免疫A型呼吸道合胞病毒G75-225蛋白的小鼠对病毒感染的抗性

目的:评价A型呼吸道合胞病毒(RSV)G75-225蛋白(G151)与二硫键异构酶(DsbA)的重组蛋白抗原DsbA-G151的免疫活性。方法:采用PCR方法从A型RSVG蛋白扩增G151基因片段,插入表达载体pET-DsbA,经E.coli表达、亲和层析纯化制备DsbA-G151蛋白;将其免疫BALB/c小鼠后获得相应的抗血清,利用ELISA方法、保护性实验检测蛋白免疫活性。结果:构建了表达载体pET-DsbA-G151,表达、纯化获得了重组蛋白DsbA-G151。ELISA检测表明,DsbA-G151能在小鼠体内产生高滴度的特异性IgG;保护性实验显示该蛋白能有效保护BALB/c小鼠不被RSV感染。结论:经ELISA检测、保护性实验,表明DsbA-G151具有良好的免疫原性。

The Mice Immunized by Protein G75-225 of RSV-A Resistant to Infection

Objective:To evaluate the immunogenicity of recombinant protein antigens DsbA-G151.Methods:G151 gene encoding G75-225(G151)of G protein of respiratory syncytial virus(RSV)type A was amplified by PCR and inserted into the pET-DsbA vector to construct an expression vector pET-DsbA-G151.Then it was transformed into E.coli to express the recombinant protein.BALB/c mice were immunized with one of the purified recombinant protein antigens DsbA-G151 for production of anti-serum.The immunogenicity of recombinant protein antigens was identified by ELISA and protective experiments.Results:The expression vector was constructed and the purified recombinant protein antigens were abtained.ELISA assays identified its developed specific serum antibodies.The mice were protected by recombinant protein in protective experiment.Conclusions:Based on ELISA assays and protected expirments,good immunogenicity of DsbA-G151 was revealed.