XJ-160病毒感染性全基因组cDNA克隆

摘要：

本发明提供了一种XJ-160病毒感染性全基因组cDNA克隆pBR-XJ160，该克隆由骨架质粒pBR、含11620个碱基对的病毒基因组全长cDNA、基因组5’端的SP6启动子序列和基因组3’端的35个连续的A组成。其中所述的骨架质粒pBR由线性化的质粒pBR322与含单酶切位点的连接子连接而成；所述的病毒基因组全长cDNA由三个cDNA组装片段通过所述单酶切位点插入骨架质粒pBR组装而成，三个组装片段顺次连接，覆盖了XJ-160病毒的全长基因组；所述的SP6启动子序列位于基因组5’端上游紧邻第一个核苷酸A，所述的35个连续的A紧邻基因组3’末端核甘酸C，该克隆可在宿主菌中稳定传代，其RNA转录体转染宿主细胞后可以产生恢复病毒(BR-XJ160)，并可以通过遗传标记的鉴定区分恢复病毒与亲本病毒。恢复病毒(BR-XJ160)较好的重现了XJ-160病毒的各种生物学特征，该感染性全基因组cDNA克隆是理想的XJ-160病毒的反向遗传学系统。

专利类型：发明专利

申请/专利号：CN200310115457.9

申请日期：2003-11-25

公开/公告号：CN1621527

公开/公告日：2005-06-01

主分类号：C12P19/34

申请/专利权人:中国疾病预防控制中心病毒病预防控制所

发明/设计人:梁国栋;杨益良;付士红;何海怀;侯云德

主申请人地址:100052北京市宣武区迎新路100号

专利代理机构:北京北新智诚知识产权代理有限公司

代理人:郭佩兰

国别省市代码:北京;11

权利要求：

1.一种XJ-160病毒的感染性全基因组cDNA克隆，其特征在于：该克隆由低拷贝骨架质粒pBR、病毒基因组全长cDNA片段、基因组5’端的SP6启动子序列和基因组3’端的35个连续的A组成。其中所述的骨架质粒pBR是线性化的质粒pBR322与含单酶切位点的连接子连接而成；所述的病毒基因组全长cDNA片段含有11620个核甘酸，通过所述单酶切位点将覆盖病毒全长基因组的cDNA片段插入到骨架质粒pBR中；所述的SP6启动子序列位于基因组5’端上游紧邻第一个核苷酸A，所述的35个连续的A紧邻基因组3’末端核甘酸C。该感染性全基因组cDNA克隆序列为：1ATTGACGGCGTAGTACACACTATTGAATCAAACAGCCGACCAATAGCACT51ACCATCATAATGGAGAGGCCTGTAGTTAACGTAGACGTAGACCCCCAGAG101TCCGTTTGTCGCTCAACTGCAAAAGAGCTTCCCGCAATTCGAGGTAGTAG151CACAGCAGGCCACACCAAATGACCATGCTAATGCCAGAGCATTTTCGCAT201CTGGCTAGTAAATTAATCGAGCTGGAGGTTCCTACCACAGCGACGATTTT251GGACATAGGCAGCGCACCGGCTCGTAGAATGTTTTCCGAGCACCACTATC301ACTGCGTCTGCCCTATGCGGAGCCCCGAAGATCCCGACCGTATGATGAAA351TACGCCAATAAGTTGGCGGAGAAGGCAAATAAGATTACTAATAAAAATTT401GCATGAGAAGATTAAAGACCTCCGGATCGTACTCGATACTCCGGATGCTG451AGACACCGTCGCTCTGCTTCCATAATGACGTTACCTGCAGTACGCGTGCA501GAGTACTCCGTTATGCAAGATGTGTATATTAATGCACCCGGAACTATTTA551CCATCAAGCTATGAAAGGCGTGCGGACACTTTACTGGATTGGGTTTGACA601CCACTCAGTTCATGTTCTCGGCTATGGCAGGATCATATCCTGCCTATAAT651ACTAACTGGGCCGATGAAAAAGTCCTTGAAGCACGCAATATTGGACTCTG701TAGTACCAAGTTGAGCGAAGGCCGTATAGGAAAGTTGTCAATAATGAGGA751AAAAGGAGTTGAAGCCCGGGTCACGGGTCTACTTCTCAGTGGGATCAACA801CTCTACCCAGAATATAGGGACAGCTTACAGAGCTGGCACCTCCCATCAGT851GTTCCATCTGAAAGGAAAGCAATCGTATACATGCCGCTGTGATACAGTGG901TAAGTTGCGAAGGCTACGTAGTGAAGAAGATCACTATTAGTCCCGGGATT951ACGGGAGAAACCGTGGGATACGCGGTTACAAACAATAGCGAGGGTTTCTT1001GCTATGTAAAGTTACTGACACAGTAAAAGGGGAACGGGTCTCGTTCCCCG1051TGTGCACGTATATCCCGGCCACCATATGCGACCAGATGACAGGTATAATG1101GCCACGGATATTTCACCTGACGATGCACAGAAGCTTCTGGTTGGGCTCAA1151CCAGAGGATTGTCATTAACGGTAAAACCAACAGGAACACTAACACCATGC1201AAAACTACCTTCTACCGATTATAGCACAAGGCTTCAGCAAGTGGGCTAAA1251GAGCGCAAAGAAGATCTTGATAATGAAAAGAAGCTGGGCACTAGGGAGCG1301TAAGCTTATTTACGGGTGTCTATGGGCGGTTCGTACTAAGAAAGTGCACT1351CGTTTTACCGCCCGCCCGGAACGCAGACCAGCGTGAAAGTCCCGGCATCT1401TTTAGCGCTTTTCCAATGTCATCTGTATGGACAACCTCACTACCCATGTC1451GCTGAGGCAGAAGATAAAATTGGTACTACAACCGAAAAAGGAGGAGAAAT1501TACTGCAGGTCTCAGAAGAGTTAGTTGCGGAGGCTAAAGCGGCCTTTGAA1551GATGCACAGGAGGAGATCAGAGCGGAGCAACTCCGTGAAGCACTTCCACC1601ACTGGTAGCAGACAAAGGTATTGAAGCCGCTGCAGAAGTCGTCTGTGAAG1651TGGAAGGGCTTCAAGCTGATATAGGAGCAGCTCTTGTTGAGACGCCACGA1701GGGCATGTAAGGATTATACCTCAAGTAACAGACCGCATGATTGGGCAGTA1751CATCGTGGTCTCACCAACCTCCGTGCTTAAGAACGCCAAATTAACACCTG1801TTCACCCTTTGGCTGACCAAGTCAAAATTATAACACATTCAGGGAGGACA1851GGAAGATTCGCGGTGGAGCCGTATGATGCTAAAGTGTTGATGCCAGCAGG1901TAGCGCCGTTCCATGGCCTGAGTTCCTGGCGTTAAGTGAAAGCGCCACGC1951TAGTGTATAACGAAAGAGAATTTGTCAACCGCAAACTTTACCATATCGCC2001ATACATGGTCCCGCGAAGAATACTGAAGAGGAGCAGTATAAAGTCACTAA2051AGCCGAACTCGCAGAAACAGAGTATGTATTTGATGTCGACAAGAAGCGTT2101GCGTTAAGAAGGAAGAGGCCTCGGGGCTGGTTCTCTCGGGAGAACTAACT2151AACCCACCGTATCATGAAATGGCGCTTGAGGGGCTGAAGACTCGACCCGC2201TGTTCCGTATAAGGTCGAAACAATAGGAGTAATAGGCACACCAGGATCAG2251GCAAGTCTGCGATTATTAAATCGACCGTTACCGTACGAGATCTTGTTACC2301AGCGGAAAGAAGGAAAACTGCCGTGAAATTGAAACTGACGTGTTGAGGTT2351GAGAGGTATGCAGATCACGTCAAGGACGGTAGACTCAGTCATGCTTAACG2401GATGCCATAAAGCCGTTGAGGTGCTATACGTTGATGAAGCATTTGCGTGC2451CATGCTGGTACGTTGCTTGCCTTGATCGCTATTGTTAGACCCCGAAAGAA2501GGTAGTACTTCGCGGGGATCCTAAGCAGTGTGGTTTTTTCAATATGATGC2551AGCTCAAAGTACATTTTAATCACCCTGAAAAAGATATATGTACTAAGACG2601TTTTACAAATTCATCTCTCGACGCTGCACGCAACCAGTAACGGCGATTGT2651TTCAACACTTCATTACGACGGAAAGATGAAGACTACAAACCCCTGCAAGA2701AGAGCATTGAGATAGATATTACAGGTACTACGAAGCCGAAGCCCGGGGAC2751CTCGTTTTGACGTGCTTCCGCGGATGGGTCAAGCAGCTACAGATCGATTA2801CGCGGGAAATGAAGTGATGACGGCTGCTGCCTCGCAGGGACTGACTAGAA2851AGGGTGTTTACGCCGTTCGGCAAAAAGTTAATGAGAATCCACTGTACGCG2901ATTACGTCGGAGCACGTGAACGTGTTACTCACCCGTACTGAGGATAGATT2951AGTGTGGAAGACCTTACAGGGCGATCCATGGATCAAACAACTTACTAACA3001TTCCGAAAGGAAACTTCCAAGCTACTATTGAGGACTGGGAAGCTGAACAT3051AAGGGGATCATCGCTGCAATAAACAGCCCAACCCCTCGTATAAACCCGTT3101CAGCTGTAAGACTAATGTGTGCTGGGCGAAGGCACTGGAACCGATACTGG3151CCACAGCCGGTATTGTCCTCACCGGTTGCCAGTGGAGTGAGCTGTTTCCA3201CAGTTCGTAGATGATAAACCCCACTCAGCTATCTACGCCCTAGATGTGAT3251TTGTATTAAGTTCTTTGGTATGGATCTGACAAGCGGTCTATTTTCAAAGC3301AGAGCATCCCTCTGACGTACCACCCTGCGGACTCTGCAAGGCCAGTGGCG3351CATTGGGACAACAGTCCAGGAACCCGAAAGTATGGATACGATCATGCGGT3401TGCTGCCGAATTGTCTCGTAGATTCCCGGTGTTCCAACTTGCTGGAAAAG3451GCACACAGCTTGACTTGCAGACTGGTAGAACCAGAGTCGTTTCCGCGCAG3501TGTAACTTGGTCCCAGTGAACCGTAACCTCCCGCACGCTCTTGTCCCCGA3551GTATAAAGAGAAACAACCCGGCCCGATTAAAAATTTTTTAAATCAGTTTA3601AGCATCACTCCATACTTGTAGTATCAGAAACGAAAATCGAAGTTCCCAAT3651AAGCGCATCGAATGGATTGCACCGCTTGGCATAGCTGGTGCAGATAAGAG3701CTACAACCTGGCCTTCGGATTTCCACCGCAGGCACGGTATGATATGGTGT3751TTATCAATATAGGAACAAAATATAGAAACCACCATTTTCAACAGTGTGAA3801GACCATGCGGCGACTTTGAAGACTCTTTCCCGCTCGGCTCTGAATTGCCT3851CAACCCTGGAGGCACCTTAGTGGTGAAATCCTATGGATATGCTGATCGCA3901ATAGCGAGGACGTAGTCACCGCACTTGCCAGGAAGTTTGTTAGAGTGTCT3951GCGGCCAGGCCAGAGTGCGTCTCAAGTAACACAGAAATGTACCTAATCTT4001TCGGCAATTAGATAATAGCCGTACACGGCAGTTCACTCCACATCATTTGA4051ACTGTGTAATCTCGTCGGTGTATGAAGGCACGAGAGAAGGAGTCGGAGCT4101GCGCCATCCTACCGTGTGAAACGGGAAAATATTGCAGACTGCCATGAGGA4151AGCAATCGTCAACGCCGCTAACTCGCTGGGTAAACCAGGTGAAGGAGTTT4201GCCGCGCCGTCTACAAGCGTTGGCCGAGCAGCTTTATGGATTCCGCCACA4251GAAACGGGTACGGCTAAATTAACTGTAAGCCAAGGAATGAAAGTGATACA4301CGCGGTCGGCCCTGACTTCCGTAAGTATCCTGAGGCGGAAGCTTTGAAGC4351TGCTGCAAAACGCTTACCATGCAGTGGCGGACTTGGTTAACAAACACAAC4401ATTAAGTCCATTGCTATCCCGCTACTATCAACAGGTATATATGCAGCTGG4451TAAGGATCGCTTGGAAGTTTCGCTCAATTGCCTGACCACCGCACTAGACA4501GGACCGATGCAGATGTAACTATTTATTGTTTGGATAAGAAATGGAAAGAG4551AGAATTGACGCGGTGTTGCAACTTAAGGAGTCAGTGACAGAACTGAAGGA4601CGAGGACATGGAAATCGATGACGAATTGGTATGGATTCATCCGGATAGTT4651GTCTAAAAGGGAGAAAGGGATACAGTACTACAAAAGGAAAGCTATATTCG4701TACTTTGAGGGTACTAAATTTCACCAAGCAGCCAAAGATATGGCTGAAAT4751AAAAGTGCTGTTTCCGGACGACCAGGAAAGCAACGAACAGTTATGCGCTT4801ACATACTGGGCGAAACCATGGAAGCAATTCGTGAAAAATGCCCAGTTGAT4851CGTAACCCGTCATCCAGTCCTCCGAAGACGCTGCCTTGCCTTTGCATGTA4901TGCAATGACTCCGGAAAGAGTTCATAGGCTCAGAAGTAACAATGTTAAAG4951AAATTACTGTGTGCTCCTCGACCCCGCTTCCAAAATATAAGATTAAGAAC5001GTCCAGAAAGTCCAGTGCACTAAAGTAGTCCTGTTTAACCCGCACACTCC5051TACTTTTGTCCCGGCCCGTAAATATGTGGAAGTGCCAGAATCACCTGCCA5101TCACACCTGTACAGGCCGACACGCTAGATCAGCCACCTGCCGCGGACGGT5151ATACCGCTTGATGTTACGGACATTTCATTAAATATGGAAGATAGTAGCGA5201AGGATTGTCCATTTTAGATTTCCACGGGTCAGAAAGTTCCATTTTTAGCA5251TGGATAGCTGGTCGTCAGGAACCAGTTCTTTGGGGCCAGAGGACAATAGA5301AGGCAAGTAGTGACAGTCGATGTCCACTCCACCCAAGAGGATACTCCCAT5351TCCTCCTCCAAGGTTAAAGAAACTGGCCCGGTTAGCGGCGGCGAAACAGA5401CCCCAGTAGCACTTACCGTATCGAATGATGTGGGCTCAATGGATGAGTCC5451CTCTGCCTTTCATTTGGCAGCGTATCCATGTCTTTTGGATCTTTTTCCGA5501CGGTGAGATCGATGAAATAAGTCGTATGAAGACTGAGTCAGAACCCGTTT5551TATTTGGAACTTTTGAACCTGGAGAAGTTAATTCCATTATATCGTCTCGA5601TCAGCCGTGTCTTTTCCACCGCTAAGGCAGAGACGTAGACGTAGGAACAA5651GCGGACTGAATACTGACTAACCGGGGTAGGTGGGTACATATTTTCGACGG5701ATACAGGGCCAGGACATCTGCAAAAGAAGTCTGTCTTGCAGAATCAATTT5751TCCGAACCGACCTTGGAGCGTAACGTGCTGGAAAAGATATACGCTCCGAC5801GCTTGATACGTCGAAAGAAGAACTACTCAAATTTAGATACCAAATGATGC5851CCACCGAAGCCAATAAGAGCAGGTACCAGTCCCGCAAAGTCGAAAATCAA5901AAAGCCGTCACCACTGAGCGTTTGCTTTCAGGGTTACGGCTATATACCTC5951GGCAACTGATCAGCCTGAATGTTATAAAATTACTTACCCGAAACCTTTGT6001ATTCCAGCAGTGTACCAGCAAGTTACTCCGACCCGAAGTTCGCAGTTGCC6051GTCTGCAATAACTACTTGCATGAAAACTACCCAACGGTAGCGTCCTACCA6101GATTACTGATGAATACGACGCTTACCTCGATATGGTGGATGGGACTGTCG6151CTTGCCTGGACACCGCAACATTCTGCCCGGCTAAACTCAGAAGTTATCCA6201AAGAGGCATGAGTATCGCGCACCGAATATCCGTAGCGCAGTCCCGTCTGC6251TATGCAGAACACGTTGCAAAACGTGCTCATCGCTGCAACCAAGAGGAACT6301GCAACGTTACACAAATGCGTGAACTACCAACGCTGGACTCAGCGACGTTT6351AACGTCGAGTGCTTTAAAAAATATGCGTGCAACAATGAATACTGGGAAGA6401GTTTGCCCGTAAACCAATTAGAATTACCACGGAGTTCGTTACTGCGTATG6451TAGCCAGACTGAAAGGACCAAAGGCAGCAGCCTTATTTGCAAAGACGTAC6501AATTTGGTCCCATTGCAAGAGGTACCCATGGATAGGTTTGTTATGGATAT6551GAAAAGAGATGTGAAAGTCACACCAGGTACCAAGCACACCGAAGAGAGAC6601CAAAGGTGCAGGTAATCCAAGCTGCAGAGCCACTAGCGACGGCATATTTG6651TGCGGAATCCATCGCGAGTTGGTGCGTAGACTTACGGCGGTGTTGCTACC6701TAACATTCATACACTTTTTGATATGTCAGCAGAGGACTTTGACGCTATTA6751TAGCCGAACACTTTAAACATGGCGATCCGGTGCTGGAGACAGATATTGCT6801TCATTCGATAAGAGCCAAGATGACGCTATGGCGTTGACTGGCCTGATGAT6851CTTGGAAGATCTAGGCGTGGATCACCTACTACTCGACTTAATTGAGTGTG6901CCTTTGGTGAAATATCGTCAACACACTTGCCCACGGGCACTCGTTTCAAA6951TTTGGTGCAATGATGAAATCCGGAATGTTTCTTACATTATTCGTTAACAC7001AGTTTTGAATGTTGTGATCGCCTGCAGAGTACTAGAGGAAAGGCTTAAGA7051TGTCGAGATGCGCGGCGTTTATCGGCGACGATAACATTATACACGGGGTA7101GTGTCTGATAAGGTGATGGCAGAGAGGTGCGCCACCTGGCTTAACATGGA7151AGTTAAGATTATCGACGCCATTATCGGTGAGAGACCACCGTATTTTTGCG7201GTGGATTTATTCTGCAAGACTCAGTCACTTCCACAGCGTGCCGCGTGGCC7251GACCCACTGAAAAGATTATTTAAGCTTGGTAAACCGCTACCAGCCGACGA7301CGAGCAAGACGAAGATAGAAGGCGAGCTTTATTGGACGAAACAAAGGCGT7351GGTTCAGAGTGGGCATTACTGATACTTTAGCAACTGCTGTAGCAACCCGG7401TACGAGGTAGATAATATCACACCTGTCTTATTGGCATTGAGAACGTTTGC7451CCAGAGCAAAAATTCGTTTCAGGCCATTAGAGGAGAAATAAAGCAACTCT7501ACGGTGGTCCTAAATAGTCAGCATAGCATATTTTATCTGACTAATACTGT7551AACACCCCTACTATGAATAGAGGATTCTTTAACATGCTTGGCCGCCGCCC7601CTTCCCGGCCCCCACTTCTATGTGGAGGCCACGTAGAAGGAGGCAGGCGG7651CCCCGATGCCCGCCCGCAACGGGTTGGCTTCCCAAATTCAGCAGCTTACC7701ACAGCAGTTAGCGCCTTGGTTATAGGACAGGCAGCTAGACCTCAACAGCC7751ACGTCCTCGCCCGCCACCCCGCCAAAAGAAGCAGCCGCCAAAGCAATTAC7801CAAAACCGAAGAAACCGAAAACACAGGATAAGAAGAAGCGACAGCCCGCG7851AAGCCGAAACCTGGAAAGAGGCAGCGCATGGCGCTTAAGCTAGAGGCCGA7901CAGACTGTTTGATGTTAAGAACGAGAATGGTGATGTCATCGGACATGCTC7951TGGCCATGGAAGGAAAAGTAATGAAACCACTCCACGTGAAAGGAACTATA8001GATCACCCAGTGCTGTCGAAGCTTAAATTCACCAAGTCGTCGGCATATGA8051CATGGAGTTTGCACAACTTCCTGTCAATATGCGAAGCGAAGCTTTCACGT8101ACACCAGCGAGCACCCAGAAGGATTCTATAACTGGCACCATGGCGCAGTG8151CAGTATAGTGGAGGTAGATTCACTATTCCGCGTGGAGTAGGAGGCAGAGG8201TGACAGCGGCCGCCCGATTATGGACAACGCAGGCCGCGTCGTCGCTATCG8251TCCTCGGAGGAGCTGATGAGGGAACACGTACTGCCCTCTCGGTAGTTACC8301TGGAACAGTAAAGGAAAAACGATTAAGACATCCCCTGAGGGAACTGAAGA8351GTGGTCCGCAGCACCGCTGGTTACGGCCATGTGCTTACTTGGAAATGTAA8401GCTTCCCTTGCAACCGTCCGCCTACTTGCTACACCCGCGAACCATCTAGA8451GCGTTAGATATCCTGGAAGAGAATGTGAACCATGAGGACTACGACACTTT8501GCTCGACGCTATACTGCGATGCGATTTTTCCGGTAGAAATAAAAGAAGTG8551TTACCGGCGACTTTACCTTGACCAGCCCGTACCTAGGTACATGTCCATAC8601TGTCACCATACCGAGCCGTGCTTTAGTCCGATAAAGATCGAGCAGGTCTG8651GGATGAAGCGGATGACACCACTATACGCATCCAGACCTCTGCCCAATTTG8701GGTACGATCAGAGTGGAGCGACAAGCGTAAATAAATACCGCTACATGTCG8751TTCGATCAGGATCATACTGTGAAAGAAGGCCAGATGGACGATATTAAGAT8801CAGTACTTCAGGACCGTGCAGAAGACTTGGCCATAAAGGTTACTTCCTCC8851TTGCGAAATGTCCGCCCGGGGACAGCGTGACGGTCAGTATAGTGAGCAGC8901AGTTCAACTACATCATGCACGTTGGCACGTAAGATAAAACCGAAGTTTGT8951GGGACGGGAACGGTATGATCTGCCCCCCGTTTACGGAAAGAACATCCCTT9001GCAGAATGTACGACCGTCTGAAAGAAACGTCTGCTGGCTATATTACTATG9051CACAGACCTGGCCCACATGCCTACACGTCCTATCTGGAGGAAGCGTCCGG9101AAAAATTTACGCTAAGCCACCATCCGGAAAGAATATTACCTACGAGTGTA9151