猪流产嗜性衣原体POMP90蛋白的原核表达及反应原性分析

Prokaryotic Expression and Immune Reactivity of Swine Chlamydophila abortus POMP90 Protein

设计特异性引物进行PCR扩增,获得猪流产嗜性衣原体CP/12株的pomp90基因,将其定向克隆到原核表达载体pGEX-KG中。阳性重组质粒转化原核表达宿主菌E.coliBL21,用IPTG进行诱导表达,获得高效表达的融合蛋白。采用Western-blot和间接ELISA检测表明纯化的重组蛋白具有良好的反应原性,初步选择其最佳包被浓度为1μg/mL。

The object of this study is to obtain the recombinant polymorphic outer membrane POMP 90 protein of swine Chlamydophila abortus （CA） for development of diagnostic ELISA method. pompgO gene of swine CA strain CP/12 was obtained by PCR amplification and cloned into the prokaryotic expressive vector pGEX-KG. The recombinant plasmid was then expressed in the E. coli BL21 induced by IPTG. The purified recombinant POMP90 protein showed good immune reactivity detected by Western-blot and indirect ELISA,and the optimal concentration of antigen for coating was 1μg/mL.