CN113980971B

CN113980971B - 突变的马凡综合征致病基因fbn1及其应用

Info

Publication number: CN113980971B
Application number: CN202111286065.3A
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: Bestnovo Beijing Medical Technology Co Ltd
Current assignee: Bestnovo Beijing Medical Technology Co Ltd
Priority date: 2021-11-02
Filing date: 2021-11-02
Publication date: 2022-05-06
Anticipated expiration: 2041-11-02
Also published as: CN113980971A

Abstract

本发明涉及人类遗传学和内科心血管技术领域，具体涉及了一种突变的马凡综合征致病基因FBN1，与野生型FBN1基因编码DNA的参考序列相比，所述突变的马凡综合征致病基因FBN1的核苷酸序列为SEQ ID NO:3；在基因组位置chr15:48722984‑chr15:48722985处，缺失碱基TG；参考基因组版本是GRCh37。本发明还涉及上述突变的马凡综合征致病基因FBN1在制备马凡综合征检测试剂盒中的应用。本发明提供的突变的马凡综合征致病基因FBN1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，对马凡综合征的早期诊断，或者辅助临床判断具有重要意义。

Description

突变的马凡综合征致病基因FBN1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，尤其涉及突变的马凡综合征致病基因FBN1及其应用。

背景技术

遗传性主动脉疾病马凡综合征(Marfan syndrome，MFS)是一种遗传性结缔组织疾病，主要表现为骨骼、眼和心血管系统受累，易引发主动脉夹层和/或主动脉破裂，进而导致死亡。MFS患率0.065-0.2‰。此外，还有一些表型与MFS类似但患病率更低的其他综合征，如Loeys-Dietz综合征(LDS)、Shprintzen-Goldberg综合征(SGS)等。MFS的临床表型具有较大的异质性。

MFS主要为常染色体显性遗传，编码原纤维蛋白1的FBN1基因为其主要致病基因。根据《单基因遗传性心血管疾病基因诊断指南(2019)》，目前报道的FBN1基因相关突变超过1800个，符合临床诊断标准的MFS患者检出FBN1基因突变的比例为70-93％。

目前仍存在大量未知的FBN1基因突变位点，进一步发现新的FBN1基因的突变位点将有助于进一步研究马凡综合征，对马凡综合征的早期诊断，从分子水平揭示与FBN1基因相关的马凡综合征的发病机制，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马凡综合征，提供一种突变的马凡综合征致病基因FBN1及其应用。

本发明提供的技术方案如下：

一、本发明提供一种突变的马凡综合征致病基因FBN1，与野生型FBN1基因编码DNA的参考序列相比，所述突变的马凡综合征致病基因FBN1的核苷酸序列为SEQ ID NO:3；在基因组位置chr15:48722984-chr15:48722985处，缺失碱基TG；参考基因组版本是GRCh37。

二、本发明还提供了上述突变的马凡综合征致病基因FBN1在制备马凡综合征检测试剂盒中的应用。

优选地，所述马凡综合征检测试剂盒包括用于扩增致病基因FBN1的引物，引物的序列为SEQ ID NO:1和SEQ ID NO:2。

优选地，所述马凡综合征检测试剂盒还包括Taq DNA聚合酶和PCR缓冲液。

三、本发明的原理和有益效果在于：

本发明公开的突变的马凡综合征致病基因FBN1可以作为临床辅助诊断马凡综合征的生物标志物，对马凡综合征的早期诊断，或者辅助临床判断具有重要意义；基于突变的马凡综合征致病基因FBN1开发的检测试剂盒，可以检测出具有突变的马凡综合征致病基因FBN1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家系中先证者、先证者父亲等携带致病基因等人的Sanger测序图；

图3为实施例3家系中先证者母亲、先证者哥哥等人的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-突变的马凡综合征致病基因FBN1

突变的马凡综合征致病基因FBN1，具体突变如下表1所示：

表1突变的马凡综合征致病基因FBN1的具体检测结果

基因	基因组位置	转录本号	碱基改变	氨基酸改变	参考基因组版本
						FBN1	chr15:48722984-chr15:48722985	NM_000138	c.6753_6754del	p.Cys2251Ter	GRCh37/hg19

(1)在基因组位置chr15:48722967-chr15:48723016处，野生型FBN1基因的序列为：

为基因组位置chr15:48722984-chr15:48722985处突变前碱基；

在基因组位置chr15:48722967-chr15:48723014处，突变的马凡综合征致病基因FBN1的序列为：ttgtacattttttacagATGAGGATGAGTGAAGAGGGAAAACATGACT。

(2)野生型FBN1基因编码DNA的参考序列为：

为野生型FBN1基因编码DNA的参考序列的第6753和6754位突变前碱基。

c.6753_6754del表示：与野生型FBN1基因编码DNA的参考序列相比，突变的致病基因FBN1在第6753和6754位处缺失碱基TG，具体编码DNA的核苷酸序列为SEQ ID NO:3。

