CN114317466B

CN114317466B - 一种突变acc1蛋白及其用途

Info

Publication number: CN114317466B
Application number: CN202210002987.5A
Authority: CN
Inventors: 徐芬; 冼颖欣; 曹欢易; 梁华; 蔡梦茵
Original assignee: Third Affiliated Hospital Sun Yat Sen University
Current assignee: Third Affiliated Hospital Sun Yat Sen University
Priority date: 2022-01-04
Filing date: 2022-01-04
Publication date: 2023-10-27
Anticipated expiration: 2042-01-04
Also published as: CN114317466A

Abstract

本发明涉及医学检测领域，尤其涉及一种突变ACC1蛋白及其用途。本发明的突变ACC1蛋白为将野生型ACC1蛋白的第1523位赖氨酸突变为谷氨酸得到，所述野生型ACC1蛋白的氨基酸序列如SEQ ID NO：1所示。利用蛋白质组学和生信分析发现并鉴定NAFLD小鼠模型的肝脏中发生显著差异丙二酰化修饰的ACC1蛋白及其丙二酰化位点K1523，说明ACC1蛋白丙二酰化修饰与肝脏脂质沉积有关，将来可作为协助诊断和判断病情严重程度的生物学标记物。利用K1523E突变模拟ACC1蛋白丙二酰化修饰，证明K1523是ACC1发挥酶活性的关键位点，将来可作为抗体制备、NAFLD治疗药物研发的靶点。

Description

一种突变ACC1蛋白及其用途

技术领域

本发明涉及医学检测领域，尤其涉及一种突变ACC1蛋白及其用途。

背景技术

非酒精性脂肪性肝病(NAFLD)以肝脏脂质异常沉积为主要特征，疾病谱包括单纯性脂肪肝、非酒精性脂肪性肝炎(NASH)以及脂肪性肝硬化。其中肝脏脂质从头合成过程增强是导致脂质沉积的最主要原因。在胰岛素抵抗状态下，胰岛素抑制脂肪酸合成的作用受损，肝脏脂质从头合成过程增强，使得甘油三酯在肝脏异常积累、超过肝脏处理脂肪酸的能力，导致肝脏脂肪变性。因此，胰岛素抵抗是NAFLD的发病机制中关键的一环。

NAFLD的诊断金标准为肝脏穿刺活检，但NAFLD常常为隐匿起病，无特异性症状和体征，肝脏穿刺活检为高侵入性的诊断方法，存在疼痛及其他风险，患者常常不愿意接受。此外，NAFLD患者的肝转氨酶ALT、GGT常有升高，但缺乏特异性。因此，NAFLD需要一种方便且具有特异性的诊断方法，对NAFLD的防治具有重要意义。NAFLD的治疗仍以生活、饮食干预为主，至今尚缺乏FDA正式批准用于治疗NAFLD的药物，生活方式和饮食干预仍然是最有效、安全的治疗方法。

ACC1(acetyl-CoA carboxylase 1，乙酰辅酶A羧化酶1)是脂肪酸生物合成的限速酶，是碳水化合物转化为脂肪酸的重要调控分子。ACC1能催化乙酰辅酶A羧化反应生成丙二酰辅酶A，后者成为肝脏脂质从头合成的直接供体，因此，ACC1在NAFLD发病过程中起关作用。动物研究发现经高蔗糖饮食后，与野生型对照小鼠相比肝脏特异性敲除ACC1的小鼠肝脏脂质沉积减少。这提示，肝脏特异性抑制ACC1或通过蛋白翻译后修饰抑制ACC1的酶活性都可能成为NAFLD治疗的靶点。

发明内容

本发明的目的在于克服现有技术的不足，提供一种突变ACC1蛋白及其用途。本发明发现小鼠来源的ACC1蛋白第1523位的赖氨酸(K1523)是ACC1发挥酶活性的关键位点，其丙二酰化水平与非酒精性脂肪性肝病(NAFLD)的发生密切相关。

为实现上述目的，本发明采取的技术方案为：提供一种突变ACC1蛋白，所述突变ACC1蛋白为将野生型ACC1蛋白的第1523位赖氨酸突变为谷氨酸得到，所述野生型ACC1蛋白的氨基酸序列如SEQ ID NO：1所示。

本发明同时提供编码所述突变ACC1蛋白的基因。

作为本发明所述基因的优选实施方式，所述基因的核苷酸序列如SEQ ID NO：4所示。

本发明同时提供一种表达载体，所述表达载体包含所述基因。

本发明同时提供一种宿主细胞，所述宿主细胞包含所述基因或所述表达载体。

本发明同时提供所述突变ACC1蛋白或所述基因在作为诊断非酒精性脂肪性肝病或判断非酒精性脂肪性肝病严重程度的生物标志物中的应用。

本发明还提供所述突变ACC1蛋白、所述基因、所述载体或所述宿主细胞在制备治疗非酒精性脂肪性肝病的抗体和药物研发中的用途。

本发明的有益效果：

1、利用蛋白质组学、生信分析发现并鉴定NAFLD小鼠模型的肝脏中发生显著差异丙二酰化修饰的ACC1蛋白及其丙二酰化位点K1523，说明ACC1蛋白丙二酰化修饰与肝脏脂质沉积有关，将来可作为协助诊断和判断病情严重程度的生物学标记物。

2、利用K1523E突变模拟ACC1蛋白丙二酰化修饰，证明K1523是ACC1发挥酶活性的关键位点，将来可作为抗体制备、NAFLD治疗药物研发的靶点。

附图说明

图1为ACC1 K1523位点丙二酰化修饰在NAFLD发病过程中有关键作用。

图2为K1523是ACC1发挥酶活性的关键位点。

具体实施方式

为更清楚地表述本发明的技术方案，下面结合具体实施例进一步说明，但不能用于限制本发明，此仅是本发明的部分实施例。

除非特别指明，否则基本上按照本领域内熟知的以及在各种参考文献中描述的常规方法进行实施例中描述的实验和方法(例如，分子生物学和核酸化学实验方法)。参见例如，Sambrook等人,Molecular Cloning:A Laboratory Manual,第2版,Cold SpringHarbor Laboratory Press,Cold Spring Harbor,N.Y.(1989)；和Ausubel等人,CurrentProtocols in Molecular Biology,Greene Publishing Associates(1992)，其全部通过引用合并入本文。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。

在本发明中，除非另有说明，否则本文中使用的科学和技术名词具有本领域技术人员所通常理解的含义。并且，本文中所用的分子遗传学、核酸化学和分子生物学相关术语和实验室操作步骤均为相应领域内广泛使用的术语和常规步骤。同时，为了更好地理解本发明，下面提供相关术语的定义和解释。

