【摘要】 为探索牛瑟氏泰勒虫的表面蛋白基因融合产物作为双价疫苗的可行性,以牛瑟氏泰勒虫基因组DNA为模板,通过重叠延伸拼接聚合酶链式反应(SOE-PCR)把P23和P33表面蛋白基因连接一起,2个基因之间插入一个linker(Gly4Ser)3,经EcoRⅠ和XhoⅠ双酶切,获得1151bp的双基因融合片段,克隆于表达质粒pGEX-4T-1中,构建了双基因重组表达载体pGEX-4T-P23-P33,转化大肠杆菌BL21,经IPTG诱导,表达出预期大小70.0ku的融合蛋白。Western blot检测结果显示,该蛋白与牛瑟氏泰勒虫抗血清呈阳性反应,表明融合蛋白具有反应原性,为进一步研究此融合蛋白作为疫苗候选成分提供了理论依据。更多还原

【Abstract】 To develop a bivalent vaccine of express surface protein of bovine Theileria sergenti, P23 and P33 surface protein genes were linked by SOE-PCR separated by a (Gly4Ser) 3 linker. The 1 157 bp fusion gene was cloned into pGEX-4T-1 plasmid and transformed into E.coli BL21 for expression. SDS-PAGE and Western blot analysis showed that the recombinant fusion protein had a molecular weight of 70 ku and could be recognized by positive serum of cow Theileria sergenti.更多还原