaagaga gtaagaccog gtttcacctg1950cccatggata tggtaaacat caggcatgct aagaaggccc aaactctcgc2000cagtgacctg gactacagga agaaactgca tgaatacact gtgctgcctg2050aagatatgaa gactcagtgg gccaagaagg tctatgggct tcagagcgag2100ctgcagtaca aggctgacct ggcatggatg aagggagttg ggtggctgac2150agaggggagt ctcaacttgg aacaagccaa gaaggctgga cagctggtca2200gcgagaaaaa ctaccggcag agggtcgatg agctgaagtt caccagcgtg2250accgacagct cccagatgga gcacgccaag aagagccagg agctacagag2300cggggtggcc tacaaggcag gaaatgagca gtctgtccat cagtatacca2350tcagcaaaga cgagcctctc ttcctgcagg cccgagccaa tgctgcaaat2400ctcagcgaga aactgtataa gagcagctgg gaaaaccaga aagcaaaagg2450gtttgagctg cgtcttgatt ccttgacctt cctggcagcc aaagccaaga2500gggacctggc tagcgaggtg aagtacaagg aggattatga gagatccaga2550gggaagctca ttggggcaaa agatgtacag ggagattcgc aaatgagcca2600ctcactgcaa atgtccaagc tgcagagtga gctggagtac aagaagggat2650tcgaggacac caaatcccaa tgccacgtct ccctggacat ggtccacctc2700gtgcatgccc gcaaagctca gcatttagcc acagacgtag gctacaagac2750agcggaacat cactttacgg ctttgcccac agacatgaag gtggaatggg2800ccaagaaggc ttatggctta cagagtgata accaatacag ggcagatgtg2850aagtggatga aaggcatggg ctgggtcgcc accgggtcat taaatgtgga2900gcaggcgaag aaggcaggag aactcattag cgagaagaag taccgtcagc2950atccagatgc tttgaagttt accagtatta aagacactcc ggagatggtc3000caggccagaa ttagttatac ccaagcagtg gatagattat acagggaaca3050aggagaaaac ataaagcatc attacacacc gactgctgac ctccctgaag3100tcctgctggc caagctgaat gccatgaata tcagtgagac gcgttataag3150gaatcctgga gcaaacttcg agatggtggc tataaactga ggttggatgc3200ccttccattc caagcagcaa aggcttctgg tgaaatcata agtgattaca3250aatacaaaga ggcatttgag aaaatgaaag gacagatgct tggttcccgg3300agcttggaag atgatatcag ccttgcacat tcagtgtatg cgacctcact3350gcagagtgat gtgaattaca agaaaggctt tgaacactca aaggcgcagt3400tccatctgcc gttggacatg gccgccctgg tgcatgccaa gaaggctcag3450accctggcca gcaatcagga ctacaaacat ccactgcccc agtacacttc3500cttggcagaa gacctgaggc tgagctgtgc caagaaagct cacaaattgc3550agagtgagaa tttgtaccgg tcagacctga actttatgcg aggtgttgca3600tgtgtcattc caggcacgtt agagattgaa gggaggaaga aagcatcgga3650actcatcagt gagagtaaat atcggcagca tccccactca ttcaagtaca3700cagctgtgac agacactccc aacctccttc atgcgaaatt cagcaaccag3750ataacgaatg agcgcctcta taaagcagct ggagaggatg caagacacga3800gtatacaatg accctgggtc tgcccgagtt catccgagca aaaacgaatg3850cagccaacct gagtgacgca agatacaaag agtcctggcg taatcttcgt3900gctcaaggct acaagctgac aatagaagcg ctccccttcc aggctgcccg3950ggcctctgga gatatagcca gtgattttct ctacagacat gactttgtga4000aggagcgagg gaaactcata gggccccaga gtgtaagaga cgacccccgg4050atccagcact gccggcgcat gggccagctg cagagcgagc ttcagtacag4100gaggggggcg accagcagcc aagcccagtt ccatctgccc atggacatgg4150tgcacctggt ccatgccaag aatgcccagg ctctggccag cgaccacgac4200tacaggacac agtatcacaa gttcacagca ctgcccgagg acctgaagat4250ggcctgggcc aagaaagccc atgccctgca gagtgagttg cgctacaagt4300