如本文中所使用的，术语“突变”，当用于描述基因或DNA时，是指基因序列或DNA序列中一个或多个(例如，几个)碱基的添加、缺失和/或置换；当用于描述蛋白质时，是指蛋白质氨基酸序列中一个或多个(例如，几个)氨基酸残基的添加、缺失和/或置换。

实施例1 ACC1 K1523位点丙二酰化修饰对NAFLD发病有关键作用

(1)动物模型发现ACC1 K1523位点丙二酰化修饰在高脂饮食诱导的NAFLD小鼠模型发病过程中有关键作用。

本发明对8周龄雄性C57BL/6小鼠给予高脂饮食(HFD)16周，以普通饮食(CD)作为对照组，通过蛋白质组学及生信分析鉴定肝脏丙二酰化修饰蛋白种类及其修饰位点，GO富集分析结果显示发生丙二酰化修饰的ACC1蛋白与多个糖脂代谢通路(图1A)相关，其K1523位点丙二酰修饰在HFD组显著升高(图1B)，提示ACC1 K1523位点丙二酰化修饰上调对NAFLD的发生有促进作用。

(2)细胞模型发现K1523是ACC1发挥酶活性的关键位点。

本发明将小鼠来源的ACC1蛋白丙二酰化位点上的赖氨酸K突变为谷氨酸E(ACC1K1523E)，从而模拟丙二酰化修饰，对小鼠AML12肝细胞转染携带ACC1 K1523E的质粒，酶活性检测结果提示与野生型(WT)相比，ACC1 K1523E突变蛋白的酶活性显著增强(图2A)，其酶促反应的产物丙二酰辅酶A的水平显著升高(图2B)，肝细胞甘油三酯含量升高(图2C)，肝细胞脂质沉积增加。

表1突变前后酶活性变化

注：数值表示为均值±标准差；增长比率＝(突变后－突变前)/突变前*100％)。

上述结果表明，K1523位点是ACC1发挥酶活性的关键位点，ACC1 K1523丙二酰化修饰对NAFLD发生发展起关键作用，这一发现将为开发ACC1 K1523位点的靶向药物提供研究依据。

最后所应当说明的是，以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制，尽管参照较佳实施例对本发明作了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的实质和范围。

SEQUENCE LISTING

<110> 中山大学附属第三医院

<120> 一种突变ACC1蛋白及其用途

<130> 2021.11.29

<160> 4

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Arg Leu Gly Thr Pro Glu Leu Ser Pro Thr Glu Arg Lys Glu Leu

