牛瑟氏泰勒虫HSP70基因的克隆及原核表达  

Cloning and procaryotic expression of HSP70 gene of bovine Theileria sergenti

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

机构地区：[1]延边大学农学院动物医学系,吉林龙井133400

出　　处：《黑龙江畜牧兽医》2010年第6期16-18,共3页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(30560112)

摘　　要：根据GenBank(D12692.1)上发表的牛瑟氏泰勒虫HSP70基因序列设计、合成1对特异性的引物,采用PCR方法扩增牛瑟氏泰勒虫HSP70基因片段,将扩增产物与pMD18-Tsimple载体连接,重组质粒经PCR、酶切鉴定后测序;用pGEX-4T-1表达载体构建重组质粒pGEX-HSP70,经IPTG诱导表达后进行SDS-PAGE、Western-blot分析。结果表明:扩增的HSP70基因与D12692.1序列同源性为99.3%;表达的融合蛋白分子质量约为68ku,而且可与牛瑟氏泰勒虫阳性血清发生特异性反应。A pair of specific primers were designed and synthetized according to the HSP70 gene sequence（D12692.1）of bovine Theileria sergenti which was published in GenBank.HSP70 gene fragment of bovine Theileria sergenti was amplified by PCR and ligated into pMD18-T simple vector.The recombinant plasmid was sequenced after being certified by PCR amplification and restriction analysis.The pGEX-HSP70 was constructed using pGEX-4T-1 and analyzed by SDS-PAGE and Western-blot after it was induced and expressed by IPTG.The result showed that the sequence homology between HSP70 gene and D12692.1 was 99.3%,the molecular weight of the expressed fusion protein was about 68 ku,the expressed fusion protein could be recognized by the positive serum of bovine Theileria sergenti.

关 键 词：牛瑟氏泰勒虫 HSP70 克隆 原核表达 

分 类 号：S852.7[农业科学—基础兽医学]