重组柯萨奇B3病毒VP1基因的穿梭表达载体的构建及鉴定  

Construction and identification of Recombinant Bacterium-yeast shuttle vector for Coxsackievirus B3 VP1 gene

作　　者：黄孝天[1] 陈洪[1] 吴赟[1] 陈宏新[1] 刘晶星[1] 

机构地区：[1]上海第二医科大学病原生物学教研室

出　　处：《中国人兽共患病杂志》2004年第5期375-378,共4页Chinese Journal of Zoonoses

摘　　要：目的　构建重组柯萨奇B3病毒VP1基因的细菌 -酵母菌穿梭表达载体pGBKT7-VP1。方法　用PCR从质粒pT7-CV3- 5 5中扩增柯萨奇B3病毒VP1基因 ,经NdeⅠ和PstⅠ双酶切后 ,定向插入细菌 -酵母菌穿梭表达载体 pG BKT7中完成质粒构建。用酶切和测序鉴定构建质粒。结果　限制性内切酶酶切和序列分析表明VP1巳成功插入pGBKT7载体的多克隆位点 ,克隆方向和阅读框与病毒VP1基因一致。结论　本研究成功构建VP1基因的细菌 -酵母菌穿梭表达载体 pGBKT7-VP1,为研究VP1蛋白与其宿主细胞之间的相互作用奠定了基础。To construct recombinant bacterium yeast shuttle vector of Coxsackievirus B3 VP1 gene CVB3 VP1 gene was amplified from plasmid pT7 CV3 5 5 by PCR and directionally inserted into bacterium yeast shuttle vector pGBKT7 Then the recombinant plasmid was identified by restriction endonucleases digestion and sequencing After restriction endonucleases treatment and sequencing,it was comfirmed that VP1 gene was successfully inserted into the multi cloning site of vector pGBKT7 in the right direction and the open reading frame of VP1 gene in the recombinant plasmid was identical with that of Coxsackievirus B3 The recombinant bacterium yeast shuttle vector pGBKT7 VP1 was successfully constructed and made it possible to study the interaction between CVB3 VP1 protein and proteins of its host cell

关 键 词：柯萨奇133病毒 VP1 基因克隆 

分 类 号：R373.23[医药卫生—病原生物学]