2150 2155 2160

Glu Ser Lys Leu Lys Glu Arg Glu Glu Phe Leu Ile Pro Ile Tyr

2165 2170 2175

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

2180 2185 2190

Arg Met Gln Glu Lys Gly Val Ile Asn Asp Ile Leu Asp Trp Lys

2195 2200 2205

Thr Ser Arg Thr Phe Phe Tyr Trp Arg Leu Arg Arg Leu Leu Leu

2210 2215 2220

Glu Asp Leu Val Lys Lys Lys Ile His Asn Ala Asn Pro Glu Leu

2225 2230 2235

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

2240 2245 2250

Val Glu Gly Thr Val Lys Ala Tyr Val Trp Asp Asn Asn Lys Asp

2255 2260 2265

Leu Val Glu Trp Leu Glu Lys Gln Leu Thr Glu Glu Asp Gly Val

2270 2275 2280

Arg Ser Val Ile Glu Glu Asn Ile Lys Tyr Ile Ser Arg Asp Tyr

2285 2290 2295

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

2300 2305 2310

Ala Met Asp Ser Ile Val His Met Thr Gln His Ile Ser Pro Thr

2315 2320 2325

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

2330 2335 2340

Ser Thr

2345

<210> 2

<211> 7038

<212> DNA

<213> 人工序列

<400> 2

atggatgaac catctccgtt ggccaaaact ctggagctaa accagcactc ccgattcata 60

attgggtctg tgtctgaaga caactcagaa gatgagatca gtaacctggt gaagctggac 120

ctagaagaga aggagggctc cctgtcacca gcctccgtca gctcagatac actttctgat 180

ttggggatct ctggcttaca ggatggtttg gcctttcaca tgagatccag catgtctggc 240

ttgcacctag taaaacaagg tcgagacaga aagaaaatag actcacaacg agatttcact 300

gtggcttctc cagcagaatt tgttactcgt tttgggggaa ataaagtaat tgagaaggtt 360

cttatcgcca acaatggtat tgcagcagtg aaatgcatgc gatctatccg tcggtggtct 420

tatgaaatgt ttcgaaatga acgtgcaatc cgatttgttg tcatggttac acctgaagac 480

cttaaagcca atgcagaata cattaagatg gcagaccact atgttccagt gcctggagga 540

cccaacaaca acaattacgc aaatgtggag ttgattcttg atattgctaa aaggatacct 600

gtacaagcag tgtgggctgg ctggggtcat gcctctgaga acccgaaact cccagaactg 660

ctcttaaaaa atggcattgc tttcatgggc cctccaagcc aggccatgtg ggctttgggg 720

gataagattg catcttctat tgtggctcaa actgcaggta tcccaactct tccctggagt 780

ggcagtggtc ttcgagtgga ttggcaagaa aatgattttt cgaaacgtat cttaaatgtt 840

ccacaggatc tgtatgagaa aggctatgtg aaggatgtgg atgatggtct gaaggcagct 900

gaggaagttg gctatccagt gatgatcaag gcctcagagg gaggaggagg gaaagggatc 960

agaaaagtta acaatgcaga tgacttccct aacctcttca gacaggttca agctgaagtt 1020

cctggatcac ctatatttgt aatgagacta gcaaaacaat ctcgacatct ggaggtccag 1080

attctggcag atcagtatgg caatgctatt tctttgtttg gtcgtgactg ctctgtgcag 1140

cgcaggcatc agaagatcat tgaagaagct cctgctgcga ttgctacccc agcagtattt 1200

gaacacatgg aacagtgtgc agtgaaactt gccaaaatgg ttggttatgt gagtgctggg 1260

actgtggaat acttgtacag ccaggatgga agcttctact ttttggaact gaaccctcgg 1320

ctacaggttg aacatccttg tacagagatg gtggctgatg tcaatcttcc tgcagcacag 1380

ctccagattg ccatggggat ccctctattt aggatcaagg atattcgtat gatgtatggg 1440

gtatctcctt ggggagatgc tcccattgat tttgaaaatt ctgctcatgt tccttgccca 1500

aggggccacg tgattgctgc tcggatcacc agtgaaaacc cagatgaggg gtttaagccc 1560

agctctggaa cagttcagga acttaatttt cgtagcaata agaacgtttg gggttatttc 1620

agtgttgctg ctgctggagg acttcatgaa tttgctgatt ctcagttcgg gcactgcttt 1680

tcctggggag aaaacaggga ggaagcaatc tcaaatatgg tggtggcact gaaggagctg 1740

tctattcggg gtgactttcg aactacagtg gaatacctca tcaaactgct ggagacagaa 1800

agctttcagc ttaacagaat cgacactggc tggctggaca gactgatcgc agagaaagtg 1860

caggcagagc gacctgacac catgttggga gttgtgtgtg gggctctcca tgtagcagat 1920

gtgagcctga ggaacagcat ctctaacttc cttcactcct tagagagggg tcaagtcctt 1980

cctgctcaca cacttctgaa cacagtagat gttgaactta tctatgaagg aatcaaatat 2040

gtacttaagg tgactcggca gtctcccaac tcctacgtag tgataatgaa tggctcgtgt 2100

gtggaagtgg atgtgcatcg gctgagtgat ggtggcctgc tcttgtctta tgacggcagc 2160

agttacacca catacatgaa ggaagaggtg gacagatatc gaatcacaat tggcaataaa 2220

acctgtgtgt ttgagaagga aaatgaccca tctgtaatgc gctcaccgtc tgctgggaag 2280

ttaatccagt atattgtgga agatggcgga catgtgtttg ctggccagtg ctatgctgag 2340

attgaggtaa tgaagatggt gatgacttta acagctgtag aatctggctg catccattat 2400

gtcaaacgac ctggagcagc acttgaccct ggctgtgtga tagccaaaat gcaactggac 2460

aaccccagta aagttcaaca ggctgagctt cacacgggca gtctaccaca gatccagagc 2520

acagctctca gaggcgagaa gctccatcga gttttccact atgtcctgga taacctggtc 2580

aatgtgatga atggatactg ccttccagac cctttcttca gcagcagggt aaaagactgg 2640

gtagaaagat tgatgaagac tctgagagac ccctccttgc ctctgctaga gctgcaggat 2700

atcatgacca gtgtctctgg ccggatcccc ctcaatgtgg agaagtctat taagaaggaa 2760

atggctcagt atgctagcaa catcacatca gtcctgtgtc agtttcccag ccagcagatt 2820

gccaacatcc tagatagtca tgcagctaca ctgaaccgga aatctgagcg ggaagtcttc 2880

ttcatgaaca cccagagcat tgtccagctg gtgcagaggt accgaagtgg catccgtggc 2940

cacatgaagg ctgtggtgat ggatctgctg cggcagtacc tgcgagtaga gacacagttt 3000

cagaacggcc actacgacaa atgtgtattc gcccttcggg aagagaacaa aagcgacatg 3060

aacaccgtac tgaactacat cttctcccac gctcaggtca ccaaaaagaa tctcctggtg 3120

acaatgctta ttgatcagtt atgtggccgg gaccctacac ttactgatga gctgctaaat 3180

atcctcacag agctaaccca actcagcaag accaccaacg ctaaagtggc gctgcgcgct 3240

cgtcaggttc ttattgcttc ccatttgcca tcatatgagc ttcgccataa ccaagtagag 3300

