牛瑟氏泰勒虫MPSP双表位基因的克隆及原核表达  

Prokaryotic expression of MPSP double-epitope gene of bovine Theileria sergenti

作　　者：钱年超[1] 贾立军[1] 薛书江[1] 张影[1] 张守发[1] 

机构地区：[1]延边大学农学院/动物医学系,吉林延吉133002

出　　处：《中国预防兽医学报》2011年第10期790-793,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30960278)

摘　　要：为了探究表位疫苗是否可以应用于预防牛瑟氏泰勒虫(Theileria sergenti),根据MPSP(Majorpiroplasm surface protein)基因序列(FJ515689.1)设计合成了2对引物,以T.sergenti基因组DNA为模板,通过SOE-PCR扩增出长约361 bp的双表位基因融合片段,2个表位基因通过linker(Gly4Ser)3相连。将该片段连接pGEX-4T-2,构建pGEX-4T-E1-2重组表达载体,转化BL2l,IPTG诱导表达,经SDS-PAGE电泳显示表达了约39 ku的融合蛋白。Western blot分析表明该双表位重组蛋白可与T.sergenti阳性血清发生特异性反应,表明融合蛋白具有反应原性。To express two major piroplasm surface protein （MPSP） epitopes of Theileria sergenti, two pairs of primers were designed and synthesized according to the gene sequence of T. sergenti in GenBank. T. sergenti genomic DNA was used as the template and 361 bp double-epitope fusion gene was amplified by SOE-PCR which was connected by linker（Gly4Ser）3. The fragment was cloned into pGEX-4T-2 to construct pGEX-4T-E1-2. The recombinant protein was expressed in E. coli BL21 inducted by IPTG, SDS-PAGE analysis showed that recombinant protein was about 39 ku, which was recognized by positive serum of T. sergenti.

关 键 词：牛瑟氏泰勒虫 MPSP 表位 原核表达 

分 类 号：S852.7[农业科学—基础兽医学]