[发明专利]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。

主权项

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： 1 ATTGACGGCG TAGTACACAC TATTGAATCA AACAGCCGAC CAATAGCACT 51 ACCATCATAA TGGAGAGGCC TGTAGTTAAC GTAGACGTAG ACCCCCAGAG 101 TCCGTTTGTC GCTCAACTGC AAAAGAGCTT CCCGCAATTC GAGGTAGTAG 151 CACAGCAGGC CACACCAAAT GACCATGCTA ATGCCAGAGC ATTTTCGCAT 201 CTGGCTAGTA AATTAATCGA GCTGGAGGTT CCTACCACAG CGACGATTTT 251 GGACATAGGC AGCGCACCGG CTCGTAGAAT GTTTTCCGAG CACCACTATC 301 ACTGCGTCTG CCCTATGCGG AGCCCCGAAG ATCCCGACCG TATGATGAAA 351 TACGCCAATA AGTTGGCGGA GAAGGCAAAT AAGATTACTA ATAAAAATTT 401 GCATGAGAAG ATTAAAGACC TCCGGATCGT ACTCGATACT CCGGATGCTG 451 AGACACCGTC GCTCTGCTTC CATAATGACG TTACCTGCAG TACGCGTGCA 501 GAGTACTCCG TTATGCAAGA TGTGTATATT AATGCACCCG GAACTATTTA 551 CCATCAAGCT ATGAAAGGCG TGCGGACACT TTACTGGATT GGGTTTGACA 601 CCACTCAGTT CATGTTCTCG GCTATGGCAG GATCATATCC TGCCTATAAT 651 ACTAACTGGG CCGATGAAAA AGTCCTTGAA GCACGCAATA TTGGACTCTG 701 TAGTACCAAG TTGAGCGAAG GCCGTATAGG AAAGTTGTCA ATAATGAGGA 751 AAAAGGAGTT GAAGCCCGGG TCACGGGTCT ACTTCTCAGT GGGATCAACA 801 CTCTACCCAG AATATAGGGA CAGCTTACAG AGCTGGCACC TCCCATCAGT 851 GTTCCATCTG AAAGGAAAGC AATCGTATAC ATGCCGCTGT GATACAGTGG 901 TAAGTTGCGA AGGCTACGTA GTGAAGAAGA TCACTATTAG TCCCGGGATT 951 ACGGGAGAAA CCGTGGGATA CGCGGTTACA AACAATAGCG AGGGTTTCTT1001 GCTATGTAAA GTTACTGACA CAGTAAAAGG GGAACGGGTC TCGTTCCCCG1051 TGTGCACGTA TATCCCGGCC ACCATATGCG ACCAGATGAC AGGTATAATG1101 GCCACGGATA TTTCACCTGA CGATGCACAG AAGCTTCTGG TTGGGCTCAA1151 CCAGAGGATT GTCATTAACG GTAAAACCAA CAGGAACACT AACACCATGC1201 AAAACTACCT TCTACCGATT ATAGCACAAG GCTTCAGCAA GTGGGCTAAA1251 GAGCGCAAAG AAGATCTTGA TAATGAAAAG AAGCTGGGCA CTAGGGAGCG1301 TAAGCTTATT TACGGGTGTC TATGGGCGGT TCGTACTAAG AAAGTGCACT1351 CGTTTTACCG CCCGCCCGGA ACGCAGACCA GCGTGAAAGT CCCGGCATCT1401 TTTAGCGCTT TTCCAATGTC ATCTGTATGG ACAACCTCAC TACCCATGTC1451 GCTGAGGCAG AAGATAAAAT TGGTACTACA ACCGAAAAAG GAGGAGAAAT1501 TACTGCAGGT CTCAGAAGAG TTAGTTGCGG AGGCTAAAGC GGCCTTTGAA1551 GATGCACAGG AGGAGATCAG AGCGGAGCAA CTCCGTGAAG CACTTCCACC1601 ACTGGTAGCA GACAAAGGTA TTGAAGCCGC TGCAGAAGTC GTCTGTGAAG1651 TGGAAGGGCT TCAAGCTGAT ATAGGAGCAG CTCTTGTTGA GACGCCACGA1701 GGGCATGTAA GGATTATACC TCAAGTAACA GACCGCATGA TTGGGCAGTA1751 CATCGTGGTC TCACCAACCT