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采用PCR构建编码B群脑膜炎球菌fHBP蛋白V1变异型全长序列和V2变异型C结构域抗原表位融合蛋白的基因片段。并克隆入pET30a(+)载体,转化大肠杆菌BL21(DE3),在其实现表达,表达量约占菌体蛋白总量的30%。表达产物经离子交换层析纯化后,获得了纯度达到90%以上的融合蛋白。将融合蛋白免疫小鼠,对其免疫原性进行了初步分析。结果显示,经2次腹腔免疫,血清IgG滴度达到15 849,同时杀菌力实验显示此融合蛋白能诱导针对B群脑膜炎球菌的补体依赖的杀菌反应。表明此融合蛋白具有较好的免疫原性,为B群脑膜炎球菌蛋白疫苗的研究提供了一定的理论基础。
PCR was used to construct the gene fragment encoding the full-length sequence of variant fV1 of fHBP protein of group B meningococcal and V2 variant C-domain epitope fusion protein. And cloned into pET30a (+) vector, which was then transformed into E. coli BL21 (DE3) for expression in about 30% of the total bacterial proteins. After purified by ion-exchange chromatography, the fusion protein with the purity of more than 90% was obtained. The fusion protein was immunized in mice, and its immunogenicity was analyzed preliminarily. The results showed that after 2 times of intraperitoneal immunization, serum IgG titer reached 15 849, and bactericidal test showed that the fusion protein could induce complement-dependent bactericidal response against meningococcus group B. The results showed that the fusion protein had good immunogenicity and provided a theoretical basis for the study of serogroup B meningococcal protein vaccine.