牛病毒性腹泻-黏膜病病毒Changchun184株E_2基因在卡介苗中的表达

Expressing of bovine viral diarrhea-mucosal disease virus Changchun184 strain of E_2 gene in BCG

在成功克隆牛病毒性腹泻-黏膜病病毒Changchun184株E2基因的基础上,将E2基因与表达载体pMV261连接,构建了重组穿梭质粒pMV261-E2,后电转化到卡介苗中,获得了具有卡那霉素抗性的重组卡介苗,并对重组卡介苗进行菌落PCR鉴定。在45℃下热诱导E2基因在重组卡介苗中的表达,并对表达产物进行SDS-PAGE电泳和Wesern blotting分析。结果证明,牛病毒性腹泻-黏膜病病毒Changchun184株E2基因在卡介苗中成功的表达。

E2 gene of BVDV Changehun 184 strain was cloned and inserted into the shuttle expression plasmid vector pMV261,the recombinant shuttle plasmid pMV261-E2 was constructed. Then pMV261-E2 was transformed into BCG successfully and obtained recombinant BCG which was resistive to kanamycin, The recombinant BCG were identificated by PCR. Ez gene expression in recombinant BCG was induced in 45℃, then the SDS-PAGE and western blotting was used to analyze the expression product. The results indicated the BVDV E2 gene was expressd in BCG successfully.