CCGTGCTTAA GAACGCCAAA TTAACACCTG1801 TTCACCCTTT GGCTGACCAA GTCAAAATTA TAACACATTC AGGGAGGACA1851 GGAAGATTCG CGGTGGAGCC GTATGATGCT AAAGTGTTGA TGCCAGCAGG1901 TAGCGCCGTT CCATGGCCTG AGTTCCTGGC GTTAAGTGAA AGCGCCACGC1951 TAGTGTATAA CGAAAGAGAA TTTGTCAACC GCAAACTTTA CCATATCGCC2001 ATACATGGTC CCGCGAAGAA TACTGAAGAG GAGCAGTATA AAGTCACTAA2051 AGCCGAACTC GCAGAAACAG AGTATGTATT TGATGTCGAC AAGAAGCGTT2101 GCGTTAAGAA GGAAGAGGCC TCGGGGCTGG TTCTCTCGGG AGAACTAACT2151 AACCCACCGT ATCATGAAAT GGCGCTTGAG GGGCTGAAGA CTCGACCCGC2201 TGTTCCGTAT AAGGTCGAAA CAATAGGAGT AATAGGCACA CCAGGATCAG2251 GCAAGTCTGC GATTATTAAA TCGACCGTTA CCGTACGAGA TCTTGTTACC2301 AGCGGAAAGA AGGAAAACTG CCGTGAAATT GAAACTGACG TGTTGAGGTT2351 GAGAGGTATG CAGATCACGT CAAGGACGGT AGACTCAGTC ATGCTTAACG2401 GATGCCATAA AGCCGTTGAG GTGCTATACG TTGATGAAGC ATTTGCGTGC2451 CATGCTGGTA CGTTGCTTGC CTTGATCGCT ATTGTTAGAC CCCGAAAGAA2501 GGTAGTACTT CGCGGGGATC CTAAGCAGTG TGGTTTTTTC AATATGATGC2551 AGCTCAAAGT ACATTTTAAT CACCCTGAAA AAGATATATG TACTAAGACG2601 TTTTACAAAT TCATCTCTCG ACGCTGCACG CAACCAGTAA CGGCGATTGT2651 TTCAACACTT CATTACGACG GAAAGATGAA GACTACAAAC CCCTGCAAGA2701 AGAGCATTGA GATAGATATT ACAGGTACTA CGAAGCCGAA GCCCGGGGAC2751 CTCGTTTTGA CGTGCTTCCG CGGATGGGTC AAGCAGCTAC AGATCGATTA2801 CGCGGGAAAT GAAGTGATGA CGGCTGCTGC CTCGCAGGGA CTGACTAGAA2851 AGGGTGTTTA CGCCGTTCGG CAAAAAGTTA ATGAGAATCC ACTGTACGCG2901 ATTACGTCGG AGCACGTGAA CGTGTTACTC ACCCGTACTG AGGATAGATT2951 AGTGTGGAAG ACCTTACAGG GCGATCCATG GATCAAACAA CTTACTAACA3001 TTCCGAAAGG AAACTTCCAA GCTACTATTG AGGACTGGGA AGCTGAACAT3051 AAGGGGATCA TCGCTGCAAT AAACAGCCCA ACCCCTCGTA TAAACCCGTT3101 CAGCTGTAAG ACTAATGTGT GCTGGGCGAA GGCACTGGAA CCGATACTGG3151 CCACAGCCGG TATTGTCCTC ACCGGTTGCC AGTGGAGTGA GCTGTTTCCA3201 CAGTTCGTAG ATGATAAACC CCACTCAGCT ATCTACGCCC TAGATGTGAT3251 TTGTATTAAG TTCTTTGGTA TGGATCTGAC AAGCGGTCTA TTTTCAAAGC3301 AGAGCATCCC TCTGACGTAC CACCCTGCGG ACTCTGCAAG GCCAGTGGCG3351 CATTGGGACA ACAGTCCAGG AACCCGAAAG TATGGATACG ATCATGCGGT3401 TGCTGCCGAA TTGTCTCGTA GATTCCCGGT GTTCCAACTT GCTGGAAAAG3451 GCACACAGCT TGACTTGCAG ACTGGTAGAA CCAGAGTCGT TTCCGCGCAG3501 TGTAACTTGG TCCCAGTGAA CCGTAACCTC CCGCACGCTC TTGTCCCCGA3551 GTATAAAGAG AAACAACCCG GCCCGATTAA AAATTTTTTA AATCAGTTTA3601 AGCATCACTC CATACTTGTA GTATCAGAAA CGAAAATCGA