(3)野生型FBN1基因编码蛋白的氨基酸序列为：

为第2251位半胱氨酸(Cys,C)。

p.Cys2251Ter表示：第2251位的半胱氨酸(Cys,C)突变为终止密码子。突变FBN1基因编码蛋白的氨基酸序列为SEQ ID NO:4。

(4)受检者携带了突变FBN1基因c.6753_6754del杂合缺失变异(FBN1:p.Cys2251Ter het)，查询人群频率数据库发现该变异为罕见变异(千人基因组：无，ESP6500：无，ExAC：无)。该缺失变异使第2251位氨基酸处出现终止密码子，使蛋白截短表达。查询ClinVar、HGMD数据库未发现该变异，附近位点的其他无义变异c.6784C>T(p.Gln2262Ter)、c.6790G>T(p.Glu2264Ter)、c.6735C>A(p.Cys2245Ter)等被报导为马凡综合征的致病变异，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，使蛋白截短表达、附近位点多次被报导为致病变异，所以推断突变FBN1基因为马凡综合征的高度可疑致病突变。

实施例2-突变的马凡综合征致病基因FBN1在制备马凡综合征检测试剂盒中的应用

本实施例提供用于检测突变的致病基因FBN1的检测试剂盒，包括用于扩增致病基因FBN1的引物、Taq DNA聚合酶和PCR缓冲液等。

引物具体如下：

上游引物(FBN1-E56F，SEQ ID NO:1)：5'AGCCAGTAGTGAAATAACAGA 3'；

下游引物(FBN1-E56R，SEQ ID NO:2)：5'TCGCCAAGAACAGTATCCC 3'；

长度：353bp。

利用本试剂盒的具体检测步骤为：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FBN1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FBN1基因是否携带c.6753_6754del杂合错义变异，协助临床确诊待测者是否患有具有c.6753_6754del FBN1基因突变的患者。

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突变的致病基因FBN1的致病性。

具体地，选取一个马凡综合征家系中的三代成员，该家系中的先证者在中国医学科学院阜外医院治疗，临床诊断患有马凡综合征。

样本来源：阜外医院，在先证者(女，4岁)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。

表2先证者病史简述

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FBN1基因检测，检测结果如图1-图3所示。图1为实施例3的家系图；图2为实施例3家系中先证者、先证者父亲等携带致病基因等人的Sanger测序图；图3为实施例3家系中先证者母亲、先证者哥哥等人的Sanger测序图。

如图1-图3所示，该家系中临床诊断出马凡综合征的先证者、先证者父亲等人均携带了突变的致病基因FBN1，而未患有马凡综合征的先证者母亲、先证者哥哥等人均为携带突变的致病基因FBN1。

该家系验证说明了突变FBN1致病基因对马凡综合征的致病性，支持临床诊断。

实施例4-针对家系外正常人的验证实验

采用实施例2的突变的马凡综合征致病基因FBN1体外检测试剂盒，对1500例家系外正常人进行FBN1基因检测，结果均未能检测到该突变。

实施例5-针对家系外马凡综合征患者的验证实验

在中国范围内，对患有马凡综合征、肥厚型心肌病、扩张型心肌病、长QT等遗传病的患者中检测FBN1基因，患者总共2500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，仅在临床诊断患有马凡综合征的3例患者中检测到具有c.6753_6754del杂合错义的突变的致病基因FBN1。

本实验再次验证说明突变的FBN1基因的c.6753_6754del杂合错义变异会导致马凡综合征，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。

序列表

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cgggccaaga gaagaggcgg tggaggacac gacgcgctta aaggacccaa tgtctgtgga 180

