| 牛瑟氏泰勒虫MPSP双表位基因的克隆及原核表达 |
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| 引用本文： | 钱年超,贾立军,薛书江,张影,张守发.牛瑟氏泰勒虫MPSP双表位基因的克隆及原核表达[J].中国预防兽医学报,2011,33(10). |
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| 作者姓名： | 钱年超 贾立军 薛书江 张影 张守发 |
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| 作者单位： | 延边大学农学院/动物医学系,吉林延吉,133002 |
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| 基金项目： | 国家自然科学基金(30960278) |
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| 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 (Major piroplasm 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BL21,IPTG诱导表达,经SDS-PAGE电泳显示表达了约39 ku的融合蛋白.Westem 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.
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| 关 键 词： | 牛瑟氏泰勒虫 MPSP 表位 原核表达 |
Prokaryotic expression of MPSP double-epitope gene of bovine Theileria sergenti |
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| QIAN Nian-chao,JIA Li-jun,XUE Shu-jiang,ZHANG Ying,ZHANG Shou-fa.Prokaryotic expression of MPSP double-epitope gene of bovine Theileria sergenti[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(10). |
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| Authors: | QIAN Nian-chao JIA Li-jun XUE Shu-jiang ZHANG Ying ZHANG Shou-fa |
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| Institution: | QIAN Nian-chao,JIA Li-jun,XUE Shu-jiang,ZHANG Ying,ZHANG Shou-fa (Department of Veterinary Medicine,College of Agricultural,Yanbian University,Yanji 133002,China) |
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| Abstract: | 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 BL2l inducted by IPTG,SDS-PAGE analysis showed that recom... |
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| Keywords: | Theileria sergenti MPSP epitope prokaryotic expression |
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