山羊痘病毒P32基因真核表达载体的构建及表达  

Construction and Expression of Recombinant Antigen P32 Gene of Goatpox Virus

作　　者：谢芳[1] 张强[1] 颜新敏[1] 吴国华[1] 李健[1] 朱海霞[1] 郭爱疆[1] 韩若婵[1] 施程洪[1] 鞠厚斌[1] 朱彩珠[1] 岳城[2] 才学鹏[1] 

机构地区：[1]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,兰州730046 [2]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《新疆农业大学学报》2008年第3期1-4,共4页Journal of Xinjiang Agricultural University

基　　金：国家基础平台建设项目(2005DKA21205-3)

摘　　要：从重组克隆载体pMD18-P32中扩增出山羊痘病毒P32基因,与毕赤酵母分泌性表达载体pPIC9K相连接,构建重组表达载体pPIC9K-P32。重组质粒pPIC9K-P32用SalⅠ线性化后,与毕赤酵母(Pichia pastoris)GS115混合后电转化,使重组表达载体与酵母染色体发生同源重组。采用G418抗性梯度筛选法得到高拷贝重组菌株,甲醇诱导目的基因表达。SDS-PAGE和Western-blotting分析结果表明,用酵母成功表达出了31 kD的重组蛋白,该蛋白具有生物学活性,能被山羊痘阳性血清识别。The structural protein P32 gene of the goat pox virus in the recombinant plasmid pMD18-P32 was subcloned into the Pichia pastoris expression vector pPIC9K. The resultant recombinant plasmid pPIC9K-P32 was transformed into P. pastoris GSl15 by electroporation. The multi-copy recombinant P. pastoris strains were screened by G418 and induced by methanol. The expressed product was analyzed by SDS-PAGE and Western-blotting tests. The results indicated that the recombinant protein was expressed in P. pastoris, and had biological activity,the protein could be recognized by anti-serum against goat pox virus.

关 键 词：山羊痘病毒 P32基因 毕赤酵母 诱导表达 

分 类 号：S851.654[农业科学—预防兽医学]