伪狂犬病病毒Bartha株gC膜外区部分基因的原核表达及纯化  

Prokaryotic Expression and Purification of the Extracellular Domain gC Gene of Pseudorabies Virus Bartha Strain

作　　者：刘庆伟[1,2] 邱亚峰[1] 王晓杜[1] 郭林[1] 刘超[1] 沈阳[1] 李向东[1] 单虎[2] 马志永[1] 

机构地区：[1]中国农业科学院上海兽医研究所兽医公共卫生实验室,上海200241 [2]青岛农业大学动物科技学院,山东青岛266109

出　　处：《动物医学进展》2009年第2期1-4,共4页Progress In Veterinary Medicine

基　　金：国家自然科学基金青年基金(30700593);中央级公益性科研院所基本科研业务费专项资金项目

摘　　要：构建伪狂犬病病毒(PRV)gC膜外区部分基因的重组质粒,原核表达并纯化目的蛋白。根据GenBank已发表的伪狂犬病病毒Bartha(PRV Ba)株gC基因的序列(NC EU719641),设计并合成了1对引物,以PRV Ba株为模板,PCR扩增出PRV gC膜外区部分基因片段;将该基因片段克隆到原核表达载体pET-28a上,转化大肠埃希菌BL21(DE3),经IPTG诱导,获得大小为35 ku的重组蛋白,命名为pET-gCN813。按照His-Bind纯化试剂盒说明书纯化表达产物,获得融合蛋白的纯化产物。To construct the recombinant plasmid of the extracellular domain gC gene of PRV, prokaryotic expression of the gene and purification of the fusion protein were conducted. A pair of primers were designed based on the gC gene sequence of Pseudorabies virus Bartha strain （PRV Ba） published in GenBank （NC EU719641）. The partial extracellular domain gC gene of PRV Ba was amplified by PCR, and cloned into pET-28a to construct the recombinant expression plasmid. Then it was transformed into E. coli BL21 （DE3）. A 35 ku fusion protein, named pET gCN813, was expressed in high yeild after IPTG induction. Whereafter, the fusion protein was successfully purified by His Bind Kit.

关 键 词：伪狂犬病病毒 GC基因 原核表达 融合蛋白 纯化 

分 类 号：S852.659.1[农业科学—基础兽医学]