微小隐孢子虫TSP3蛋白质的表达与功能

Expression and Function of Cryptosporidium parvum TSP3 Protein

通过对微小隐孢子虫TSP3基因克隆构建原核表达载体,纯化TSP3 His-tag重组蛋白质免疫新西兰家兔,间接ELISA方法检测特异性血清效价并纯化多克隆抗体。纯化TSP3 GST-tag重组蛋白质并与HCT-8细胞进行共孵育,通过Western blot与间接ELISA检测细胞黏附特性。结果表明:成功构建了微小隐孢子虫p ET-TSP3和p GEX-TSP3重组表达质粒,并用亲和层析法成功纯化出较高质量的重组蛋白质。Cp TSP3重组蛋白质具有良好免疫原性,并能与HCT-8细胞黏附。

By cloning Cryptosporidium parvum TSP3, prokaryotic expression vector was constructed, and to immunize New Zealand rabbits, TSP3-His tag of recombinant protein was purified. By indirect ELISA method, specific serum titers were detected and polyclonal antibodies were purified. The pu- rified TSP3-GST tag of recombinant protein was incubated with HCT-8 cells by Western blot and in- direct ELISA cell adhesion properties. This study successfully constructed Cryptosporidium parvum pET-TSP3 and pGEX-TSP3 recombinant plasmid, and by affinity chromatography successfully puri- fied recombinant proteins of high quality. CpTSP3 recombinant proteins are of good immunogenicity and can bind HCT-8.