cagacctgat cggcatgaag ggcataggat ggctggcgct gagotcccca 4350cagatggaga gtgcaaagaa ggctggagaa ctcatcagcg agaccaagta 4400ccgtaaaaaa ccagacagta tcaagttcac cacagtggtt gactccccag 4450acctggttca tgccaagaac agctatatgc actgcaatga gcgcatgtat 4500agatctggag atgcagaatc cctgcacaga tacaccctga ttcccgacca 4550tcccgatttc acccgagctc gcctcaatgc gctgcatctg agtgacaaag 4600tctacagaaa ctcctgggag cagacccggg ctggcagtta tgacttcagg 4650ctggatgcca tccccttcca gactgcccgg gcatctaggg agatcgccag 4700tgatttccgg tacaaagagg ctttcctgcg ggaccgaggc ctgcagatcg 4750ggtaccgcag tgtcgacgat gacccaagga tgaagcattt cctcaacgtt 4800ggcaggctcc agagtgacaa tgagtacaag aaggactttg ccaagagtcg 4850gtcccagttt cacagcagca cagaccagcc cggcctcctt caggccaaga 4900ggagccagca gctggccagt gatgtgcact acaggcagcc cctgccccag 4950cccacctgcg acccggagca gctgggcctc aggcatgctc agaaggccca 5000ccagctgcag agtgatgtca agtataaatc agacttgaac ctgaccagag 5050gtgttggctg gacccctcct ggctcctaca aagtggaaat ggctcggcgg 5100gctgcggaac tggcca cgc aaggggcctg ggtctccagg gagcttaccg 5150gggggcagaa gcagtggagg ctggagatca tcagagtggg gaggtgaacc 5200cagatgccac tgagattctg cacgt aaaa agaagaaggc cctgctgttg 5250tgagccatgt ccaccctgat tcctgagagg ccagagagga agtttgttca 5300ccagagacag gcttcagatg gctttgattt cggcaagctg gaatggccca 5350ccagcccatc ccagatcttc ctttattaaa ataataactc tgaaagcaga 5400tggaaaaaaa aaaaaaa 5417<210>2<211>1695<212>PRT<213>人属人种(Homo sapiens)<400>2Met Asn Val Gln Pro Cys Ser Arg Cys Gly Tyr Gly Val Tyr Pro15 10 15Ala Glu Lys Ile Ser Cys Ile Asp Gln Ile Trp His Lys Ala Cys20 25 30Phe His Cys Glu Val Cys Lys Met Met Leu Ser Val Asn Asn Phe35 40 45Val Ser His Gln Lys Lys Pro Tyr Cys His Ala His Asn Pro Lys50 55 60Asn Asn Thr Phe Thr Ser Val Tyr His Thr Pro Leu Asn Leu Asn65 70 75Val Arg Thr Phe Pro Glu Ala Ile Ser Gly Ile His Asp Gln Glu80 85 90Asp Gly Glu Gln Cys Lys Ser Val Phe His Trp Asp Met Lys Ser95 100 105Lys Asp Lys Glu Gly Ala Pro Asn Arg Gln Pro Leu Ala Asn Glu110 115 120Arg Ala Tyr Trp Thr Gly Tyr Gly Glu Gly Asn Ala Trp Cys Pro125 130 135
Gly Ala Leu Pro Asp Pro Glu Ile Val Arg Met Val Glu Ala Arg140 145 150Lys Ser Leu Gly Glu Glu Tyr Thr Glu Asp Tyr Glu Gln Pro Arg155 160 165Gly Lys Gly Ser Phe Pro Ala Met Ile Thr Pro Ala Tyr Gln Arg170 175 180Ala Lys Lys Ala Asn Gln Leu Ala Ser Gln Val Glu Tyr Lys Arg185 190 195Gly His Asp Glu Arg Ile Ser Arg Phe Ser Thr Val Ala Asp Thr200 205 210Pro Glu Leu Leu Arg Ser Lys Ala Gly Ala