AATGCGGAGACTATAAAACAGGAACGGTAAAAACCCGCACTGAAATTACT9201GGCTGCACTGCCATTAAGCAGTGTGTCGCTTATAAGAGCGACCAAACGAA9251GTGGGTCTTTAACTCACCCGACCTGATAAGGCATGCCGATCACGCTGCAC9301AAGGAAAGTTGCATTTACCTTTTAGATTGGTCCCGAGTTCCTGCAAGGTC9351CCAGTGGCTCACGCACCGAGTGTAGTGCACGGCTTCAAACACATTAGCCT9401CCAACTGGATACGGACCACCTGACACTGCTCACCACCAGGAGACTAGGAG9451CGAATCCCGAACCAACCTCCGAATGGATTATCGGTAAGACTGTTAGAAAT9501TTTTCTGTTGGACGGGACGGCTTGGAGTATACGTGGGGCAACCACGACCC9551GGTAAGAGTTTACGCTCAAGAATCAGCGCCTGGCGATCCCCATGGATGGC9601CTCATGAGATAATACAGCATTACTATCACCGCCATCCTGCGTATACCATC9651CTAACCGTTGTCTCAGCTGCCGTTGCAGTGCTTATCGGCTTGACCGTTGC9701AGCACTGTGTACGTGTAAAGCGCGACGTGAATGTTTAACACCGTATGCCC9751TAGCCCCGAATGCCGTTGTTCCGACTTCGATAGCGCTCCTATGTTGCATT9801AGATCGGCTAACGCTGAAACATTTTCTGAGACGATGAGCTACCTGTGGTC9851AAACAGTCAGCCGTTCTTTTGGGCTCAGTTGTGTATCCCCTTAGCCGCTG9901TTGTCATCCTTGTGCGTTACTGTTCATGTTGCTTGCCTTTTTTAGTGGTT9951GCCGGCGTCTACCTGGGGAAGGTAGACGCCTACGAACATGCGACCACCAT10001TCCAAATGTGCCGAAGATTCCGTATAAAGCACTTGTAGAACGGTCTGGCT10051ACGCCCCGCTTAACTTAGAGATCACCGTCGTGTCGTCGCAGGTTTTACCA10101TCTACTAATCAAGAGTATATCACCTGCAAGTTTACTACTGTAGTCCCCTC10151CCCGAAAGTCAAATGTTGCGGCTCCTTGGAATGTCAGCCCGCCGCTCACG10201CCGACTATAACTGCAAAGTTTTTGGAGGGGTTTACCCATTCATGTGGGGA10251GGAGCACAGTGTTTTTGCGACAGCGAGAACACCCAGATGAGCGAGGCGTA10301CGTCAAACTGTCAGCGGACTGCGTAACGGATTACGCACAGGCAGTTAAGG10351TGCATACTGCTGCAATGAAGGTAGGACTACGCATAGTGTATGGGAATACC10401ACCAGCTATTTAGATGTGTACGTGAATGGAGTCACGCCGGGGACATCCAA10451AGATCTAAAAGTCATAGCTGGGCCAGTTTCGTCCTCCTTTACGCCGTTTA10501ATCATAAGGTTGTCATCTACCGTGGTTTAGTCTATAATTATGACTTCCCT10551GAATACGGCGCAATGAAACCTGGAGTGTTCGGAGACATCCAAGCTACCTC10601CTTGACCAGTAGAGACCTTATTGCAAGCACGGACATTAGATTGCTTAAGC10651CTTCCGTTAAAAACGTGCACGTCCCGTACACGCAAGCTGCTTCAGGATTC10701GAGATGTGGAAGAACAACTCAGGTCGACCACTGCAGGAAACCGCTCCTTT10751TGGGTGTAAGATTGCAGTCAACCCGCTACGAGCGGTGGACTGTTCGTACG10801GTAATATTCCTATCTCCATCGATATCCCAAATGCCGCCTTCATTAGGATA10851TCGGATGCGCCGCTAGTTTCGACGGTTAAATGTGAGGTCAGCGAATGTAC10901CTATTCAGCTGATTTTGGTGGTATGGCAACCCTGCAGTATGTGTCGGACC10951GCGAAGGACAATGCCCCGTGCACTCTCACTCAAGCACAGCAACACTTCAG11001GAATCGACAGTGCATGTCCTGGAGAAAGGGGCGGTGACCGTGCACTTTAG11051TACCGCCAGCCCACAGGCTAATTTCATTATATCGCTGTGTGGCAAGAAAA11101CAACATGCAACGCGGAATGTAAGCCACCTGCGGACCACATTGTGAGTACA11151CCGCACAAAATCGATCAGGAGTTCCAAACCGCTATTTCAAAAACATCATG11201GAGCTGGTTACTGGCACTCTTTGGCGGCGCCTCGTCGCTATTAATTATAG11251GACTCATGATTTTTACTTGCAGCATGTTGCTGACAAGCACCCGGAGATGA11301CCGCTATGTCCCAATGACCCGACCAGCAAAACTCGACGTATTTCCGAGGA11351ACTGATGTGCATAATGCATCAGGCTGGTATATTAGATCCCCGCTCATCGC11401GGGCACTATAGCAACAATGAAACTCGATGTATTTCCGAGGATGCGCAGTG11451CATAATGCTGCGCCGTGTTGCCATATAATCATTTTATTAATCAACTATTT11501AGCAGACGCTAAAACTCAATGTATTTCTGAGGAAGCGTGGTGCATAATGC11551CATGCAGCGTCTGTATAATTTCTTTTATTATTCTTTTATCAATTAACCAA11601AATTTTGTTTTTAACATTTCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA11651AAAAACTCGAGCAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATT11701TTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCA11751CTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATA11801CATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCA11851ATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCC11901TTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAA11951ACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGG12001TTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCC12051CCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGC12101GCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCAT12151ACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGC12201ATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACC12251ATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACC12301GAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCC12351TTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGT12401GACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAAC12451TGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGG12501AGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGC12551TGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTAT12601CATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCT12651ACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCT12701GAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTA12751CTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGA12801TCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGT12851GAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATC12901TTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAA12951AACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACT13001CTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGT13051CCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCAC13101CGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGT13151GGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGA13201TAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCT13251TGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGA13301GAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAG13351CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACG13401CCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGT13451CGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAG13501CAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA13551TGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCC13601TTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGA13651GTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATTTTCTCCTTA13701CGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATC13751TGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGT13801GACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCC13851CTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCG13901TCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGC13951GCGAGGCAGCTGCGGTAAAGCTCATCAGCGTGGTCGTGAAGCGATTCACA14001GATGTCTGCCTGTTCATCCGCGTCCAGCTCGTTGAGTTTCTCCAGAAGCG14051TTAATGTCTGGCTTCTGATAAAGCGGGCCATGTTAAGGGCGGTTTTTTCC14101TGTTTGGTCACTGATGCCTCCGTGTAAGGGGGATTTCTGTTCATGGGGGT14151AATGATACCGATGAAACGAGAGAGGATGCTCACGATACGGGTTACTGATG14201ATGAACATGCCCGGTTACTGGAACGTTGTGAGGGTAAACAACTGGCGGTA14251TGGATGCGGCGGGACCAGAGAAAAATCACTCAGGGTCAATGCCAGCGCTT14301CGTTAATACAGATGTAGGTGTTCCACAGGGTAGCCAGCAGCATCCTGCGA14351TGCAGATCCGGAACATAATGGTGCAGGGCGCTGACTTCCGCGTTTCCAGA14401CTTTACGAAACACGGAAACCGAAGACCATTCATGTTGTTGCTCAGGTCGC14451AGACGTTTTGCAGCAGCAGTCGCTTCACGTTCGCTCGCGTATCGGTGATT14501CATTCTGCTAACCAGTAAGGCAACCCCGCCAGCCTAGCCGGGTCCTCAAC14551GACAGGAGCACGATCATGCGCACCCGTGGCCAGGACCCAACGCTGCCCGA14601GAGCTCGATTTAGGTGACACTATAG其中：1-11620病毒核苷酸序列11621-11655polyA序列11656-11661XhoI酶切位点11656-14606pBR骨架质粒序列14601-14606SacI酶切位点14607-14625SP6启动子序列