一种禽流感病毒基因重配株D3/F-R2/6及其构建方法

摘要：

一种禽流感病毒基因重配株D3/F-R2/6及其构建方法，涉及应用反向遗传技术。将MPAIV鸡胚高度适应株A/Chicken/Shanghai/F/98(H9N2)的6个内部基因PB2、PB1、PA、NP、M、NS和HPAIV流行株A/Duck/Huadong/D3/00(H5N1)的NA基因和致弱的HA基因进行重配，构建成禽流感病毒基因重配株D3/F-R2/6。可用于制造禽流感疫苗，以避免采用人流感病毒的内部基因和AIV的表面基因重配成新的A型流感病毒制备禽流感疫苗时，对公共卫生构成威胁。

专利类型：发明专利

申请/专利号：CN200510040973.9

申请日期：2005-07-08

公开/公告号：CN1733912

公开/公告日：2006-02-15

主分类号：C12N7/01(2006.01)I

申请/专利权人:扬州大学

发明/设计人:刘秀梵;石火英;卢建红;张小荣

主申请人地址:225009江苏省扬州市大学南路88号

专利代理机构:扬州市锦江专利事务所

代理人:江 平

国别省市代码:江苏;32

权利要求：

1、一种禽流感病毒基因重配株D3/F-R2/6，其特征在于其保藏号为CGMCCNO：1386，基因组长度为13.5Kb，由H5N1亚型AIV的两个外部基因和MPAIV鸡胚高度适应株A/Chicken/Shanghai/F/98(H9N2)的6个内部基因PB2、PB1、PA、NP、M、NS构成；所述H5N1亚型AIV的两个外部基因分别是HPAIV流行株A/Duck/Huadong/D3/00(H5N1)的NA基因和致弱的HA基因；HPAIV流行株A/Duck/Huadong/D3/00(H5N1)的致弱的HA基因DNA序列为：atggagaaaatagtgcttcttcttgcaatagtcagtcttgttaaaagtgatcagatttgc60attggttaccatgcaaacaactcgacagagcaggttgacacaataatggaaaagaacgtt120actgttacgcatgcccaagacatactggaaaaggcacacaacgggaaactctgcgatcta180gatggagtgaagcctctaattttgggagattgtagtgtagctggatggctcctcggaaac240cctatgtgtgacgaattcatcaatgtgccggaatggtcttacatagtggggaaggccagt300ccagccaatgacctctgttacccaggggatttcaacgactatgaagaactgaaacaccta360ttgagcagaataaaccactttgagaaaattcagatcatccccaaaagttcttggtccaat420catgaagcctcatcaggggtgagcgcggcatgtccataccatgggaagccctcctttttc480agaaatgtggtatggcttatcaaaaagaacagtgcatacccaacaataaagaggagctac540aataacaccaaccaagaagatcttttggtactgtgggggattcaccatcctaatgatgca600gcagagcagacaaagctctatcaaaacccaaccacctatatctccgttggaacatcaaca660ttaaaccagagattggtcccaaaaatagctactagatccagagtaaacgggcaaagtgga720agaatggagttcttctggacaattttaaagccgaatgatgccataaatttcgagagtaat780gggaatttcattgctccagaatacgcatacaaaattgtcaagaaaggggactcagcaatt840atgaaaagtgaattggaatatggtaactgcaacaccaagtgtcaaactccaatgggggcg900ataaactctagtatgccattccacaacatacaccctctcaccatcggggaatgccccaaa960tatgtgaaatcaaacagattagtccttgcgactagactcagaaatacccctcaaagagag1020agaaggagaaaaaagagaggactatttggagctatagcaggttttatagagggaggatgg1080cagggaatggtagatggttggtatgggtaccaccatagcaatgagcaggggagtggatac1140gctgcagacaaagaatccactcaaaaggcaatagatggagtcaccaataaggtcaactcg1200atcattgacaaaatgaacactcagtttgaggccgttggaagggaatttaataacttagaa1260aggaggatagaaaatttaaacaagaagatggaagacggattcctagatgtctggacttat1320aatgctgaacttctggttctcatggaaaatgagagaactctagactttcatgattcaaat1380gtcaagaacctttacaacaaggtccgactacagcttagggataatgcaaaggagctgggt1440aatggttgtttcgagttctatcacaaatgtgataatgaatgtatggaaagtgtaaaaaac1500gggacgtatgactacccgcagtattcagaagaagcaagactaaacagagaggaaataaat1560ggagtaaaattggaatcaatgggaacttaccaaatactgtcaatttattcaacagtggcg1620agttccctagcactggcaatcatggtagctggtctatctttatggatgtgctccaatgga1680tcgttacaatgcagaatttgcatttaaHPAIV流行株A/Duck/Huadong/D3/00(H5N1)的NA基因DNA序列为：agcaaaagcaggagttcaaaatgaatccaaatcaaaagataataaccaccggatcaatct60gtatggtaattggaatggttagcttgatgttgcaaattgggaacataatctcaatatggg120ttagtcattcaattcagacagggactcaacaccgagctgaaccatgcaatcgaagcatta180ttaattatgaaaacaacacctgggtaaatcagacatatgtcaaaatcagcaacaccaatt240ttcttactgagaaagctgtggcttcagtaagattagtgggcaattcatctctttgcccca300tgtttgttataagagagccgttcatctcatgctcccacttggaatgccgaaccttctttt360tgactcagggagccttgctgaatgataagcactccaatgggaccgtcaaagacagaagcc420cttacagaacattgatgagctgtcctgtgggtgaggccccctccccatataactcgaggt480ttgagtctgttgcttggtcggcaagtgcttgtcatgatggcactagttggttgacaattg540gaatttctggcccagacaatggggctgtggctgtattgaaatacaatggcataataacag600acactatcaagagttggaggaacaacatactgagagctcaagagtctgaatgtgcatgtg660taaatggctcttgctttactgtaatgactgacggaccaagtaatgggcaggcttcatata720agatcttcaaaatagaaaaagggaaagtagttaaatcagtcgaattgaatgcccctaatt780atcactatgaggagtgctcctgttatcctgatgctggcgaaatcacatgtgtgtgcaggg840ataattggcatggctcaaatcggccatgggtatctttcaatcaaaatttggagtaccaaa900taggatatatatgcagtggagttttcggagacaatccacgccccaatgatggaacaggca960gttgtggtccggtgtcccctaacggggcatatggagtaaaagggttttcatttaaatacg1020gcaatggtgtttggatcgggagaaccaaaagcactaattccaggagcggctttgaaatga1080tttgggatccaaatgggtggactggaacggacagtaatttttcggtgaagcaagatatcg1140tagctataactgattggtcaggatatagcgggagttttgtccagcatccagaactgacag1200gattagattgcataagaccttgtttctgggttgagctaatcagagggcggcccaaagaga1260gcacaatttggactagtgggagcagcatatctttttgtggtgtaaatagtgacactgtgg1320gttggtcttggccagacggtgctgagttgccattcaccattgacaagtagtttgttcaaa1380aaactccttgtttctact。