tcacgttata atgcttactg ttgccctgga tggaaaacct tacctggcgg aaatcagtgt 240

attgtcccca tttgccggca ttcctgtggg gatggatttt gttcgaggcc aaatatgtgc 300

acttgcccat ctggtcagat agctccttcc tgtggctcca gatccataca acactgcaat 360

attcgctgta tgaatggagg tagctgcagt gacgatcact gtctatgcca gaaaggatac 420

atagggactc actgtggaca acctgtttgt gaaagtggct gtctcaatgg aggaaggtgt 480

gtggccccaa atcgatgtgc atgcacttac ggatttactg gaccccagtg tgaaagagat 540

tacaggacag gcccatgttt tactgtgatc agcaaccaga tgtgccaggg acaactcagc 600

gggattgtct gcacaaaaac gctctgctgt gccacagtcg gccgagcctg gggccacccc 660

tgtgagatgt gtcctgccca gcctcacccc tgccgccgtg gcttcattcc aaatatccgc 720

acgggagctt gtcaagatgt ggatgaatgc caggccatcc ccgggctctg tcagggagga 780

aattgcatta atactgttgg gtcttttgag tgcaaatgcc ctgctggaca caaacttaat 840

gaagtgtcac aaaaatgtga agatattgat gaatgcagca ccattcctgg aatctgtgaa 900

gggggtgaat gtacaaacac agtcagcagt tacttttgca aatgtccccc tggtttttac 960

acctctccag atggtaccag atgcatagat gttcgcccag gatactgtta cacagctctg 1020

acaaacgggc gctgctctaa ccagctgcca cagtccataa ccaaaatgca gtgctgctgt 1080

gatgccggcc gatgctggtc tccaggggtc actgtcgccc ctgagatgtg tcccatcaga 1140

gcaaccgagg atttcaacaa gctgtgctct gttcctatgg taattcctgg gagaccagaa 1200

tatcctcccc caccccttgg ccccattcct ccagttctcc ctgttcctcc tggctttcct 1260

cctggacctc aaattccggt ccctcgacca ccagtggaat atctgtatcc atctcgggag 1320

ccaccaaggg tgctgccagt aaacgttact gattactgcc agttggtccg ctatctctgt 1380

caaaatggac gctgcattcc aactcctggg agttaccggt gtgagtgcaa caaagggttc 1440

cagctggacc tccgtgggga gtgtattgat gttgatgaat gtgagaaaaa cccctgtgct 1500

ggtggtgagt gtattaacaa ccagggttcg tacacctgtc agtgccgagc tggatatcag 1560

agcacactca cgcggacaga atgccgagac attgatgagt gtttacagaa tggccggatc 1620

tgcaataatg gacgctgcat caacacagat ggcagttttc attgcgtgtg taatgcgggc 1680

tttcatgtta cacgagatgg gaagaactgt gaagatatgg atgaatgcag cataaggaac 1740

atgtgcctta atggaatgtg tatcaatgaa gatggcagtt ttaaatgtat ttgcaaacct 1800

ggattccagc tggcatcaga tggacgttat tgcaaagaca ttaacgagtg tgaaacccct 1860

gggatctgca tgaatgggcg ttgcgtcaac actgatggct cctacagatg tgaatgcttc 1920

cctggactgg ctgtgggtct ggatggccgt gtgtgtgttg acacacacat gcggagcaca 1980

tgctatggtg gatacaagag aggccagtgt atcaaacctt tgtttggtgc tgtcactaaa 2040

tctgaatgct gttgcgccag cactgagtat gcatttgggg aaccttgcca gccgtgtcct 2100

gcacagaatt cagcggaata tcaggcactc tgcagcagtg ggccaggaat gacgtcagca 2160

ggcagtgata taaatgaatg tgcactagat cctgatattt gcccaaatgg aatctgtgaa 2220

aaccttcgtg ggacctataa atgtatatgc aattcaggat atgaagtgga ttcaactggg 2280

aaaaactgcg ttgatattaa tgaatgtgta ctgaacagtc tcctttgtga caatggacaa 2340

tgtagaaata ctcctggaag ttttgtctgt acctgcccca agggatttat ctacaaacct 2400

gatctaaaaa catgtgaaga cattgatgaa tgcgaatcaa gtccttgcat taatggagtc 2460

tgcaagaaca gcccaggctc ttttatttgt gaatgttctt ctgaaagtac tttggatcca 2520

acaaaaacca tctgcataga aaccatcaag ggcacttgct ggcagactgt cattgatggg 2580

cgatgtgaga tcaacatcaa tggagccacc ttaaagtccc agtgctgctc ctccctcggt 2640

gctgcgtggg gaagcccgtg caccctatgc caagttgatc ccatatgtgg taaagggtac 2700

tcaagaatta aaggaacaca atgtgaagat atagatgaat gtgaagtgtt cccaggagtg 2760

tgtaaaaatg gcctgtgtgt taacactagg gggtcattca agtgtcagtg tcccagtgga 2820