Gln Leu Gln Ser Asp215 220 225Val Arg Tyr Thr Glu Asp Tyr Glu Gln Gln Arg Gly Lys Gly Ser230 235 240Phe Pro Ala Met Ile Thr Pro Ala Tyr Gln Ile Ala Lys Arg Ala245 250 255Asn Glu Leu Ala Ser Asp Val Arg Tyr His Gln Gln Tyr Gln Lys260 265 270Glu Met Arg Gly Met Ala Gly Pro Ala Ile Gly Ala Glu Gly Ile275 280 285Leu Thr Arg Glu Cys Ala Asp Gln Tyr Gly Gln Gly Tyr Pro Glu290 295 300Glu Tyr Glu Glu His Arg Gly Lys Gly Ser Phe Pro Ala Met Ile305 310 315Thr Pro Ala Tyr Gln Asn Ala Lys Lys Ala His Glu Leu Ala Ser320 325 330Asp Ile Lys Tyr Arg Gln Asp Phe Asn Lys Met Lys Gly Thr Ala335 340 345His Tyr His Ser Leu Pro Ala Gln Asp Asn Leu Val Leu Lys Arg350 355 360Ala Gln Ser Val Asn Lys Leu Val Ser Glu Asn Lys Tyr Lys Glu365 370 375Asn Tyr Gln Asn His Met Arg Gly Arg Tyr Glu Gly Val Gly Met380 385 390Asp Arg Arg Thr Leu His Ala Met Lys Val Gly Ser Leu Ala Ser395 400 405Asn Val Ala Tyr Lys Ala Asp Tyr Lys His Asp Ile Val Asp Tyr410 415 420Asn Tyr Pro Ala Thr Leu Thr Pro Ser Tyr Gln Thr Ala Met Lys425 430 435Leu Val Pro Leu Lys Asp Ala Asn Tyr Arg Gln Ser Ile Asp Lys440 445 450Leu Lys Tyr Ser Ser Val Thr Asp Thr Pro Gln Ile Val Gln Ala455 460 465Lys Ile Asn Ala Gln Gln Leu Ser His Val Asn Tyr Arg Ala Asp470 475 480Tyr Glu Lys Asn Lys Leu Asn Tyr Thr Leu Pro Gln Asp Val Pro485 490 495
Gln Leu Val Lys Ala Lys Thr Asn Ala Lys Leu Phe Ser Glu Val500 505 510Lys Tyr Lys Glu Gly Trp Glu Lys Thr Lys Gly Lys Gly Phe Glu515 520 525Met Lys Leu Asp Ala Met Ser Leu Leu Ala Ala Lys Ala Ser Gly530 535 540Glu Leu Ala Ser Asn Ile Lys Tyr Lys Glu Glu Tyr Glu Lys Thr545 550 555Lys Gly Lys Ala Met Gly Thr Ala Asp Ser Arg Leu Leu His Ser560 565 570Leu Gln Ile Ala Lys Met Ser Ser Glu Val Glu Tyr Lys Lys Gly575 580 585Phe Glu Glu Ser Lys Thr Arg Phe His Leu Pro Met Asp Met Val590 595 600Asn Ile Arg His Ala Lys Lys Ala Gln Thr Leu Ala Ser Asp Leu605 610 615Asp Tyr Arg Lys Lys Leu His Glu Tyr Thr Val Leu Pro Glu Asp620 625 630Met Lys Thr Gln Trp Ala Lys Lys Val Tyr Gly Leu Gln Ser Glu635 640 645Leu Gln Tyr Lys Ala Asp Leu Ala Trp Met Lys Gly Val Gly Trp650 655 660Leu Thr Glu Gly Ser Leu Asn Leu Glu Gln Ala Lys Lys Ala Gly665 670 675Gln Leu Val Ser Glu Lys Asn Tyr Arg Gln Arg Val Asp Glu Leu680 685 690Lys Phe Thr Ser Val Thr Asp Ser Ser Gln Met Glu His Ala Lys695 700 705Lys Ser Gln Glu Leu Gln Ser Gly Val Ala Tyr Lys Ala Gly Asn710 715 720Glu Gln Ser Val His Gln Tyr Thr Ile Ser Lys Asp Glu Pro Leu725 730 735Phe Leu Gln Ala Arg Ala Asn Ala Ala Asn Leu Ser Glu Lys Leu740 745 750Tyr Lys Ser Ser Trp Glu Asn Gln Lys Ala Lys Gly Phe Glu Leu755 760 765Arg Leu Asp Ser Leu