AGTTCCCAAT3651 AAGCGCATCG AATGGATTGC ACCGCTTGGC ATAGCTGGTG CAGATAAGAG3701 CTACAACCTG GCCTTCGGAT TTCCACCGCA GGCACGGTAT GATATGGTGT3751 TTATCAATAT AGGAACAAAA TATAGAAACC ACCATTTTCA ACAGTGTGAA3801 GACCATGCGG CGACTTTGAA GACTCTTTCC CGCTCGGCTC TGAATTGCCT3851 CAACCCTGGA GGCACCTTAG TGGTGAAATC CTATGGATAT GCTGATCGCA3901 ATAGCGAGGA CGTAGTCACC GCACTTGCCA GGAAGTTTGT TAGAGTGTCT3951 GCGGCCAGGC CAGAGTGCGT CTCAAGTAAC ACAGAAATGT ACCTAATCTT4001 TCGGCAATTA GATAATAGCC GTACACGGCA GTTCACTCCA CATCATTTGA4051 ACTGTGTAAT CTCGTCGGTG TATGAAGGCA CGAGAGAAGG AGTCGGAGCT4101 GCGCCATCCT ACCGTGTGAA ACGGGAAAAT ATTGCAGACT GCCATGAGGA4151 AGCAATCGTC AACGCCGCTA ACTCGCTGGG TAAACCAGGT GAAGGAGTTT4201 GCCGCGCCGT CTACAAGCGT TGGCCGAGCA GCTTTATGGA TTCCGCCACA4251 GAAACGGGTA CGGCTAAATT AACTGTAAGC CAAGGAATGA AAGTGATACA4301 CGCGGTCGGC CCTGACTTCC GTAAGTATCC TGAGGCGGAA GCTTTGAAGC4351 TGCTGCAAAA CGCTTACCAT GCAGTGGCGG ACTTGGTTAA CAAACACAAC4401 ATTAAGTCCA TTGCTATCCC GCTACTATCA ACAGGTATAT ATGCAGCTGG4451 TAAGGATCGC TTGGAAGTTT CGCTCAATTG CCTGACCACC GCACTAGACA4501 GGACCGATGC AGATGTAACT ATTTATTGTT TGGATAAGAA ATGGAAAGAG4551 AGAATTGACG CGGTGTTGCA ACTTAAGGAG TCAGTGACAG AACTGAAGGA4601 CGAGGACATG GAAATCGATG ACGAATTGGT ATGGATTCAT CCGGATAGTT4651 GTCTAAAAGG GAGAAAGGGA TACAGTACTA CAAAAGGAAA GCTATATTCG4701 TACTTTGAGG GTACTAAATT TCACCAAGCA GCCAAAGATA TGGCTGAAAT4751 AAAAGTGCTG TTTCCGGACG ACCAGGAAAG CAACGAACAG TTATGCGCTT4801 ACATACTGGG CGAAACCATG GAAGCAATTC GTGAAAAATG CCCAGTTGAT4851 CGTAACCCGT CATCCAGTCC TCCGAAGACG CTGCCTTGCC TTTGCATGTA4901 TGCAATGACT CCGGAAAGAG TTCATAGGCT CAGAAGTAAC AATGTTAAAG4951 AAATTACTGT GTGCTCCTCG ACCCCGCTTC CAAAATATAA GATTAAGAAC5001 GTCCAGAAAG TCCAGTGCAC TAAAGTAGTC CTGTTTAACC CGCACACTCC5051 TACTTTTGTC CCGGCCCGTA AATATGTGGA AGTGCCAGAA TCACCTGCCA5101 TCACACCTGT ACAGGCCGAC ACGCTAGATC AGCCACCTGC CGCGGACGGT5151 ATACCGCTTG ATGTTACGGA CATTTCATTA AATATGGAAG ATAGTAGCGA5201 AGGATTGTCC ATTTTAGATT TCCACGGGTC AGAAAGTTCC ATTTTTAGCA5251 TGGATAGCTG GTCGTCAGGA ACCAGTTCTT TGGGGCCAGA GGACAATAGA5301 AGGCAAGTAG TGACAGTCGA TGTCCACTCC ACCCAAGAGG ATACTCCCAT5351 TCCTCCTCCA AGGTTAAAGA AACTGGCCCG GTTAGCGGCG GCGAAACAGA5401 CCCCAGTAGC ACTTACCGTA TCGAATGATG TGGGCTCAAT GGATGAGTCC5451 CTCTGCCTTT CATTTGGCAG CGTATCCATG TCTTTTGGAT CTTTTTCCGA5501 