atgactttgg atgccacagg aaggatctgt cttgatatcc gcctggaaac ctgcttcctg 2880

aggtacgagg acgaggagtg caccctgcct attgctggcc gccaccgcat ggacgcctgc 2940

tgctgctccg tcggggcagc ctggggtact gaggaatgcg aggagtgtcc catgagaaat 3000

actcctgagt acgaggagct gtgtccgaga ggacccggat ttgccacaaa agaaattaca 3060

aatggaaagc ctttcttcaa agatatcaat gagtgcaaga tgatacccag cctctgcacc 3120

cacggcaagt gcagaaacac cattggcagc tttaagtgca ggtgtgacag cggctttgct 3180

cttgattctg aagaaaggaa ctgcacagac attgacgaat gccgcatatc tcctgacctc 3240

tgtggcagag gccagtgtgt gaacacccct ggggactttg aatgcaagtg tgacgaaggc 3300

tatgaaagtg gattcatgat gatgaagaac tgcatggata ttgatgagtg tcagagagat 3360

cctctcctat gccgaggtgg tgtttgccat aacacagagg gaagttaccg ctgtgaatgc 3420

ccgcctggcc atcagctgtc ccccaacatc tccgcgtgta tcgacatcaa tgaatgtgag 3480

ctgagtgcac acctgtgccc caatggccgt tgcgtgaacc tcatagggaa gtatcagtgt 3540

gcctgcaacc ctggctacca ttcaactccc gataggctat tttgtgttga cattgatgaa 3600

tgcagcataa tgaatggtgg ttgtgaaacc ttctgcacaa actctgaagg cagctatgaa 3660

tgtagctgtc agccgggatt tgcactaatg cctgaccaga gatcatgcac cgacatcgat 3720

gagtgtgaag ataatcccaa tatctgtgat ggtggtcagt gcacaaatat ccctggagag 3780

tacaggtgct tgtgttatga tggattcatg gcatctgaag acatgaagac ttgtgtagat 3840

gtcaatgagt gtgacctgaa tccaaatatc tgcctaagtg ggacctgtga aaacacgaaa 3900

ggctcattta tctgccactg tgatatgggc tactccggca aaaaaggaaa aactggctgt 3960

acagacatca atgaatgtga aattggagca cacaactgtg gcaaacatgc tgtatgtacc 4020

aatacagcag gaagcttcaa atgtagctgc agtcccgggt ggattggaga tggcattaag 4080

tgcactgatc tggacgaatg ttccaatgga acccatatgt gcagccagca tgcagactgc 4140

aagaatacca tgggatctta ccgctgtctg tgcaaggaag gatacacagg tgatggcttc 4200

acttgtacag accttgatga gtgctctgag aacctgaatc tctgtggcaa tggccagtgc 4260

ctcaatgcac caggaggata ccgctgtgaa tgcgacatgg gcttcgtgcc cagtgctgac 4320

gggaaagcct gtgaagatat tgatgagtgc tcccttccga acatctgtgt ctttggaact 4380

tgccacaacc tccctggcct gttccgctgt gagtgtgaga taggctacga actggacaga 4440

agcggcggga actgcacaga tgtgaatgaa tgcctggatc caaccacgtg catcagtggg 4500

aactgtgtca acactccagg cagctatatc tgtgactgcc cacctgattt tgaactgaac 4560

ccaactcgag ttggctgtgt tgatacccgc tctggaaatt gctatttgga tattcgacct 4620

cgaggagaca atggagatac agcctgcagc aatgaaattg gagttggtgt ttccaaagct 4680

tcctgctgct gttctctggg taaagcctgg ggtactcctt gtgagatgtg tcctgctgtg 4740

aacacatccg agtacaaaat tctttgtcct ggaggggaag gtttccgacc aaatcctatc 4800

accgttatat tggaagatat tgatgagtgc caggagctac cagggctgtg ccaaggagga 4860

aaatgtatca acacctttgg gagtttccag tgccgctgtc caaccggcta ctacctgaat 4920

gaagatacac gagtgtgtga tgatgtgaat gaatgtgaga ctcctggaat ctgtggtcca 4980

gggacatgtt acaacaccgt tggcaactac acctgtatct gtcctccaga ctacatgcaa 5040

gtgaatgggg gaaataattg catggatatg agaagaagtt tgtgctacag aaactactat 5100

gctgacaacc agacctgtga tggagaattg ttattcaaca tgaccaagaa gatgtgctgc 5160

tgttcctaca acattggccg ggcgtggaac aagccctgtg aacagtgtcc catcccaagt 5220

acagatgagt ttgctacact ctgtggaagt caaaggccag gctttgtcat cgacatttat 5280

accggtttac ccgttgatat tgatgagtgc cgggagatcc caggggtctg tgaaaatgga 5340

gtgtgtatca acatggttgg cagcttccga tgtgaatgtc cagtgggatt cttctataat 5400

gacaagttgt tggtttgtga agatattgac gagtgtcaga acggcccagt gtgccagcgc 5460

aacgccgaat gcatcaacac tgcaggcagc taccgctgtg actgtaagcc cggctaccgc 5520