tctatcttcc tatcagccat tgacatgtat ggacaccagt tttgcattga gaacctgcag 3360

aaactcatcc tctcggaaac atctattttc gatgtcctcc caaacttttt ttaccacagc 3420

aaccaggtgg tgaggatggc agctctggag gtgtatgttc gaagggctta cattgcctat 3480

gaactcaaca gcgtacaaca ccgccagctt aaggacaaca cctgtgtggt ggaatttcag 3540

ttcatgctgc ccacatccca tccaaacaga gggaacatcc ccacgctaaa cagaatgtcc 3600

tttgcctcca acctcaacca ctatggcatg actcatgtag ctagtgtcag cgatgttctg 3660

ttggacaacg ccttcacgcc accttgtcag cggatgggcg gaatggtctc tttccggacc 3720

tttgaagatt ttgtcaggat ctttgatgaa ataatgggct gcttctgtga ctccccaccc 3780

caaagcccca cattcccaga gtctggtcat acttcgctct atgatgaaga caaggtcccc 3840

agggatgaac caatacatat tctgaatgtg gctatcaaga ctgatggcga tattgaggat 3900

gacaggcttg cagctatgtt cagagagttc acccagcaga ataaagctac tttggttgag 3960

catggcatcc ggcgacttac gttcctagtt gcacaaaagg atttcagaaa acaagtcaac 4020

tgtgaggtgg atcagagatt tcatagagaa ttccccaaat ttttcacatt ccgagcaagg 4080

gataagtttg aggaggaccg catttatcga cacctggagc ctgctctggc tttccagtta 4140

gagttgaacc ggatgagaaa ttttgacctt actgccatcc catgtgctaa tcacaagatg 4200

cacctgtacc ttggggctgc taaggtggaa gtaggcacag aagtgactga ctacaggttc 4260

tttgttcgtg cgatcatcag gcactctgat ctggtcacaa aggaagcttc tttcgaatat 4320

ctacaaaatg aaggggaacg actgctcctg gaagctatgg atgaattgga agttgctttt 4380

aataatacaa atgtccgcac tgactgtaac cacatcttcc tcaactttgt gcccacggtc 4440

atcatggacc catcaaagat tgaagaatct gtgcggagca tggtaatgcg ctatggaagt 4500

cggctatgga aattgcgggt cctccaggca gaactgaaaa tcaacattcg cctgacaaca 4560

actggaaaag caattcccat ccgcctcttc ctgacaaacg agtctggcta ctacttggac 4620

atcagcctgt ataaggaagt gactgactcc aggacagcac agatcatgtt tcaggcatat 4680

ggagacaagc agggaccact gcatggaatg ttaattaata ctccatatgt gaccaaagac 4740

cttcttcaat caaagaggtt ccaggcacag tccttaggaa caacatatat atatgatatc 4800

ccagagatgt ttcggcagtc actcatcaaa ctctgggagt ccatgtccac ccaagcattt 4860

cttccttcgc ctcctttgcc ttccgacatc ctgacgtata ctgaactggt gttggatgat 4920

caaggccagc tggtccatat gaacagactt ccaggaggaa atgagattgg catggtagcc 4980

tggaaaatga gccttaaaag ccctgaatat ccagatggcc gagatatcat tgtcatcggc 5040

aatgacatta catatcggat cggttccttt gggcctcagg aggatttgct gtttctcaga 5100

gcttctgaac ttgccagagc agaaggcatc ccacgcatct acgtggcagc gaacagtgga 5160

gctagaattg gacttgcaga agaaatacgc catatgttcc atgtggcctg ggtagatcct 5220

gaagatccct acaagggata caagtattta tatctgacac cccaggatta taagagagtc 5280

agtgccctca attctgtcca ctgtgaacat gtggaggatg aaggggaatc caggtacaag 5340

ataacagata ttatcgggaa agaagaagga cttggagcag agaaccttcg gggttctgga 5400

atgattgctg gggaatcctc attggcttac gatgaggtca tcaccatcag cctggttaca 5460

tgccgggcca ttggtattgg ggcttacctt gtccggctgg gacaaagaac catccaggtt 5520

gagaattctc acttaattct gacaggagca ggtgccctca acaaagtcct tggtcgggaa 5580

gtatacacct ccaacaacca gcttgggggc atccagatta tgcacaacaa tggggttacc 5640

cactccactg tttgtgatga ctttgaggga gtcttcacag tcttacactg gctgtcatac 5700

atgccgaaga gcgtacacag ttcagttcct ctcctgaatt ccaaggatcc tatagataga 5760

atcatcgagt ttgttcccac aaaggcccca tatgatcctc ggtggatgct agcaggccgt 5820

cctcacccaa cccagaaagg ccaatggttg agtgggtttt ttgactatgg atctttctca 5880

gaaatcatgc agccctgggc acagaccgtg gtagttggca gagccaggtt agggggaata 5940

cccgtgggag tagttgctgt agaaacccga acagtggaac tcagtatccc agctgatcct 6000

gcgaacctgg attctgaagc caagataatc cagcaggccg gccaggtttg gttcccagac 6060

tctgcattta agacctatca agctatcaag gactttaacc gtgaagggct acctctaatg 6120

gtctttgcca actggagagg tttctctggt gggatgaaag atatgtatga ccaagtgctc 6180

aagtttggcg cttacattgt ggatggcttg cgggaatgtt cccagcctgt aatggtttac 6240

atcccgcccc aggctgagct tcggggtggt tcttgggttg tgatcgaccc caccatcaac 6300

cctcggcaca tggagatgta cgctgaccga gaaagcaggg gatctgttct ggagccagaa 6360

gggacagtag aaatcaaatt ccgtaaaaag gatctggtga aaaccatgcg tcgggtagat 6420

ccagtttaca tccgcttggc tgagcgattg ggcaccccag agctaagccc cactgagcgg 6480

aaggagctgg agagcaagtt gaaggagcgg gaggagttcc taattcccat ttaccatcag 6540

gtagctgtgc agtttgctga cttgcacgac acaccaggcc ggatgcagga gaagggtgtc 6600

attaacgata tcttggattg gaaaacatcc cgcaccttct tctactggag gctgaggcgc 6660

ctcctgctgg aggacctggt caagaagaaa atccacaatg ccaaccctga gctgactgac 6720

ggccagatcc aggccatgtt gagacgctgg tttgtagaag ttgaaggcac agtgaaggct 6780

tacgtctggg acaataataa ggacctggtg gagtggctgg agaagcaact gacagaggaa 6840

gatggcgtcc gctctgtgat agaggagaac atcaaataca tcagcaggga ctatgtcctg 6900

aagcagatcc gcagcttggt ccaggccaac ccagaagtcg ccatggactc catcgtccac 6960

atgacccagc acatctcacc cacccagcga gcagaagttg taaggatcct ctccactatg 7020

gattcccctt ccacgtag 7038

<210> 3

<211> 2345

<212> PRT

<213> 人工序列

<400> 3

Met Asp Glu Pro Ser Pro Leu Ala Lys Thr Leu Glu Leu Asn Gln His

1 5 10 15

Ser Arg Phe Ile Ile Gly Ser Val Ser Glu Asp Asn Ser Glu Asp Glu

20 25 30

Ile Ser Asn Leu Val Lys Leu Asp Leu Glu Glu Lys Glu Gly Ser Leu

35 40 45

Ser Pro Ala Ser Val Ser Ser Asp Thr Leu Ser Asp Leu Gly Ile Ser

50 55 60

Gly Leu Gln Asp Gly Leu Ala Phe His Met Arg Ser Ser Met Ser Gly

65 70 75 80

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

