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

笔者以山羊痘病弱毒疫苗毒株TK基因内KpnI为插入位点，构建了表达绿色荧光蛋白和小反刍兽疫（PPR）H基因的重组山羊痘病毒通用转移载体，为今后研究羊痘病毒活载体疫苗奠定基础。

Construction and Identification of a Goat Pox Virus Transfer Vector to Express Peste des Petits Ruminants H gene

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.