牦牛病毒性腹泻病毒E1基因的原核表达  

Prokaryotic expression of E1 genes of bovine viral diarrhea virus and detection of the expressed protein

作　　者：王盼盼[1] 刘亚刚[1] 孙凯[1] 王文伯[1] 胡炳峰[1] 于吉锋[2] 冯英阳[1] 

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041 [2]四川省畜牧科学研究院,四川成都610041

出　　处：《西南民族大学学报（自然科学版）》2010年第5期763-766,共4页Journal of Southwest Minzu University(Natural Science Edition)

基　　金：四川省科技厅攻关项目(04ZY029-006-04);国家民委项目(07XN03)

摘　　要：以牦牛BVDV基因组DNA为模板,运用聚合酶链反应成功扩增出牦牛BVDV E1基因,将其插入到pET-28a及pET-32a原核表达载体,构建重组表达质粒pET-28a-E1和pET-32a-E1并分别转化至Rosetta(DE3)和BL21(DE3)宿主菌中,IPTG诱导表达,经SDS-PAGE检测,pET-28a-E1和pET-32a-E1在宿主菌Rosetta(DE3)和BL21(DE3)中均表达出约40KU的融合蛋白,结果分析表明E1基因在两个宿主中都获得了高效表达,表达出的蛋白大小与预期结果相符.By means of the BVDV genome DNA of the wild yak as the template for PCR,the gene E1 is amplified and inserted into PET-28a and PET-32a vector.The reconstructed plasmid pET-28a-E1and pET-32a-E1 are transformed into Rosetta（DE3）and BL21（DE3）host bacterium respectively.According to IPTG induce expression,a 40KU fusion protein is expressed by pET-32a-E1 in the Rosetta（DE3）and BL21（DE3）host bacterium and a 20KU fusion protein is expressed by pET-28a-E1 and PET-32a-E1 through SDS-PAGE detection.The results indicate that protein E1 is high effectively expressed in the host bacterium,and the protein size is consistent with the anticipated consequence.

关 键 词：牦牛 牛病毒性腹泻病毒 E1基因 原核表达 

分 类 号：S858.23[农业科学—临床兽医学]