85 90 95

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

100 105 110

Gly Asn Lys Val Ile Glu Lys Val Leu Ile Ala Asn Asn Gly Ile Ala

115 120 125

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

130 135 140

Arg Asn Glu Arg Ala Ile Arg Phe Val Val Met Val Thr Pro Glu Asp

145 150 155 160

Leu Lys Ala Asn Ala Glu Tyr Ile Lys Met Ala Asp His Tyr Val Pro

165 170 175

Val Pro Gly Gly Pro Asn Asn Asn Asn Tyr Ala Asn Val Glu Leu Ile

180 185 190

Leu Asp Ile Ala Lys Arg Ile Pro Val Gln Ala Val Trp Ala Gly Trp

195 200 205

Gly His Ala Ser Glu Asn Pro Lys Leu Pro Glu Leu Leu Leu Lys Asn

210 215 220

Gly Ile Ala Phe Met Gly Pro Pro Ser Gln Ala Met Trp Ala Leu Gly

225 230 235 240

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

245 250 255

Leu Pro Trp Ser Gly Ser Gly Leu Arg Val Asp Trp Gln Glu Asn Asp

260 265 270

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

275 280 285

Tyr Val Lys Asp Val Asp Asp Gly Leu Lys Ala Ala Glu Glu Val Gly

290 295 300

Tyr Pro Val Met Ile Lys Ala Ser Glu Gly Gly Gly Gly Lys Gly Ile

305 310 315 320

Arg Lys Val Asn Asn Ala Asp Asp Phe Pro Asn Leu Phe Arg Gln Val

325 330 335

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

340 345 350

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

355 360 365

Ala Ile Ser Leu Phe Gly Arg Asp Cys Ser Val Gln Arg Arg His Gln

370 375 380

Lys Ile Ile Glu Glu Ala Pro Ala Ala Ile Ala Thr Pro Ala Val Phe

385 390 395 400

Glu His Met Glu Gln Cys Ala Val Lys Leu Ala Lys Met Val Gly Tyr

405 410 415

Val Ser Ala Gly Thr Val Glu Tyr Leu Tyr Ser Gln Asp Gly Ser Phe

420 425 430

Tyr Phe Leu Glu Leu Asn Pro Arg Leu Gln Val Glu His Pro Cys Thr

435 440 445

Glu Met Val Ala Asp Val Asn Leu Pro Ala Ala Gln Leu Gln Ile Ala

450 455 460

Met Gly Ile Pro Leu Phe Arg Ile Lys Asp Ile Arg Met Met Tyr Gly

465 470 475 480

Val Ser Pro Trp Gly Asp Ala Pro Ile Asp Phe Glu Asn Ser Ala His

485 490 495

Val Pro Cys Pro Arg Gly His Val Ile Ala Ala Arg Ile Thr Ser Glu

500 505 510

Asn Pro Asp Glu Gly Phe Lys Pro Ser Ser Gly Thr Val Gln Glu Leu

515 520 525

Asn Phe Arg Ser Asn Lys Asn Val Trp Gly Tyr Phe Ser Val Ala Ala

530 535 540

Ala Gly Gly Leu His Glu Phe Ala Asp Ser Gln Phe Gly His Cys Phe

545 550 555 560

Ser Trp Gly Glu Asn Arg Glu Glu Ala Ile Ser Asn Met Val Val Ala

565 570 575

Leu Lys Glu Leu Ser Ile Arg Gly Asp Phe Arg Thr Thr Val Glu Tyr

580 585 590

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

595 600 605

Thr Gly Trp Leu Asp Arg Leu Ile Ala Glu Lys Val Gln Ala Glu Arg

610 615 620

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

625 630 635 640

Val Ser Leu Arg Asn Ser Ile Ser Asn Phe Leu His Ser Leu Glu Arg

645 650 655

Gly Gln Val Leu Pro Ala His Thr Leu Leu Asn Thr Val Asp Val Glu

660 665 670

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

675 680 685

Pro Asn Ser Tyr Val Val Ile Met Asn Gly Ser Cys Val Glu Val Asp

690 695 700

Val His Arg Leu Ser Asp Gly Gly Leu Leu Leu Ser Tyr Asp Gly Ser

705 710 715 720

Ser Tyr Thr Thr Tyr Met Lys Glu Glu Val Asp Arg Tyr Arg Ile Thr

725 730 735

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

740 745 750

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

755 760 765

Gly Gly His Val Phe Ala Gly Gln Cys Tyr Ala Glu Ile Glu Val Met

770 775 780

Lys Met Val Met Thr Leu Thr Ala Val Glu Ser Gly Cys Ile His Tyr

785 790 795 800

Val Lys Arg Pro Gly Ala Ala Leu Asp Pro Gly Cys Val Ile Ala Lys

805 810 815

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

820 825 830

Gly Ser Leu Pro Gln Ile Gln Ser Thr Ala Leu Arg Gly Glu Lys Leu

835 840 845

His Arg Val Phe His Tyr Val Leu Asp Asn Leu Val Asn Val Met Asn

850 855 860

Gly Tyr Cys Leu Pro Asp Pro Phe Phe Ser Ser Arg Val Lys Asp Trp

865 870 875 880

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

885 890 895

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

900 905 910

Val Glu Lys Ser Ile Lys Lys Glu Met Ala Gln Tyr Ala Ser Asn Ile

915 920 925

Thr Ser Val Leu Cys Gln Phe Pro Ser Gln Gln Ile Ala Asn Ile Leu

930 935 940

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

945 950 955 960

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

965 970 975

Gly Ile Arg Gly His Met Lys Ala Val Val Met Asp Leu Leu Arg Gln

980 985 990

Tyr Leu Arg Val Glu Thr Gln Phe Gln Asn Gly His Tyr Asp Lys Cys

995 1000 1005

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

1010 1015 1020

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

1025 1030 1035

Leu Val Thr Met Leu Ile Asp Gln Leu Cys Gly Arg Asp Pro Thr

1040 1045 1050

Leu Thr Asp Glu Leu Leu Asn Ile Leu Thr Glu Leu Thr Gln Leu

1055 1060 1065

Ser Lys Thr Thr Asn Ala Lys Val Ala Leu Arg Ala Arg Gln Val

1070 1075 1080

Leu Ile Ala Ser His Leu Pro Ser Tyr Glu Leu Arg His Asn Gln

1085 1090 1095

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

1100 1105 1110

Phe Cys Ile Glu Asn Leu Gln Lys Leu Ile Leu Ser Glu Thr Ser

