肺炎链球菌TCSs中WalR的克隆表达、保守性及抗原表位分析

发布时间：2019-05-18 23:25

【摘要】：通过构建PET28a-WalR表达载体并在大肠杆菌BL21中表达WalR重组蛋白,以评价其对C57小鼠的免疫原性;并分析其在不同血清型肺炎链球菌中的保守性和B细胞的抗原表位。以肺炎链球菌D39血清型WalR基因为模板,构建重组质粒PET28a-WalR并转入BL21诱导表达目的蛋白质。采用Clustal X 2.1软件对WalR蛋白在不同血清型S.pn中的保守性进行分析;利用DNASTAR软件对其B细胞抗原表位进行分析。结果显示,成功构建PET28a-Wal R重组表达载体,经IPTG诱导后有大量高纯度的目的蛋白质,但其主要以包涵体形式存在;Wal R蛋白在不同血清型肺炎链球菌中高达99.4%,其B细胞抗原表位可能位于9-16、35-42、95-111、113-135、150-161、200-218等氨基酸序列位点。成功获得大量以包涵体形式表达的肺炎链球菌Wal R重组蛋白,并对其保守性和抗原表位进行了系统分析。
[Abstract]:PET28a-WalR expression vector was constructed and WalR recombinant protein was expressed in E. coli BL21 to evaluate its immunogenicity to C57 mice, and its conservatism in different serotypes of Streptococcus pneumoniae and antigen epitopes of B cells were analyzed. Using Streptococcus pneumoniae D39 serotype WalR as template, the recombinant plasmid PET28a-WalR was constructed and transferred into BL21 to induce the expression of the target protein. The conservatism of WalR protein in different serotypes of S.pn was analyzed by Clustal X 2.1 software, and its B cell epitope was analyzed by DNASTAR software. The results showed that PET28a-Wal R recombinant expression vector was successfully constructed and a large number of high purity target proteins were induced by IPTG, but they mainly existed in the form of inclusion bodies. Wal R protein is as high as 99.4% in different serotypes of Streptococcus pneumoniae. Its B cell epitope may be located at 9: 16, 35 / 42, 95: 111113: 135150 / 161200 / 218 and other amino acid sites. A large number of recombinant proteins of Streptococcus pneumoniae Wal R expressed in the form of inclusion bodies were successfully obtained and their conserved and antigenic epitopes were systematically analyzed.
【作者单位】：遵义医学院医学检验系;遵义医学院附属医院检验科;
【基金】：国家自然科学基金资助项目(81460317) 贵州省科技攻关项目(黔科合SY字(2012)3092号)
【分类号】：R446.1

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