安徽农业科学2009，Vol.37Issue(20)：9384-9386,9404,4.
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
Construction and Identification of a Goat pox Virus Transfer Vector to Express Peste des Petits Ruminants Hgene
邵长春 1高世功 2张强 3吴国华 2颜新敏 2李健 2王建科 2卢晓丽 2赵志荀 2崔丽凡2
作者信息
- 1. 甘肃农业大学,甘肃兰州,730070
- 2. 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,甘肃兰州,730046
- 3. 中农威特生物科技股份有限公司
- 折叠
摘要
Abstract
[Objective]This study was to develop a live vector goat pox virus vaccine 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 KpnⅠ, 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 after transfection. [Conclusion] The construction of goat poxvirus live vector laid a foundation for the live vector vaccine of PPR vaccine.关键词
羊痘病毒/H基因/转移载体/构建/鉴定Key words
Goat pox virus/ H gene/ Transfer vector/ Construction/ Identification分类
农业科技引用本文复制引用
邵长春,高世功,张强,吴国华,颜新敏,李健,王建科,卢晓丽,赵志荀,崔丽凡..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[J].安徽农业科学,2009,37(20):9384-9386,9404,4.