1115 1120 1125

Ile Phe Asp Val Leu Pro Asn Phe Phe Tyr His Ser Asn Gln Val

1130 1135 1140

Val Arg Met Ala Ala Leu Glu Val Tyr Val Arg Arg Ala Tyr Ile

1145 1150 1155

Ala Tyr Glu Leu Asn Ser Val Gln His Arg Gln Leu Lys Asp Asn

1160 1165 1170

Thr Cys Val Val Glu Phe Gln Phe Met Leu Pro Thr Ser His Pro

1175 1180 1185

Asn Arg Gly Asn Ile Pro Thr Leu Asn Arg Met Ser Phe Ala Ser

1190 1195 1200

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

1205 1210 1215

Val Leu Leu Asp Asn Ala Phe Thr Pro Pro Cys Gln Arg Met Gly

1220 1225 1230

Gly Met Val Ser Phe Arg Thr Phe Glu Asp Phe Val Arg Ile Phe

1235 1240 1245

Asp Glu Ile Met Gly Cys Phe Cys Asp Ser Pro Pro Gln Ser Pro

1250 1255 1260

Thr Phe Pro Glu Ser Gly His Thr Ser Leu Tyr Asp Glu Asp Lys

1265 1270 1275

Val Pro Arg Asp Glu Pro Ile His Ile Leu Asn Val Ala Ile Lys

1280 1285 1290

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

1295 1300 1305

Glu Phe Thr Gln Gln Asn Lys Ala Thr Leu Val Glu His Gly Ile

1310 1315 1320

Arg Arg Leu Thr Phe Leu Val Ala Gln Lys Asp Phe Arg Lys Gln

1325 1330 1335

Val Asn Cys Glu Val Asp Gln Arg Phe His Arg Glu Phe Pro Lys

1340 1345 1350

Phe Phe Thr Phe Arg Ala Arg Asp Lys Phe Glu Glu Asp Arg Ile

1355 1360 1365

Tyr Arg His Leu Glu Pro Ala Leu Ala Phe Gln Leu Glu Leu Asn

1370 1375 1380

Arg Met Arg Asn Phe Asp Leu Thr Ala Ile Pro Cys Ala Asn His

1385 1390 1395

Lys Met His Leu Tyr Leu Gly Ala Ala Lys Val Glu Val Gly Thr

1400 1405 1410

Glu Val Thr Asp Tyr Arg Phe Phe Val Arg Ala Ile Ile Arg His

1415 1420 1425

Ser Asp Leu Val Thr Lys Glu Ala Ser Phe Glu Tyr Leu Gln Asn

1430 1435 1440

Glu Gly Glu Arg Leu Leu Leu Glu Ala Met Asp Glu Leu Glu Val

1445 1450 1455

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

1460 1465 1470

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

1475 1480 1485

Glu Ser Val Arg Ser Met Val Met Arg Tyr Gly Ser Arg Leu Trp

1490 1495 1500

Lys Leu Arg Val Leu Gln Ala Glu Leu Lys Ile Asn Ile Arg Leu

1505 1510 1515

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

1520 1525 1530

Glu Ser Gly Tyr Tyr Leu Asp Ile Ser Leu Tyr Lys Glu Val Thr

1535 1540 1545

Asp Ser Arg Thr Ala Gln Ile Met Phe Gln Ala Tyr Gly Asp Lys

1550 1555 1560

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

1565 1570 1575

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

1580 1585 1590

Thr Thr Tyr Ile Tyr Asp Ile Pro Glu Met Phe Arg Gln Ser Leu

1595 1600 1605

Ile Lys Leu Trp Glu Ser Met Ser Thr Gln Ala Phe Leu Pro Ser

1610 1615 1620

Pro Pro Leu Pro Ser Asp Ile Leu Thr Tyr Thr Glu Leu Val Leu

1625 1630 1635

Asp Asp Gln Gly Gln Leu Val His Met Asn Arg Leu Pro Gly Gly

1640 1645 1650

Asn Glu Ile Gly Met Val Ala Trp Lys Met Ser Leu Lys Ser Pro

1655 1660 1665

Glu Tyr Pro Asp Gly Arg Asp Ile Ile Val Ile Gly Asn Asp Ile

1670 1675 1680

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

1685 1690 1695

Leu Arg Ala Ser Glu Leu Ala Arg Ala Glu Gly Ile Pro Arg Ile

1700 1705 1710

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

1715 1720 1725

Ile Arg His Met Phe His Val Ala Trp Val Asp Pro Glu Asp Pro

1730 1735 1740

Tyr Lys Gly Tyr Lys Tyr Leu Tyr Leu Thr Pro Gln Asp Tyr Lys

1745 1750 1755

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

1760 1765 1770

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

1775 1780 1785

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

1790 1795 1800

Gly Glu Ser Ser Leu Ala Tyr Asp Glu Val Ile Thr Ile Ser Leu

1805 1810 1815

Val Thr Cys Arg Ala Ile Gly Ile Gly Ala Tyr Leu Val Arg Leu

1820 1825 1830

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

1835 1840 1845

Gly Ala Gly Ala Leu Asn Lys Val Leu Gly Arg Glu Val Tyr Thr

1850 1855 1860

Ser Asn Asn Gln Leu Gly Gly Ile Gln Ile Met His Asn Asn Gly

1865 1870 1875

Val Thr His Ser Thr Val Cys Asp Asp Phe Glu Gly Val Phe Thr

1880 1885 1890

Val Leu His Trp Leu Ser Tyr Met Pro Lys Ser Val His Ser Ser

1895 1900 1905

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

1910 1915 1920

Phe Val Pro Thr Lys Ala Pro Tyr Asp Pro Arg Trp Met Leu Ala

1925 1930 1935

Gly Arg Pro His Pro Thr Gln Lys Gly Gln Trp Leu Ser Gly Phe

1940 1945 1950

Phe Asp Tyr Gly Ser Phe Ser Glu Ile Met Gln Pro Trp Ala Gln

1955 1960 1965

Thr Val Val Val Gly Arg Ala Arg Leu Gly Gly Ile Pro Val Gly

1970 1975 1980

Val Val Ala Val Glu Thr Arg Thr Val Glu Leu Ser Ile Pro Ala

1985 1990 1995

Asp Pro Ala Asn Leu Asp Ser Glu Ala Lys Ile Ile Gln Gln Ala

2000 2005 2010

Gly Gln Val Trp Phe Pro Asp Ser Ala Phe Lys Thr Tyr Gln Ala

2015 2020 2025

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

2030 2035 2040

Asn Trp Arg Gly Phe Ser Gly Gly Met Lys Asp Met Tyr Asp Gln

2045 2050 2055

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

2060 2065 2070

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

2075 2080 2085

Gly Gly Ser Trp Val Val Ile Asp Pro Thr Ile Asn Pro Arg His

2090 2095 2100