ttcacctcca caggacagtg caatgatcgt aatgaatgtc aagaaatccc caatatatgc 5580

agtcatgggc agtgcattga cacagttgga agcttttatt gcctttgcca cactggtttt 5640

aaaacaaatg atgaccaaac catgtgcttg gacataaatg aatgtgaaag agatgcctgt 5700

gggaatggaa cttgccggaa cacaattggt tccttcaact gccgctgcaa tcatggtttc 5760

atcctttctc acaacaatga ctgtatagat gttgatgaat gtgcaagtgg aaatgggaat 5820

ctttgcagaa atggccaatg cattaataca gtggggtctt tccagtgcca gtgcaatgaa 5880

ggctatgagg tggctccaga tgggaggacc tgtgtggata tcaatgaatg tcttctagaa 5940

cccagaaaat gtgcaccagg tacctgtcaa aacttggatg ggtcctacag atgcatttgc 6000

ccacctggat acagtcttca aaatgagaag tgtgaagata ttgatgagtg tgtcgaagag 6060

ccagaaattt gtgccctggg cacatgcagt aacactgaag gcagcttcaa atgtctgtgt 6120

ccagaagggt tttccttgtc ctccagtgga agaaggtgcc aagatttgcg aatgagctac 6180

tgttatgcga agtttgaagg aggaaagtgt tcatcaccca aatccagaaa tcactccaag 6240

caggaatgct gctgtgcctt gaagggagaa ggctggggag acccctgcga gctctgcccc 6300

acggaacctg atgaggcctt ccgccagata tgtccttatg gaagtgggat catcgtggga 6360

cctgatgatt cagcagttga tatggacgaa tgcaaagaac ccgatgtctg taaacatgga 6420

cagtgcatca atacagatgg ttcctatcgc tgcgagtgtc cctttggtta tattctagca 6480

gggaatgaat gtgtagatac tgatgaatgt tctgttggca atccttgtgg aaatggaacc 6540

tgcaagaatg tgattggagg ttttgaatgc acctgcgagg agggatttga gcccggtcca 6600

atgatgacat gtgaagatat aaatgaatgt gcccagaatc ctctgctctg tgccttccga 6660

tgtgtgaaca cttatgggtc atatgaatgc aaatgtcccg tgggatatgt gctcagagaa 6720

gaccgtagga tgtgcaaaga tgaggatgag tga 6753

<210> 4

<211> 2250

<212> PRT

<213> homo sapiens

<400> 4

Met Arg Arg Gly Arg Leu Leu Glu Ile Ala Leu Gly Phe Thr Val Leu

1 5 10 15

Leu Ala Ser Tyr Thr Ser His Gly Ala Asp Ala Asn Leu Glu Ala Gly

20 25 30

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

35 40 45

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

50 55 60

Ala Tyr Cys Cys Pro Gly Trp Lys Thr Leu Pro Gly Gly Asn Gln Cys

65 70 75 80

Ile Val Pro Ile Cys Arg His Ser Cys Gly Asp Gly Phe Cys Ser Arg

85 90 95

Pro Asn Met Cys Thr Cys Pro Ser Gly Gln Ile Ala Pro Ser Cys Gly

100 105 110

Ser Arg Ser Ile Gln His Cys Asn Ile Arg Cys Met Asn Gly Gly Ser

115 120 125

Cys Ser Asp Asp His Cys Leu Cys Gln Lys Gly Tyr Ile Gly Thr His

130 135 140

Cys Gly Gln Pro Val Cys Glu Ser Gly Cys Leu Asn Gly Gly Arg Cys

145 150 155 160

Val Ala Pro Asn Arg Cys Ala Cys Thr Tyr Gly Phe Thr Gly Pro Gln

165 170 175

Cys Glu Arg Asp Tyr Arg Thr Gly Pro Cys Phe Thr Val Ile Ser Asn

180 185 190

Gln Met Cys Gln Gly Gln Leu Ser Gly Ile Val Cys Thr Lys Thr Leu

195 200 205

Cys Cys Ala Thr Val Gly Arg Ala Trp Gly His Pro Cys Glu Met Cys

210 215 220

Pro Ala Gln Pro His Pro Cys Arg Arg Gly Phe Ile Pro Asn Ile Arg

225 230 235 240

Thr Gly Ala Cys Gln Asp Val Asp Glu Cys Gln Ala Ile Pro Gly Leu

245 250 255

Cys Gln Gly Gly Asn Cys Ile Asn Thr Val Gly Ser Phe Glu Cys Lys

260 265 270

Cys Pro Ala Gly His Lys Leu Asn Glu Val Ser Gln Lys Cys Glu Asp

275 280 285

Ile Asp Glu Cys Ser Thr Ile Pro Gly Ile Cys Glu Gly Gly Glu Cys

290 295 300

Thr Asn Thr Val Ser Ser Tyr Phe Cys Lys Cys Pro Pro Gly Phe Tyr

305 310 315 320

Thr Ser Pro Asp Gly Thr Arg Cys Ile Asp Val Arg Pro Gly Tyr Cys

325 330 335