Thr Phe Leu Ala Ala Lys Ala Lys Arg Asp770 775 780Leu Ala Ser Glu Val Lys Tyr Lys Glu Asp Tyr Glu Arg Ser Arg785 790 795Gly Lys Leu Ile Gly Ala Lys Asp Val Gln Gly Asp Ser Gln Met800 805 810Ser His Ser Leu Gln Met Ser Lys Leu Gln Ser Glu Leu Glu Tyr815 820 825Lys Lys Gly Phe Glu Asp Thr Lys Ser Gln Cys His Val Ser Leu830 835 840Asp Met Val His Leu Val His Ala Arg Lys Ala Gln His Leu Ala845 850 855
Thr Asp Val Gly Tyr Lys Thr Ala Glu His His Phe Thr Ala Leu860 865 870Pro Thr Asp Met Lys Val Glu Trp Ala Lys Lys Ala Tyr Gly Leu875 880 885Gln Ser Asp Asn Gln Tyr Arg Ala Asp Val Lys Trp Met Lys Gly890 895 900Met Gly Trp Val Ala Thr Gly Ser Leu Asn Val Glu Gln Ala Lys905 910 915Lys Ala Gly Glu Leu Ile Ser Glu Lys Lys Tyr Arg Gln His Pro920 925 930Asp Ala Leu Lys Phe Thr Ser Ile Lys Asp Thr Pro Glu Met Val935 940 945Gln Ala Arg Ile Ser Tyr Thr Gln Ala Val Asp Arg Leu Tyr Arg950 955 960Glu Gln Gly Glu Asn Ile Lys His His Tyr Thr Pro Thr Ala Asp965 970 975Leu Pro Glu Val Leu Leu Ala Lys Leu Asn Ala Met Asn Ile Ser980 985 990Glu Thr Arg Tyr Lys Glu Ser Trp Ser Lys Leu Arg Asp Gly Gly99510001005Tyr Lys Leu Arg Leu Asp Ala Leu Pro Phe Gln Ala Ala Lys Ala101010151020Ser Gly Glu Ile Ile Ser Asp Tyr Lys Tyr Lys Glu Ala Phe Glu102510301035Lys Met Lys Gly Gln Met Leu Gly Ser Arg Ser Leu Glu Asp Asp104010451050Ile Ser Leu Ala His Ser Val Tyr Ala Thr Ser Leu Gln Ser Asp105510601065Val Asn Tyr Lys Lys Gly Phe Glu His Ser Lys Ala Gln Phe His107010751080Leu Pro Leu Asp Met Ala Ala Leu Val His Ala Lys Lys Ala Gln108510901095Thr Leu Ala Ser Asn Gln Asp Tyr Lys His Pro Leu Pro Gln Tyr110011051110Thr Ser Leu Ala Glu Asp Leu Arg Leu Ser Cys Ala Lys Lys Ala111511201125His Lys Leu Gln Ser Glu Asn Leu Tyr Arg Ser Asp Leu Asn Phe113011351140Met Arg Gly Val Ala Cys Val Ile Pro Gly Thr Leu Glu Ile Glu114511501155Gly Arg Lys Lys Ala Ser Glu Leu Ile Ser Glu Ser Lys Tyr Arg116011651170Gln His Pro His Ser Phe Lys Tyr Thr Ala Val Thr Asp Thr Pro117511801185Asn Leu Leu His Ala Lys Phe Ser Asn Gln Ile Thr Asn Glu Arg119011951200Leu Tyr Lys Ala Ala Gly Glu Asp Ala Arg His Glu Tyr Thr Met120512101215
Thr Leu Gly Leu Pro Glu Phe Ile Arg Ala Lys Thr Asn Ala Ala122012251230Asn Leu Ser Asp Ala Arg Tyr Lys Glu Ser Trp Arg Asn Leu Arg123512401245Ala Gln Gly Tyr Lys Leu Thr Ile Glu Ala Leu Pro Phe Gln Ala125012551260Ala Arg Ala Ser Gly Asp Ile Ala Ser Asp Phe Leu Tyr Arg His126512701275Asp Phe Val Lys Glu Arg Gly Lys Leu Ile Gly Pro Gln Ser Val128012851290Arg Asp Asp Pro Arg Ile Gln His Cys Arg Arg Met Gly Gln Leu129513001305Gln Ser Glu Leu Gln Tyr Arg Arg Gly Ala Thr