Met Glu Met Tyr Ala Asp Arg Glu Ser Arg Gly Ser Val Leu Glu

2105 2110 2115

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

2120 2125 2130

Lys Thr Met Arg Arg Val Asp Pro Val Tyr Ile Arg Leu Ala Glu

2135 2140 2145

Arg Leu Gly Thr Pro Glu Leu Ser Pro Thr Glu Arg Lys Glu Leu

2150 2155 2160

Glu Ser Lys Leu Lys Glu Arg Glu Glu Phe Leu Ile Pro Ile Tyr

2165 2170 2175

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

2180 2185 2190

Arg Met Gln Glu Lys Gly Val Ile Asn Asp Ile Leu Asp Trp Lys

2195 2200 2205

Thr Ser Arg Thr Phe Phe Tyr Trp Arg Leu Arg Arg Leu Leu Leu

2210 2215 2220

Glu Asp Leu Val Lys Lys Lys Ile His Asn Ala Asn Pro Glu Leu

2225 2230 2235

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

2240 2245 2250

Val Glu Gly Thr Val Lys Ala Tyr Val Trp Asp Asn Asn Lys Asp

2255 2260 2265

Leu Val Glu Trp Leu Glu Lys Gln Leu Thr Glu Glu Asp Gly Val

2270 2275 2280

Arg Ser Val Ile Glu Glu Asn Ile Lys Tyr Ile Ser Arg Asp Tyr

2285 2290 2295

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

2300 2305 2310

Ala Met Asp Ser Ile Val His Met Thr Gln His Ile Ser Pro Thr

2315 2320 2325

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

2330 2335 2340

Ser Thr

2345

<210> 4

<211> 7038

<212> DNA

<213> 人工序列

<400> 4

atggatgaac catctccgtt ggccaaaact ctggagctaa accagcactc ccgattcata 60

attgggtctg tgtctgaaga caactcagaa gatgagatca gtaacctggt gaagctggac 120

ctagaagaga aggagggctc cctgtcacca gcctccgtca gctcagatac actttctgat 180

ttggggatct ctggcttaca ggatggtttg gcctttcaca tgagatccag catgtctggc 240

ttgcacctag taaaacaagg tcgagacaga aagaaaatag actcacaacg agatttcact 300

gtggcttctc cagcagaatt tgttactcgt tttgggggaa ataaagtaat tgagaaggtt 360

cttatcgcca acaatggtat tgcagcagtg aaatgcatgc gatctatccg tcggtggtct 420

tatgaaatgt ttcgaaatga acgtgcaatc cgatttgttg tcatggttac acctgaagac 480

cttaaagcca atgcagaata cattaagatg gcagaccact atgttccagt gcctggagga 540

cccaacaaca acaattacgc aaatgtggag ttgattcttg atattgctaa aaggatacct 600

gtacaagcag tgtgggctgg ctggggtcat gcctctgaga acccgaaact cccagaactg 660

ctcttaaaaa atggcattgc tttcatgggc cctccaagcc aggccatgtg ggctttgggg 720

gataagattg catcttctat tgtggctcaa actgcaggta tcccaactct tccctggagt 780

ggcagtggtc ttcgagtgga ttggcaagaa aatgattttt cgaaacgtat cttaaatgtt 840

ccacaggatc tgtatgagaa aggctatgtg aaggatgtgg atgatggtct gaaggcagct 900

gaggaagttg gctatccagt gatgatcaag gcctcagagg gaggaggagg gaaagggatc 960

agaaaagtta acaatgcaga tgacttccct aacctcttca gacaggttca agctgaagtt 1020

cctggatcac ctatatttgt aatgagacta gcaaaacaat ctcgacatct ggaggtccag 1080

attctggcag atcagtatgg caatgctatt tctttgtttg gtcgtgactg ctctgtgcag 1140

cgcaggcatc agaagatcat tgaagaagct cctgctgcga ttgctacccc agcagtattt 1200

gaacacatgg aacagtgtgc agtgaaactt gccaaaatgg ttggttatgt gagtgctggg 1260

actgtggaat acttgtacag ccaggatgga agcttctact ttttggaact gaaccctcgg 1320

ctacaggttg aacatccttg tacagagatg gtggctgatg tcaatcttcc tgcagcacag 1380

ctccagattg ccatggggat ccctctattt aggatcaagg atattcgtat gatgtatggg 1440

gtatctcctt ggggagatgc tcccattgat tttgaaaatt ctgctcatgt tccttgccca 1500

aggggccacg tgattgctgc tcggatcacc agtgaaaacc cagatgaggg gtttaagccc 1560

agctctggaa cagttcagga acttaatttt cgtagcaata agaacgtttg gggttatttc 1620

agtgttgctg ctgctggagg acttcatgaa tttgctgatt ctcagttcgg gcactgcttt 1680

tcctggggag aaaacaggga ggaagcaatc tcaaatatgg tggtggcact gaaggagctg 1740

tctattcggg gtgactttcg aactacagtg gaatacctca tcaaactgct ggagacagaa 1800

agctttcagc ttaacagaat cgacactggc tggctggaca gactgatcgc agagaaagtg 1860

caggcagagc gacctgacac catgttggga gttgtgtgtg gggctctcca tgtagcagat 1920

gtgagcctga ggaacagcat ctctaacttc cttcactcct tagagagggg tcaagtcctt 1980

cctgctcaca cacttctgaa cacagtagat gttgaactta tctatgaagg aatcaaatat 2040

gtacttaagg tgactcggca gtctcccaac tcctacgtag tgataatgaa tggctcgtgt 2100

gtggaagtgg atgtgcatcg gctgagtgat ggtggcctgc tcttgtctta tgacggcagc 2160

agttacacca catacatgaa ggaagaggtg gacagatatc gaatcacaat tggcaataaa 2220

acctgtgtgt ttgagaagga aaatgaccca tctgtaatgc gctcaccgtc tgctgggaag 2280

ttaatccagt atattgtgga agatggcgga catgtgtttg ctggccagtg ctatgctgag 2340

attgaggtaa tgaagatggt gatgacttta acagctgtag aatctggctg catccattat 2400

gtcaaacgac ctggagcagc acttgaccct ggctgtgtga tagccaaaat gcaactggac 2460

aaccccagta aagttcaaca ggctgagctt cacacgggca gtctaccaca gatccagagc 2520

acagctctca gaggcgagaa gctccatcga gttttccact atgtcctgga taacctggtc 2580

aatgtgatga atggatactg ccttccagac cctttcttca gcagcagggt aaaagactgg 2640

gtagaaagat tgatgaagac tctgagagac ccctccttgc ctctgctaga gctgcaggat 2700

atcatgacca gtgtctctgg ccggatcccc ctcaatgtgg agaagtctat taagaaggaa 2760

atggctcagt atgctagcaa catcacatca gtcctgtgtc agtttcccag ccagcagatt 2820

gccaacatcc tagatagtca tgcagctaca ctgaaccgga aatctgagcg ggaagtcttc 2880

ttcatgaaca cccagagcat tgtccagctg gtgcagaggt accgaagtgg catccgtggc 2940

cacatgaagg ctgtggtgat ggatctgctg cggcagtacc tgcgagtaga gacacagttt 3000

cagaacggcc actacgacaa atgtgtattc gcccttcggg aagagaacaa aagcgacatg 3060

aacaccgtac tgaactacat cttctcccac gctcaggtca ccaaaaagaa tctcctggtg 3120

