CAJ | 학술논문

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pET?TSP3和pGEX?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。 CpTSP3重组蛋白质具有良好免疫原性，并能与HCT?8细胞黏附。
통과대미소은포자충TSP3기인극륭구건원핵표체재체，순화TSP3 His?tag중조단백질면역신서란가토，간접ELISA방법검측특이성혈청효개병순화다극륭항체。순화TSP3 GST?tag중조단백질병여HCT?8세포진행공부육，통과Western blot여간접ELISA 검측세포점부특성。결과표명：성공구건료미소은포자충pET?TSP3화pGEX?TSP3중조표체질립，병용친화층석법성공순화출교고질량적중조단백질。 CpTSP3중조단백질구유량호면역원성，병능여HCT?8세포점부。
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.