Ser Ser Gln Ala131013151320Gln Phe His Leu Pro Met Asp Met Val His Leu Val His Ala Lys132513301335Asn Ala Gln Ala Leu Ala Ser Asp His Asp Tyr Arg Thr Gln Tyr134013451350His Lys Phe Thr Ala Leu Pro Glu Asp Leu Lys Met Ala Trp Ala135513601365Lys Lys Ala His Ala Leu Gln Ser Glu Leu Arg Tyr Lys Ser Asp137013751380Leu Ile Gly Met Lys Gly Ile Gly Trp Leu Ala Leu Arg Ser Pro138513901395Gln Met Glu Ser Ala Lys Lys Ala Gly Glu Leu Ile Ser Glu Thr140014051410Lys Tyr Arg Lys Lys Pro Asp Ser Ile Lys Phe Thr Thr Val Val141514201425Asp Ser Pro Asp Leu Val His Ala Lys Asn Ser Tyr Met His Cys143014351440Asn Glu Arg Met Tyr Arg Ser Gly Asp Ala Glu Ser Leu His Arg144514501455Tyr Thr Leu Ile Pro Asp His Pro Asp Phe Thr Arg Ala Arg Leu146014651470Asn Ala Leu His Leu Ser Asp Lys Val Tyr Arg Asn Ser Trp Glu147514801485Gln Thr Arg Ala Gly Ser Tyr Asp Phe Arg Leu Asp Ala Ile Pro149014951500Phe Gln Thr Ala Arg Ala Ser Arg Glu Ile Ala Ser Asp Phe Arg150515101515Tyr Lys Glu Ala Phe Leu Arg Asp Arg Gly Leu Gln Ile Gly Tyr152015251530Arg Ser Val Asp Asp Asp Pro Arg Met Lys His Phe Leu Asn Val153515401545Gly Arg Leu Gln Ser Asp Asn Glu Tyr Lys Lys Asp Phe Ala Lys155015551560Ser Arg Ser Gln Phe His Ser Ser Thr Asp Gln Pro Gly Leu Leu156515701575
Gln Ala Lys Arg Ser Gln Gln Leu Ala Ser Asp Val His Tyr Arg158015851590Gln Pro Leu Pro Gln Pro Thr Cys Asp Pro Glu Gln Leu Gly Leu159516001605Arg His Ala Gln Lys Ala His Gln Leu Gln Ser Asp Val Lys Tyr161016151620Lys Ser Asp Leu Asn Leu Thr Arg Gly Val Gly Trp Thr Pro Pro162516301635Gly Ser Tyr Lys Val Glu Met Ala Arg Arg Ala Ala Glu Leu Ala164016451650Asn Ala Arg Gly Leu Gly Leu Gln Gly Ala Tyr Arg Gly Ala Glu165516601665Ala Val Glu Ala Gly Asp His Gln Ser Gly Glu Val Asn Pro Asp167016751680Ala Thr Glu Ile Leu His Val Lys Lys Lys Lys Ala Leu Leu Leu16851690169权利要求
1.一种与骨骼肌和心肌功能相关的基因，它是下列核苷酸序列之一1)序列表中序列1的DNA序列；2)与序列表中序列1限定的DNA序列具有90％以上同源性，且编码相同功能蛋白质的DNA序列。
2.根据权利要求1所述的基因，其特征在于所述与骨骼肌和心肌功能相关的基因是序列表中序列1的DNA序列。
3.根据权利要求1或2所述的基因，其特征在于所述与骨骼肌和心肌功能相关基因的完整开放读码框为序列表中序列1的第166至5253共5088个碱基。
4.一种与骨骼肌和心肌功能相关的蛋白质，它具有序列表中序列2的氨基酸序列或将序列2的氨基酸序列经过一个或几个氨基酸残基的取代、缺失或添加且具有与序列2的氨基酸序列相同活性的由序列2衍生的蛋白质。
5.根据权利要求4所述的蛋白质，其特征在于所述蛋白质具有序列表中序列2的氨基酸序列。
全文摘要
本发明公开了一种骨骼肌和心肌功能相关基因及其编码的蛋白质。本发明涉及一种与骨骼肌和心肌功能相关的基因及其编码的蛋白质，特别是涉及一种与扩张型心肌病相关的基因及其编码的蛋白质。本发明的目的是提供一种与骨骼肌和心肌功能相关的基因及其编码的蛋白质。本发明所提供的与骨骼肌和心肌功能相关的基因是序列表中序列1的DNA序列；本发明所提供的蛋白质具有序列表中序列2的氨基酸序列。本发明的基因及其编码的蛋白质将为临床骨骼肌和心肌疾病的诊断和治疗提供有意义的生物学指标、药物靶标或蛋白类药物，并会带来巨大的社会效益及经济效益。
文档编号C12N15/12GK1536081SQ03109260
公开日2004年10月13日 申请日期2003年4月9日 优先权日2003年4月9日
发明者裴雪涛, 袁红丰, 李海民, 王冬梅 申请人:中国人民解放军军事医学科学院野战输血研究所, 中国人民解放军军事医学科学院野战输