| 表达小反刍兽疫病毒H基因的山羊痘病毒通用转移载体的构建及鉴定(英文) |
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| 引用本文： | 邵长春,张强,吴国华,颜新敏,李健,王建科,卢晓丽,赵志荀,崔丽凡,高世功.表达小反刍兽疫病毒H基因的山羊痘病毒通用转移载体的构建及鉴定(英文)[J].Agricultural Science&Technology,2009(3). |
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| 作者姓名： | 邵长春 张强 吴国华 颜新敏 李健 王建科 卢晓丽 赵志荀 崔丽凡 高世功 |
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| 作者单位： | 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室;农业部畜禽病毒学重点开放实验室; |
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| 摘 要： | Objective] This study was to develop a live vector vaccine of goat pox virus of Peste des petits ruminants(PPR). Method] Using PCR amplification technique, PPR H gene was obtained, then ligated into pGEM-T easy vector; the recombinants were digested by Nhe Ⅰ and Hind Ⅲ, and ligated into pEGFP-N1-P7.5, yielding the recombinant vector pEGFP-N1-P7.5-H; next the expression cassette EGFP-N1-P7.5-H was first released from recombinant vector pEGFP-N1-P7.5-H by double digestion of Hind Ⅲ and Nhe Ⅰ and ligated into pUC119-TK that was digested by Kpn Ⅰ, yielding the transfer vector pUC119-TK-EGFP-P7.5-H. Result] Identification and double enzyme digestion showed that the transfer vector pUC119-TK-EGFP-P7.5-H was correctly constructed. From the transfer vector transfected BHK-21 cells which infected GTPV AV41, specific fluorescence was observed at 48th h of transfection. Conclusion] The construction of goat poxvirus live vector laid a foundation for the live vector vaccine of PPR vaccine.
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| 关 键 词： | 表达 通用转移载体 绿色荧光蛋白 酶切位点 小反刍兽疫 重组基因 山羊痘病毒 重组质粒 酶切鉴定 病毒感染 |
Construction and Identification of a Goat Pox Virus Transfer Vector to Express Peste des Petits Ruminants H gene |
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| SHAO Chang-chun,ZHANG Qiang,WU Guo-hua,YAN Xin-min,LI Jian,WANG Jian-ke,LU Xiao-li,ZHAO Zhi-xun,CUI Li-fan,GAO Shi-gong State Key Lab of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences,Key Laboratory of Animal Virology of Ministry of Agriculture,Lanzhou.Construction and Identification of a Goat Pox Virus Transfer Vector to Express Peste des Petits Ruminants H gene[J].农业科学与技术(英文版),2009(3). |
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| Authors: | SHAO Chang-chun ZHANG Qiang WU Guo-hua YAN Xin-min LI Jian WANG Jian-ke LU Xiao-li ZHAO Zhi-xun CUI Li-fan GAO Shi-gong State Key Lab of Veterinary Etiological Biology Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences Key Laboratory of Animal Virology of Ministry of Agriculture Lanzhou |
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| Institution: | SHAO Chang-chun,ZHANG Qiang,WU Guo-hua,YAN Xin-min,LI Jian,WANG Jian-ke,LU Xiao-li,ZHAO Zhi-xun,CUI Li-fan,GAO Shi-gong State Key Lab of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences,Key Laboratory of Animal Virology of Ministry of Agriculture,Lanzhou 730046 |
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| Abstract: | Objective] This study was to develop a live vector vaccine of goat pox virus of Peste des petits ruminants(PPR). Method] Using PCR amplification technique, PPR H gene was obtained, then ligated into pGEM-T easy vector; the recombinants were digested by Nhe Ⅰ and Hind Ⅲ, and ligated into pEGFP-N1-P7.5, yielding the recombinant vector pEGFP-N1-P7.5-H; next the expression cassette EGFP-N1-P7.5-H was first released from recombinant vector pEGFP-N1-P7.5-H by double digestion of Hind Ⅲ and Nhe Ⅰ and ligated into pUC119-TK that was digested by Kpn Ⅰ, yielding the transfer vector pUC119-TK-EGFP-P7.5-H. Result] Identification and double enzyme digestion showed that the transfer vector pUC119-TK-EGFP-P7.5-H was correctly constructed. From the transfer vector transfected BHK-21 cells which infected GTPV AV41, specific fluorescence was observed at 48th h of transfection. Conclusion] The construction of goat poxvirus live vector laid a foundation for the live vector vaccine of PPR vaccine. |
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| Keywords: | Goat pox virus H gene Transfer vector Construction Identification |
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