CAJ | 학술논문

在成功克隆牛病毒性腹泻一黏膜病痛毒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鏊定.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.
재성공극륭우병독성복사일점막병통독Changchun184주E2기인적기출상,장E2기인여표체재체pMV261련접,구건료중조천사질립pMV261-E2,후전전화도잡개묘중,획득료구유잡나매소항성적중조잡개묘,병대중조잡개묘진행균락PCR오정.재45℃하열유도E2기인재중조잡개묘중적표체,병대표체산물진행SDS-PAGE전영화Wesern blotting 분석.결과증명,우병독성복사-점막병병독Changchun184주E2기인재잡개묘중성공적표체.
E2 gene of BVDV Changchun 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 kanamyein.The recombinant BCG were identifieated by PCR.E2 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.