CGGTGAGATC GATGAAATAA GTCGTATGAA GACTGAGTCA GAACCCGTTT5551 TATTTGGAAC TTTTGAACCT GGAGAAGTTA ATTCCATTAT ATCGTCTCGA5601 TCAGCCGTGT CTTTTCCACC GCTAAGGCAG AGACGTAGAC GTAGGAACAA5651 GCGGACTGAA TACTGACTAA CCGGGGTAGG TGGGTACATA TTTTCGACGG5701 ATACAGGGCC AGGACATCTG CAAAAGAAGT CTGTCTTGCA GAATCAATTT5751 TCCGAACCGA CCTTGGAGCG TAACGTGCTG GAAAAGATAT ACGCTCCGAC5801 GCTTGATACG TCGAAAGAAG AACTACTCAA ATTTAGATAC CAAATGATGC5851 CCACCGAAGC CAATAAGAGC AGGTACCAGT CCCGCAAAGT CGAAAATCAA5901 AAAGCCGTCA CCACTGAGCG TTTGCTTTCA GGGTTACGGC TATATACCTC5951 GGCAACTGAT CAGCCTGAAT GTTATAAAAT TACTTACCCG AAACCTTTGT6001 ATTCCAGCAG TGTACCAGCA AGTTACTCCG ACCCGAAGTT CGCAGTTGCC6051 GTCTGCAATA ACTACTTGCA TGAAAACTAC CCAACGGTAG CGTCCTACCA6101 GATTACTGAT GAATACGACG CTTACCTCGA TATGGTGGAT GGGACTGTCG6151 CTTGCCTGGA CACCGCAACA TTCTGCCCGG CTAAACTCAG AAGTTATCCA6201 AAGAGGCATG AGTATCGCGC ACCGAATATC CGTAGCGCAG TCCCGTCTGC6251 TATGCAGAAC ACGTTGCAAA ACGTGCTCAT CGCTGCAACC AAGAGGAACT6301 GCAACGTTAC ACAAATGCGT GAACTACCAA CGCTGGACTC AGCGACGTTT6351 AACGTCGAGT GCTTTAAAAA ATATGCGTGC AACAATGAAT ACTGGGAAGA6401 GTTTGCCCGT AAACCAATTA GAATTACCAC GGAGTTCGTT ACTGCGTATG6451 TAGCCAGACT GAAAGGACCA AAGGCAGCAG CCTTATTTGC AAAGACGTAC6501 AATTTGGTCC CATTGCAAGA GGTACCCATG GATAGGTTTG TTATGGATAT6551 GAAAAGAGAT GTGAAAGTCA CACCAGGTAC CAAGCACACC GAAGAGAGAC6601 CAAAGGTGCA GGTAATCCAA GCTGCAGAGC CACTAGCGAC GGCATATTTG6651 TGCGGAATCC ATCGCGAGTT GGTGCGTAGA CTTACGGCGG TGTTGCTACC6701 TAACATTCAT ACACTTTTTG ATATGTCAGC AGAGGACTTT GACGCTATTA6751 TAGCCGAACA CTTTAAACAT GGCGATCCGG TGCTGGAGAC AGATATTGCT6801 TCATTCGATA AGAGCCAAGA TGACGCTATG GCGTTGACTG GCCTGATGAT6851 CTTGGAAGAT CTAGGCGTGG ATCACCTACT ACTCGACTTA ATTGAGTGTG6901 CCTTTGGTGA AATATCGTCA ACACACTTGC CCACGGGCAC TCGTTTCAAA6951 TTTGGTGCAA TGATGAAATC CGGAATGTTT CTTACATTAT TCGTTAACAC7001 AGTTTTGAAT GTTGTGATCG CCTGCAGAGT ACTAGAGGAA AGGCTTAAGA7051 TGTCGAGATG CGCGGCGTTT ATCGGCGACG ATAACATTAT ACACGGGGTA7101 GTGTCTGATA AGGTGATGGC AGAGAGGTGC GCCACCTGGC TTAACATGGA7151 AGTTAAGATT ATCGACGCCA TTATCGGTGA GAGACCACCG TATTTTTGCG7201 GTGGATTTAT TCTGCAAGAC TCAGTCACTT CCACAGCGTG CCGCGTGGCC7251 GACCCACTGA AAAGATTATT TAAGCTTGGT AAACCGCTAC CAGCCGACGA7301 CGAGCAAGAC GAAGATAGAA GGCGAGCTTT ATTGGACGAA ACAAAGGCGT7351 GGTTCAGAGT GGGCATTACT