Tyr Thr Ala Leu Thr Asn Gly Arg Cys Ser Asn Gln Leu Pro Gln Ser

340 345 350

Ile Thr Lys Met Gln Cys Cys Cys Asp Ala Gly Arg Cys Trp Ser Pro

355 360 365

Gly Val Thr Val Ala Pro Glu Met Cys Pro Ile Arg Ala Thr Glu Asp

370 375 380

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

385 390 395 400

Tyr Pro Pro Pro Pro Leu Gly Pro Ile Pro Pro Val Leu Pro Val Pro

405 410 415

Pro Gly Phe Pro Pro Gly Pro Gln Ile Pro Val Pro Arg Pro Pro Val

420 425 430

Glu Tyr Leu Tyr Pro Ser Arg Glu Pro Pro Arg Val Leu Pro Val Asn

435 440 445

Val Thr Asp Tyr Cys Gln Leu Val Arg Tyr Leu Cys Gln Asn Gly Arg

450 455 460

Cys Ile Pro Thr Pro Gly Ser Tyr Arg Cys Glu Cys Asn Lys Gly Phe

465 470 475 480

Gln Leu Asp Leu Arg Gly Glu Cys Ile Asp Val Asp Glu Cys Glu Lys

485 490 495

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

500 505 510

Cys Gln Cys Arg Ala Gly Tyr Gln Ser Thr Leu Thr Arg Thr Glu Cys

515 520 525

Arg Asp Ile Asp Glu Cys Leu Gln Asn Gly Arg Ile Cys Asn Asn Gly

530 535 540

Arg Cys Ile Asn Thr Asp Gly Ser Phe His Cys Val Cys Asn Ala Gly

545 550 555 560

Phe His Val Thr Arg Asp Gly Lys Asn Cys Glu Asp Met Asp Glu Cys

565 570 575

Ser Ile Arg Asn Met Cys Leu Asn Gly Met Cys Ile Asn Glu Asp Gly

580 585 590

Ser Phe Lys Cys Ile Cys Lys Pro Gly Phe Gln Leu Ala Ser Asp Gly

595 600 605

Arg Tyr Cys Lys Asp Ile Asn Glu Cys Glu Thr Pro Gly Ile Cys Met

610 615 620

Asn Gly Arg Cys Val Asn Thr Asp Gly Ser Tyr Arg Cys Glu Cys Phe

625 630 635 640

Pro Gly Leu Ala Val Gly Leu Asp Gly Arg Val Cys Val Asp Thr His

645 650 655

Met Arg Ser Thr Cys Tyr Gly Gly Tyr Lys Arg Gly Gln Cys Ile Lys

660 665 670

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

675 680 685

Glu Tyr Ala Phe Gly Glu Pro Cys Gln Pro Cys Pro Ala Gln Asn Ser

690 695 700

Ala Glu Tyr Gln Ala Leu Cys Ser Ser Gly Pro Gly Met Thr Ser Ala

705 710 715 720

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

725 730 735

Gly Ile Cys Glu Asn Leu Arg Gly Thr Tyr Lys Cys Ile Cys Asn Ser

740 745 750

Gly Tyr Glu Val Asp Ser Thr Gly Lys Asn Cys Val Asp Ile Asn Glu

755 760 765

Cys Val Leu Asn Ser Leu Leu Cys Asp Asn Gly Gln Cys Arg Asn Thr

770 775 780

Pro Gly Ser Phe Val Cys Thr Cys Pro Lys Gly Phe Ile Tyr Lys Pro

785 790 795 800

Asp Leu Lys Thr Cys Glu Asp Ile Asp Glu Cys Glu Ser Ser Pro Cys

805 810 815

Ile Asn Gly Val Cys Lys Asn Ser Pro Gly Ser Phe Ile Cys Glu Cys

820 825 830

Ser Ser Glu Ser Thr Leu Asp Pro Thr Lys Thr Ile Cys Ile Glu Thr

835 840 845

Ile Lys Gly Thr Cys Trp Gln Thr Val Ile Asp Gly Arg Cys Glu Ile

850 855 860

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

865 870 875 880

Ala Ala Trp Gly Ser Pro Cys Thr Leu Cys Gln Val Asp Pro Ile Cys

885 890 895

Gly Lys Gly Tyr Ser Arg Ile Lys Gly Thr Gln Cys Glu Asp Ile Asp

900 905 910

Glu Cys Glu Val Phe Pro Gly Val Cys Lys Asn Gly Leu Cys Val Asn

915 920 925

Thr Arg Gly Ser Phe Lys Cys Gln Cys Pro Ser Gly Met Thr Leu Asp

930 935 940

Ala Thr Gly Arg Ile Cys Leu Asp Ile Arg Leu Glu Thr Cys Phe Leu

945 950 955 960

Arg Tyr Glu Asp Glu Glu Cys Thr Leu Pro Ile Ala Gly Arg His Arg

965 970 975

Met Asp Ala Cys Cys Cys Ser Val Gly Ala Ala Trp Gly Thr Glu Glu

980 985 990