acaatgctta ttgatcagtt atgtggccgg gaccctacac ttactgatga gctgctaaat 3180

atcctcacag agctaaccca actcagcaag accaccaacg ctaaagtggc gctgcgcgct 3240

cgtcaggttc ttattgcttc ccatttgcca tcatatgagc ttcgccataa ccaagtagag 3300

tctatcttcc tatcagccat tgacatgtat ggacaccagt tttgcattga gaacctgcag 3360

aaactcatcc tctcggaaac atctattttc gatgtcctcc caaacttttt ttaccacagc 3420

aaccaggtgg tgaggatggc agctctggag gtgtatgttc gaagggctta cattgcctat 3480

gaactcaaca gcgtacaaca ccgccagctt aaggacaaca cctgtgtggt ggaatttcag 3540

ttcatgctgc ccacatccca tccaaacaga gggaacatcc ccacgctaaa cagaatgtcc 3600

tttgcctcca acctcaacca ctatggcatg actcatgtag ctagtgtcag cgatgttctg 3660

ttggacaacg ccttcacgcc accttgtcag cggatgggcg gaatggtctc tttccggacc 3720

tttgaagatt ttgtcaggat ctttgatgaa ataatgggct gcttctgtga ctccccaccc 3780

caaagcccca cattcccaga gtctggtcat acttcgctct atgatgaaga caaggtcccc 3840

agggatgaac caatacatat tctgaatgtg gctatcaaga ctgatggcga tattgaggat 3900

gacaggcttg cagctatgtt cagagagttc acccagcaga ataaagctac tttggttgag 3960

catggcatcc ggcgacttac gttcctagtt gcacaaaagg atttcagaaa acaagtcaac 4020

tgtgaggtgg atcagagatt tcatagagaa ttccccaaat ttttcacatt ccgagcaagg 4080

gataagtttg aggaggaccg catttatcga cacctggagc ctgctctggc tttccagtta 4140

gagttgaacc ggatgagaaa ttttgacctt actgccatcc catgtgctaa tcacaagatg 4200

cacctgtacc ttggggctgc taaggtggaa gtaggcacag aagtgactga ctacaggttc 4260

tttgttcgtg cgatcatcag gcactctgat ctggtcacaa aggaagcttc tttcgaatat 4320

ctacaaaatg aaggggaacg actgctcctg gaagctatgg atgaattgga agttgctttt 4380

aataatacaa atgtccgcac tgactgtaac cacatcttcc tcaactttgt gcccacggtc 4440

atcatggacc catcaaagat tgaagaatct gtgcggagca tggtaatgcg ctatggaagt 4500

cggctatgga aattgcgggt cctccaggca gaactgaaaa tcaacattcg cctgacaaca 4560

actggagaag caattcccat ccgcctcttc ctgacaaacg agtctggcta ctacttggac 4620

atcagcctgt ataaggaagt gactgactcc aggacagcac agatcatgtt tcaggcatat 4680

ggagacaagc agggaccact gcatggaatg ttaattaata ctccatatgt gaccaaagac 4740

cttcttcaat caaagaggtt ccaggcacag tccttaggaa caacatatat atatgatatc 4800

ccagagatgt ttcggcagtc actcatcaaa ctctgggagt ccatgtccac ccaagcattt 4860

cttccttcgc ctcctttgcc ttccgacatc ctgacgtata ctgaactggt gttggatgat 4920

caaggccagc tggtccatat gaacagactt ccaggaggaa atgagattgg catggtagcc 4980

tggaaaatga gccttaaaag ccctgaatat ccagatggcc gagatatcat tgtcatcggc 5040

aatgacatta catatcggat cggttccttt gggcctcagg aggatttgct gtttctcaga 5100

gcttctgaac ttgccagagc agaaggcatc ccacgcatct acgtggcagc gaacagtgga 5160

gctagaattg gacttgcaga agaaatacgc catatgttcc atgtggcctg ggtagatcct 5220

gaagatccct acaagggata caagtattta tatctgacac cccaggatta taagagagtc 5280

agtgccctca attctgtcca ctgtgaacat gtggaggatg aaggggaatc caggtacaag 5340

ataacagata ttatcgggaa agaagaagga cttggagcag agaaccttcg gggttctgga 5400

atgattgctg gggaatcctc attggcttac gatgaggtca tcaccatcag cctggttaca 5460

tgccgggcca ttggtattgg ggcttacctt gtccggctgg gacaaagaac catccaggtt 5520

gagaattctc acttaattct gacaggagca ggtgccctca acaaagtcct tggtcgggaa 5580

gtatacacct ccaacaacca gcttgggggc atccagatta tgcacaacaa tggggttacc 5640

cactccactg tttgtgatga ctttgaggga gtcttcacag tcttacactg gctgtcatac 5700

atgccgaaga gcgtacacag ttcagttcct ctcctgaatt ccaaggatcc tatagataga 5760

atcatcgagt ttgttcccac aaaggcccca tatgatcctc ggtggatgct agcaggccgt 5820

cctcacccaa cccagaaagg ccaatggttg agtgggtttt ttgactatgg atctttctca 5880

gaaatcatgc agccctgggc acagaccgtg gtagttggca gagccaggtt agggggaata 5940

cccgtgggag tagttgctgt agaaacccga acagtggaac tcagtatccc agctgatcct 6000

gcgaacctgg attctgaagc caagataatc cagcaggccg gccaggtttg gttcccagac 6060

tctgcattta agacctatca agctatcaag gactttaacc gtgaagggct acctctaatg 6120

gtctttgcca actggagagg tttctctggt gggatgaaag atatgtatga ccaagtgctc 6180

aagtttggcg cttacattgt ggatggcttg cgggaatgtt cccagcctgt aatggtttac 6240

atcccgcccc aggctgagct tcggggtggt tcttgggttg tgatcgaccc caccatcaac 6300

cctcggcaca tggagatgta cgctgaccga gaaagcaggg gatctgttct ggagccagaa 6360

gggacagtag aaatcaaatt ccgtaaaaag gatctggtga aaaccatgcg tcgggtagat 6420

ccagtttaca tccgcttggc tgagcgattg ggcaccccag agctaagccc cactgagcgg 6480

aaggagctgg agagcaagtt gaaggagcgg gaggagttcc taattcccat ttaccatcag 6540

gtagctgtgc agtttgctga cttgcacgac acaccaggcc ggatgcagga gaagggtgtc 6600

attaacgata tcttggattg gaaaacatcc cgcaccttct tctactggag gctgaggcgc 6660

ctcctgctgg aggacctggt caagaagaaa atccacaatg ccaaccctga gctgactgac 6720

ggccagatcc aggccatgtt gagacgctgg tttgtagaag ttgaaggcac agtgaaggct 6780

tacgtctggg acaataataa ggacctggtg gagtggctgg agaagcaact gacagaggaa 6840

gatggcgtcc gctctgtgat agaggagaac atcaaataca tcagcaggga ctatgtcctg 6900

aagcagatcc gcagcttggt ccaggccaac ccagaagtcg ccatggactc catcgtccac 6960

atgacccagc acatctcacc cacccagcga gcagaagttg taaggatcct ctccactatg 7020

gattcccctt ccacgtag 7038

Claims

1.一种突变ACC1蛋白，其特征在于，所述突变ACC1蛋白为将野生型ACC1蛋白的第1523位赖氨酸突变为谷氨酸得到，所述野生型ACC1蛋白的氨基酸序列如SEQ ID NO：1所示。

2.编码权利要求1所述突变ACC1蛋白的基因。

3.根据权利要求2所述的基因，其特征在于，所述基因的核苷酸序列如SEQ ID NO：4所示。

4.一种表达载体，其特征在于，所述表达载体包含权利要求2所述基因。