GATACTTTAG CAACTGCTGT AGCAACCCGG7401 TACGAGGTAG ATAATATCAC ACCTGTCTTA TTGGCATTGA GAACGTTTGC7451 CCAGAGCAAA AATTCGTTTC AGGCCATTAG AGGAGAAATA AAGCAACTCT7501 ACGGTGGTCC TAAATAGTCA GCATAGCATA TTTTATCTGA CTAATACTGT7551 AACACCCCTA CTATGAATAG AGGATTCTTT AACATGCTTG GCCGCCGCCC7601 CTTCCCGGCC CCCACTTCTA TGTGGAGGCC ACGTAGAAGG AGGCAGGCGG7651 CCCCGATGCC CGCCCGCAAC GGGTTGGCTT CCCAAATTCA GCAGCTTACC7701 ACAGCAGTTA GCGCCTTGGT TATAGGACAG GCAGCTAGAC CTCAACAGCC7751 ACGTCCTCGC CCGCCACCCC GCCAAAAGAA GCAGCCGCCA AAGCAATTAC7801 CAAAACCGAA GAAACCGAAA ACACAGGATA AGAAGAAGCG ACAGCCCGCG7851 AAGCCGAAAC CTGGAAAGAG GCAGCGCATG GCGCTTAAGC TAGAGGCCGA7901 CAGACTGTTT GATGTTAAGA ACGAGAATGG TGATGTCATC GGACATGCTC7951 TGGCCATGGA AGGAAAAGTA ATGAAACCAC TCCACGTGAA AGGAACTATA8001 GATCACCCAG TGCTGTCGAA GCTTAAATTC ACCAAGTCGT CGGCATATGA8051 CATGGAGTTT GCACAACTTC CTGTCAATAT GCGAAGCGAA GCTTTCACGT8101 ACACCAGCGA GCACCCAGAA GGATTCTATA ACTGGCACCA TGGCGCAGTG8151 CAGTATAGTG GAGGTAGATT CACTATTCCG CGTGGAGTAG GAGGCAGAGG8201 TGACAGCGGC CGCCCGATTA TGGACAACGC AGGCCGCGTC GTCGCTATCG8251 TCCTCGGAGG AGCTGATGAG GGAACACGTA CTGCCCTCTC GGTAGTTACC8301 TGGAACAGTA AAGGAAAAAC GATTAAGACA TCCCCTGAGG GAACTGAAGA8351 GTGGTCCGCA GCACCGCTGG TTACGGCCAT GTGCTTACTT GGAAATGTAA8401 GCTTCCCTTG CAACCGTCCG CCTACTTGCT ACACCCGCGA ACCATCTAGA8451 GCGTTAGATA TCCTGGAAGA GAATGTGAAC CATGAGGACT ACGACACTTT8501 GCTCGACGCT ATACTGCGAT GCGATTTTTC CGGTAGAAAT AAAAGAAGTG 8551 TTACCGGCGA CTTTACCTTG ACCAGCCCGT ACCTAGGTAC ATGTCCATAC 8601 TGTCACCATA CCGAGCCGTG CTTTAGTCCG ATAAAGATCG AGCAGGTCTG 8651 GGATGAAGCG GATGACACCA CTATACGCAT CCAGACCTCT GCCCAATTTG 8701 GGTACGATCA GAGTGGAGCG ACAAGCGTAA ATAAATACCG CTACATGTCG 8751 TTCGATCAGG ATCATACTGT GAAAGAAGGC CAGATGGACG ATATTAAGAT 8801 CAGTACTTCA GGACCGTGCA GAAGACTTGG CCATAAAGGT TACTTCCTCC 8851 TTGCGAAATG TCCGCCCGGG GACAGCGTGA CGGTCAGTAT AGTGAGCAGC 8901 AGTTCAACTA CATCATGCAC GTTGGCACGT AAGATAAAAC CGAAGTTTGT 8951 GGGACGGGAA CGGTATGATC TGCCCCCCGT TTACGGAAAG AACATCCCTT 9001 GCAGAATGTA CGACCGTCTG AAAGAAACGT CTGCTGGCTA TATTACTATG 9051 CACAGACCTG GCCCACATGC CTACACGTCC TATCTGGAGG AAGCGTCCGG 9101 AAAAATTTAC GCTAAGCCAC CATCCGGAAA GAATATTACC TACGAGTGTA 9151 AATGCGGAGA CTATAAAACA GGAACGGTAA AAACCCGCAC TGAAATTACT 9201 GGCTGCACTG CCATTAAGCA GTGTGTCGCT