淋球菌膜蛋白疫苗的研究

Study on the vaccine of membrane protein of gonorrhoeae

目的 :设计淋球菌膜蛋白疫苗并构建原核表达系统 ,测定表达蛋白的免疫活性。方法 :PCR扩增淋球菌膜蛋白( PIA)基因片段 ,构建重组质粒 ,重组质粒宿主菌经 IPTG诱导 ,用 SDS- PAGE分析 PIA的表达情况。淋球菌标准株免疫 BAL B/c小鼠 ,体外凝集检测动物血清的免疫效果及 EL ISA法检测 PIA的免疫活性。结果 :成功构建表达 PIA蛋白的重组质粒 ,SDS-PAGE电泳图上显示 1条相对分子量 ( Mr)约为 4 0 KDa的新生条带 ,能与免疫血清发生特异性结合反应。结论 :淋球菌膜蛋白疫苗原核表达系统的设计和构建正确 ,表达蛋白 PIA具有免疫活性 。

Objective:To design the vaccine of membrane protein of gonorrhoeae, construct the system of prokaryotic expression, detect the immune activity of the expressive protein.Methods:The DNA fragment of membrane protein(PIA) of gonorrhoeae was amplified by PCR and recombinant plasmids of the DNA was constructed. After introduction of the plasmids into host bacterium, the expressed proteins of the plasmids, upon IPTG induction, was examined by SDS-PAGE. BALB/C mice were immunized with standard strain of gonorrhoeae. The immunization effect of animal serum was detected in vitro by agglutination reaction, and the antigenicity of PIA was proved by indirect ELISA.Results:The recombinant plasmids expressing PIA protein were successfully constructed. The SDS-PAGE showed a new lane of 40 kDa in molecular weight.The latter can specifically combine with immune serum.Conclusion:The design and construction of the vaccine of membrane protein of gonorrhoeae in E.coli is correct. The expressed protein, PIA, possesses immune activity.