Cys Glu Glu Cys Pro Met Arg Asn Thr Pro Glu Tyr Glu Glu Leu Cys

995 1000 1005

Pro Arg Gly Pro Gly Phe Ala Thr Lys Glu Ile Thr Asn Gly Lys Pro

1010 1015 1020

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

1025 1030 1035 1040

His Gly Lys Cys Arg Asn Thr Ile Gly Ser Phe Lys Cys Arg Cys Asp

1045 1050 1055

Ser Gly Phe Ala Leu Asp Ser Glu Glu Arg Asn Cys Thr Asp Ile Asp

1060 1065 1070

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

1075 1080 1085

Thr Pro Gly Asp Phe Glu Cys Lys Cys Asp Glu Gly Tyr Glu Ser Gly

1090 1095 1100

Phe Met Met Met Lys Asn Cys Met Asp Ile Asp Glu Cys Gln Arg Asp

1105 1110 1115 1120

Pro Leu Leu Cys Arg Gly Gly Val Cys His Asn Thr Glu Gly Ser Tyr

1125 1130 1135

Arg Cys Glu Cys Pro Pro Gly His Gln Leu Ser Pro Asn Ile Ser Ala

1140 1145 1150

Cys Ile Asp Ile Asn Glu Cys Glu Leu Ser Ala His Leu Cys Pro Asn

1155 1160 1165

Gly Arg Cys Val Asn Leu Ile Gly Lys Tyr Gln Cys Ala Cys Asn Pro

1170 1175 1180

Gly Tyr His Ser Thr Pro Asp Arg Leu Phe Cys Val Asp Ile Asp Glu

1185 1190 1195 1200

Cys Ser Ile Met Asn Gly Gly Cys Glu Thr Phe Cys Thr Asn Ser Glu

1205 1210 1215

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

1220 1225 1230

Gln Arg Ser Cys Thr Asp Ile Asp Glu Cys Glu Asp Asn Pro Asn Ile

1235 1240 1245

Cys Asp Gly Gly Gln Cys Thr Asn Ile Pro Gly Glu Tyr Arg Cys Leu

1250 1255 1260

Cys Tyr Asp Gly Phe Met Ala Ser Glu Asp Met Lys Thr Cys Val Asp

1265 1270 1275 1280

Val Asn Glu Cys Asp Leu Asn Pro Asn Ile Cys Leu Ser Gly Thr Cys

1285 1290 1295

Glu Asn Thr Lys Gly Ser Phe Ile Cys His Cys Asp Met Gly Tyr Ser

1300 1305 1310

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

1315 1320 1325

Gly Ala His Asn Cys Gly Lys His Ala Val Cys Thr Asn Thr Ala Gly

1330 1335 1340

Ser Phe Lys Cys Ser Cys Ser Pro Gly Trp Ile Gly Asp Gly Ile Lys

1345 1350 1355 1360

Cys Thr Asp Leu Asp Glu Cys Ser Asn Gly Thr His Met Cys Ser Gln

1365 1370 1375

His Ala Asp Cys Lys Asn Thr Met Gly Ser Tyr Arg Cys Leu Cys Lys

1380 1385 1390

Glu Gly Tyr Thr Gly Asp Gly Phe Thr Cys Thr Asp Leu Asp Glu Cys

1395 1400 1405

Ser Glu Asn Leu Asn Leu Cys Gly Asn Gly Gln Cys Leu Asn Ala Pro

1410 1415 1420

Gly Gly Tyr Arg Cys Glu Cys Asp Met Gly Phe Val Pro Ser Ala Asp

1425 1430 1435 1440

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

1445 1450 1455

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

1460 1465 1470

Glu Ile Gly Tyr Glu Leu Asp Arg Ser Gly Gly Asn Cys Thr Asp Val

1475 1480 1485

Asn Glu Cys Leu Asp Pro Thr Thr Cys Ile Ser Gly Asn Cys Val Asn

1490 1495 1500

Thr Pro Gly Ser Tyr Ile Cys Asp Cys Pro Pro Asp Phe Glu Leu Asn

1505 1510 1515 1520

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

1525 1530 1535

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

1540 1545 1550

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

1555 1560 1565

Ala Trp Gly Thr Pro Cys Glu Met Cys Pro Ala Val Asn Thr Ser Glu

1570 1575 1580

Tyr Lys Ile Leu Cys Pro Gly Gly Glu Gly Phe Arg Pro Asn Pro Ile

1585 1590 1595 1600

Thr Val Ile Leu Glu Asp Ile Asp Glu Cys Gln Glu Leu Pro Gly Leu

1605 1610 1615

Cys Gln Gly Gly Lys Cys Ile Asn Thr Phe Gly Ser Phe Gln Cys Arg

1620 1625 1630

Cys Pro Thr Gly Tyr Tyr Leu Asn Glu Asp Thr Arg Val Cys Asp Asp

1635 1640 1645

Val Asn Glu Cys Glu Thr Pro Gly Ile Cys Gly Pro Gly Thr Cys Tyr

1650 1655 1660

Asn Thr Val Gly Asn Tyr Thr Cys Ile Cys Pro Pro Asp Tyr Met Gln

1665 1670 1675 1680

Val Asn Gly Gly Asn Asn Cys Met Asp Met Arg Arg Ser Leu Cys Tyr

1685 1690 1695

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

1700 1705 1710

Asn Met Thr Lys Lys Met Cys Cys Cys Ser Tyr Asn Ile Gly Arg Ala

1715 1720 1725

Trp Asn Lys Pro Cys Glu Gln Cys Pro Ile Pro Ser Thr Asp Glu Phe

1730 1735 1740

Ala Thr Leu Cys Gly Ser Gln Arg Pro Gly Phe Val Ile Asp Ile Tyr

1745 1750 1755 1760

Thr Gly Leu Pro Val Asp Ile Asp Glu Cys Arg Glu Ile Pro Gly Val

1765 1770 1775

Cys Glu Asn Gly Val Cys Ile Asn Met Val Gly Ser Phe Arg Cys Glu

1780 1785 1790

Cys Pro Val Gly Phe Phe Tyr Asn Asp Lys Leu Leu Val Cys Glu Asp

1795 1800 1805

Ile Asp Glu Cys Gln Asn Gly Pro Val Cys Gln Arg Asn Ala Glu Cys

1810 1815 1820

Ile Asn Thr Ala Gly Ser Tyr Arg Cys Asp Cys Lys Pro Gly Tyr Arg

1825 1830 1835 1840

Phe Thr Ser Thr Gly Gln Cys Asn Asp Arg Asn Glu Cys Gln Glu Ile

1845 1850 1855

Pro Asn Ile Cys Ser His Gly Gln Cys Ile Asp Thr Val Gly Ser Phe

1860 1865 1870

Tyr Cys Leu Cys His Thr Gly Phe Lys Thr Asn Asp Asp Gln Thr Met

1875 1880 1885

Cys Leu Asp Ile Asn Glu Cys Glu Arg Asp Ala Cys Gly Asn Gly Thr

1890 1895 1900

Cys Arg Asn Thr Ile Gly Ser Phe Asn Cys Arg Cys Asn His Gly Phe

1905 1910 1915 1920

Ile Leu Ser His Asn Asn Asp Cys Ile Asp Val Asp Glu Cys Ala Ser

1925 1930 1935

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

1940 1945 1950

Ser Phe Gln Cys Gln Cys Asn Glu Gly Tyr Glu Val Ala Pro Asp Gly

1955 1960 1965

Arg Thr Cys Val Asp Ile Asn Glu Cys Leu Leu Glu Pro Arg Lys Cys

1970 1975 1980

Ala Pro Gly Thr Cys Gln Asn Leu Asp Gly Ser Tyr Arg Cys Ile Cys

1985 1990 1995 2000

Pro Pro Gly Tyr Ser Leu Gln Asn Glu Lys Cys Glu Asp Ile Asp Glu

2005 2010 2015

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

2020 2025 2030

Glu Gly Ser Phe Lys Cys Leu Cys Pro Glu Gly Phe Ser Leu Ser Ser

2035 2040 2045

Ser Gly Arg Arg Cys Gln Asp Leu Arg Met Ser Tyr Cys Tyr Ala Lys

2050 2055 2060

Phe Glu Gly Gly Lys Cys Ser Ser Pro Lys Ser Arg Asn His Ser Lys

2065 2070 2075 2080

Gln Glu Cys Cys Cys Ala Leu Lys Gly Glu Gly Trp Gly Asp Pro Cys

2085 2090 2095

Glu Leu Cys Pro Thr Glu Pro Asp Glu Ala Phe Arg Gln Ile Cys Pro

2100 2105 2110

Tyr Gly Ser Gly Ile Ile Val Gly Pro Asp Asp Ser Ala Val Asp Met

2115 2120 2125

Asp Glu Cys Lys Glu Pro Asp Val Cys Lys His Gly Gln Cys Ile Asn

2130 2135 2140

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

2145 2150 2155 2160

Gly Asn Glu Cys Val Asp Thr Asp Glu Cys Ser Val Gly Asn Pro Cys

2165 2170 2175

Gly Asn Gly Thr Cys Lys Asn Val Ile Gly Gly Phe Glu Cys Thr Cys

2180 2185 2190

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

2195 2200 2205

Glu Cys Ala Gln Asn Pro Leu Leu Cys Ala Phe Arg Cys Val Asn Thr

2210 2215 2220

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

2225 2230 2235 2240

Asp Arg Arg Met Cys Lys Asp Glu Asp Glu

2245 2250

Claims

1.突变的马凡综合征致病基因FBN1，其特征在于，与野生型FBN1基因编码DNA的参考序列相比，突变的致病基因FBN1在第6753和6754位处缺失碱基TG，编码DNA的核苷酸序列为SEQ ID NO:3。