TATAAGAGCG ACCAAACGAA 9251 GTGGGTCTTT AACTCACCCG ACCTGATAAG GCATGCCGAT CACGCTGCAC 9301 AAGGAAAGTT GCATTTACCT TTTAGATTGG TCCCGAGTTC CTGCAAGGTC 9351 CCAGTGGCTC ACGCACCGAG TGTAGTGCAC GGCTTCAAAC ACATTAGCCT 9401 CCAACTGGAT ACGGACCACC TGACACTGCT CACCACCAGG AGACTAGGAG 9451 CGAATCCCGA ACCAACCTCC GAATGGATTA TCGGTAAGAC TGTTAGAAAT 9501 TTTTCTGTTG GACGGGACGG CTTGGAGTAT ACGTGGGGCA ACCACGACCC 9551 GGTAAGAGTT TACGCTCAAG AATCAGCGCC TGGCGATCCC CATGGATGGC 9601 CTCATGAGAT AATACAGCAT TACTATCACC GCCATCCTGC GTATACCATC 9651 CTAACCGTTG TCTCAGCTGC CGTTGCAGTG CTTATCGGCT TGACCGTTGC 9701 AGCACTGTGT ACGTGTAAAG CGCGACGTGA ATGTTTAACA CCGTATGCCC 9751 TAGCCCCGAA TGCCGTTGTT CCGACTTCGA TAGCGCTCCT ATGTTGCATT 9801 AGATCGGCTA ACGCTGAAAC ATTTTCTGAG ACGATGAGCT ACCTGTGGTC 9851 AAACAGTCAG CCGTTCTTTT GGGCTCAGTT GTGTATCCCC TTAGCCGCTG 9901 TTGTCATCCT TGTGCGTTAC TGTTCATGTT GCTTGCCTTT TTTAGTGGTT 9951 GCCGGCGTCT ACCTGGGGAA GGTAGACGCC TACGAACATG CGACCACCAT10001 TCCAAATGTG CCGAAGATTC CGTATAAAGC ACTTGTAGAA CGGTCTGGCT10051 ACGCCCCGCT TAACTTAGAG ATCACCGTCG TGTCGTCGCA GGTTTTACCA10101 TCTACTAATC AAGAGTATAT CACCTGCAAG TTTACTACTG TAGTCCCCTC10151 CCCGAAAGTC AAATGTTGCG GCTCCTTGGA ATGTCAGCCC GCCGCTCACG10201 CCGACTATAA CTGCAAAGTT TTTGGAGGGG TTTACCCATT CATGTGGGGA10251 GGAGCACAGT GTTTTTGCGA CAGCGAGAAC ACCCAGATGA GCGAGGCGTA10301 CGTCAAACTG TCAGCGGACT GCGTAACGGA TTACGCACAG GCAGTTAAGG10351 TGCATACTGC TGCAATGAAG GTAGGACTAC GCATAGTGTA TGGGAATACC10401 ACCAGCTATT TAGATGTGTA CGTGAATGGA GTCACGCCGG GGACATCCAA10451 AGATCTAAAA GTCATAGCTG GGCCAGTTTC GTCCTCCTTT ACGCCGTTTA10501 ATCATAAGGT TGTCATCTAC CGTGGTTTAG TCTATAATTA TGACTTCCCT10551 GAATACGGCG CAATGAAACC TGGAGTGTTC GGAGACATCC AAGCTACCTC10601 CTTGACCAGT AGAGACCTTA TTGCAAGCAC GGACATTAGA TTGCTTAAGC10651 CTTCCGTTAA AAACGTGCAC GTCCCGTACA CGCAAGCTGC TTCAGGATTC10701 GAGATGTGGA AGAACAACTC AGGTCGACCA CTGCAGGAAA CCGCTCCTTT10751 TGGGTGTAAG ATTGCAGTCA ACCCGCTACG AGCGGTGGAC TGTTCGTACG10801 GTAATATTCC TATCTCCATC GATATCCCAA ATGCCGCCTT CATTAGGATA10851 TCGGATGCGC CGCTAGTTTC GACGGTTAAA TGTGAGGTCA GCGAATGTAC10901 CTATTCAGCT GATTTTGGTG GTATGGCAAC CCTGCAGTAT GTGTCGGACC10951 GCGAAGGACA ATGCCCCGTG CACTCTCACT CAAGCACAGC AACACTTCAG11001 GAATCGACAG TGCATGTCCT GGAGAAAGGG GCGGTGACCG TGCACTTTAG11051 TACCGCCAGC CCACAGGCTA