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)

作者：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

来源：Agricultural Science & Technology, 2009, 10(03): 15-18+35.

DOI：10.16175/j.cnki.1009-4229.2009.03.033

• Goat pox virus
• H gene
• Transfer vector
• Construction
• Identification

[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.

• 单位
  中国农业科学院兰州兽医研究所; 农业部; 家畜疫病病原生物学国家重点实验室