ATTTCATTAT ATCGCTGTGT GGCAAGAAAA11101 CAACATGCAA CGCGGAATGT AAGCCACCTG CGGACCACAT TGTGAGTACA11151 CCGCACAAAA TCGATCAGGA GTTCCAAACC GCTATTTCAA AAACATCATG11201 GAGCTGGTTA CTGGCACTCT TTGGCGGCGC CTCGTCGCTA TTAATTATAG11251 GACTCATGAT TTTTACTTGC AGCATGTTGC TGACAAGCAC CCGGAGATGA11301 CCGCTATGTC CCAATGACCC GACCAGCAAA ACTCGACGTA TTTCCGAGGA11351 ACTGATGTGC ATAATGCATC AGGCTGGTAT ATTAGATCCC CGCTCATCGC11401 GGGCACTATA GCAACAATGA AACTCGATGT ATTTCCGAGG ATGCGCAGTG11451 CATAATGCTG CGCCGTGTTG CCATATAATC ATTTTATTAA TCAACTATTT11501 AGCAGACGCT AAAACTCAAT GTATTTCTGA GGAAGCGTGG TGCATAATGC11551 CATGCAGCGT CTGTATAATT TCTTTTATTA TTCTTTTATC AATTAACCAA11601 AATTTTGTTT TTAACATTTC AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA11651 AAAAACTCGA GCAATTCTTG AAGACGAAAG GGCCTCGTGA TACGCCTATT11701 TTTATAGGTT AATGTCATGA TAATAATGGT TTCTTAGACG TCAGGTGGCA11751 CTTTTCGGGG AAATGTGCGC GGAACCCCTA TTTGTTTATT TTTCTAAATA11801 CATTCAAATA TGTATCCGCT CATGAGACAA TAACCCTGAT AAATGCTTCA11851 ATAATATTGA AAAAGGAAGA GTATGAGTAT TCAACATTTC CGTGTCGCCC11901 TTATTCCCTT TTTTGCGGCA TTTTGCCTTC CTGTTTTTGC TCACCCAGAA11951 ACGCTGGTGA AAGTAAAAGA TGCTGAAGAT CAGTTGGGTG CACGAGTGGG12001 TTACATCGAA CTGGATCTCA ACAGCGGTAA GATCCTTGAG AGTTTTCGCC12051 CCGAAGAACG TTTTCCAATG ATGAGCACTT TTAAAGTTCT GCTATGTGGC12101 GCGGTATTAT CCCGTGTTGA CGCCGGGCAA GAGCAACTCG GTCGCCGCAT12151 ACACTATTCT CAGAATGACT TGGTTGAGTA CTCACCAGTC ACAGAAAAGC12201 ATCTTACGGA TGGCATGACA GTAAGAGAAT TATGCAGTGC TGCCATAACC12251 ATGAGTGATA ACACTGCGGC CAACTTACTT CTGACAACGA TCGGAGGACC12301 GAAGGAGCTA ACCGCTTTTT TGCACAACAT GGGGGATCAT GTAACTCGCC12351 TTGATCGTTG GGAACCGGAG CTGAATGAAG CCATACCAAA CGACGAGCGT12401 GACACCACGA TGCCTGCAGC AATGGCAACA ACGTTGCGCA AACTATTAAC12451 TGGCGAACTA CTTACTCTAG CTTCCCGGCA ACAATTAATA GACTGGATGG12501 AGGCGGATAA AGTTGCAGGA CCACTTCTGC GCTCGGCCCT TCCGGCTGGC12551 TGGTTTATTG CTGATAAATC TGGAGCCGGT GAGCGTGGGT CTCGCGGTAT12601 CATTGCAGCA CTGGGGCCAG ATGGTAAGCC CTCCCGTATC GTAGTTATCT12651 ACACGACGGG GAGTCAGGCA ACTATGGATG AACGAAATAG ACAGATCGCT12701 GAGATAGGTG CCTCACTGAT TAAGCATTGG TAACTGTCAG ACCAAGTTTA12751 CTCATATATA CTTTAGATTG ATTTAAAACT TCATTTTTAA TTTAAAAGGA12801 TCTAGGTGAA GATCCTTTTT GATAATCTCA TGACCAAAAT CCCTTAACGT12851 GAGTTTTCGT TCCACTGAGC GTCAGACCCC GTAGAAAAGA TCAAAGGATC12901 TTCTTGAGAT CCTTTTTTTC TGCGCGTAAT CTGCTGCTTG CAAACAAAAA12951 AACCACCGCT ACCAGCGGTG GTTTGTTTGC CGGATCAAGA GCTACCAACT13001 CTTTTTCCGA AGGTAACTGG CTTCAGCAGA GCGCAGATAC CAAATACTGT13051 CCTTCTAGTG TAGCCGTAGT TAGGCCACCA CTTCAAGAAC TCTGTAGCAC13101 CGCCTACATA CCTCGCTCTG CTAATCCTGT TACCAGTGGC TGCTGCCAGT13151 GGCGATAAGT CGTGTCTTAC CGGGTTGGAC TCAAGACGAT AGTTACCGGA13201 TAAGGCGCAG CGGTCGGGCT GAACGGGGGG TTCGTGCACA CAGCCCAGCT13251 TGGAGCGAAC GACCTACACC GAACTGAGAT ACCTACAGCG TGAGCTATGA13301 GAAAGCGCCA CGCTTCCCGA AGGGAGAAAG GCGGACAGGT ATCCGGTAAG13351 CGGCAGGGTC GGAACAGGAG AGCGCACGAG GGAGCTTCCA GGGGGAAACG13401 CCTGGTATCT TTATAGTCCT GTCGGGTTTC GCCACCTCTG ACTTGAGCGT13451 CGATTTTTGT GATGCTCGTC AGGGGGGCGG AGCCTATGGA AAAACGCCAG13501 CAACGCGGCC TTTTTACGGT TCCTGGCCTT TTGCTGGCCT TTTGCTCACA13551 TGTTCTTTCC TGCGTTATCC CCTGATTCTG TGGATAACCG TATTACCGCC13601 TTTGAGTGAG CTGATACCGC TCGCCGCAGC CGAACGACCG AGCGCAGCGA13651 GTCAGTGAGC GAGGAAGCGG AAGAGCGCCT GATGCGGTAT TTTCTCCTTA13701 CGCATCTGTG CGGTATTTCA CACCGCATAT GGTGCACTCT CAGTACAATC13751 TGCTCTGATG CCGCATAGTT AAGCCAGTAT ACACTCCGCT ATCGCTACGT13801 GACTGGGTCA TGGCTGCGCC CCGACACCCG CCAACACCCG CTGACGCGCC13851 CTGACGGGCT TGTCTGCTCC CGGCATCCGC TTACAGACAA GCTGTGACCG13901 TCTCCGGGAG CTGCATGTGT CAGAGGTTTT CACCGTCATC ACCGAAACGC13951 GCGAGGCAGC TGCGGTAAAG CTCATCAGCG TGGTCGTGAA GCGATTCACA14001 GATGTCTGCC TGTTCATCCG CGTCCAGCTC GTTGAGTTTC TCCAGAAGCG14051 TTAATGTCTG GCTTCTGATA AAGCGGGCCA TGTTAAGGGC GGTTTTTTCC14101 TGTTTGGTCA CTGATGCCTC CGTGTAAGGG GGATTTCTGT TCATGGGGGT14151 AATGATACCG ATGAAACGAG AGAGGATGCT CACGATACGG GTTACTGATG14201 ATGAACATGC CCGGTTACTG GAACGTTGTG AGGGTAAACA ACTGGCGGTA14251 TGGATGCGGC GGGACCAGAG AAAAATCACT CAGGGTCAAT GCCAGCGCTT14301 CGTTAATACA GATGTAGGTG TTCCACAGGG TAGCCAGCAG CATCCTGCGA14351 TGCAGATCCG GAACATAATG GTGCAGGGCG CTGACTTCCG CGTTTCCAGA14401 CTTTACGAAA CACGGAAACC GAAGACCATT CATGTTGTTG CTCAGGTCGC14451 AGACGTTTTG CAGCAGCAGT CGCTTCACGT TCGCTCGCGT ATCGGTGATT14501 CATTCTGCTA ACCAGTAAGG CAACCCCGCC AGCCTAGCCG GGTCCTCAAC14551 GACAGGAGCA CGATCATGCG CACCCGTGGC CAGGACCCAA CGCTGCCCGA14601 GAGCTCGATT TAGGTGACAC TATAG其中：1-11620 病毒核苷酸序列 11621-11655 polyA序列 11656-11661 XhoI酶切位点 11656-14606 pBR骨架质粒序列 14601-14606 SacI酶切位点 14607-14625 SP6启动子序列