淋球菌OmpA蛋白的克隆表达、保守性及抗原表位分析

Clone and expression of OmpA protein from Neisseria gonorrhoeae and analysis of its conservation and B cell epitopes

目的了解OmpA蛋白在不同型别淋球菌间的保守性,分析OmpA蛋白的B细胞抗原表位,并通过大肠杆菌表达系统获取其重组蛋白。方法从Genebank数据库中获取不同型别淋球菌的OmpA蛋白氨基酸序列,利用Clustal X 2.1软件对序列保守性进行分析。利用DNASTAR软件预测分析OmpA蛋白B细胞抗原表位。构建pp SUMO-omp A重组表达载体,转化至大肠杆菌BL21(DE3)菌株中表达OmpA重组蛋白,以SDS-PAGE检测表达的重组蛋白。结果 OmpA蛋白在不同型别淋球菌菌株间保守性高达99.6%,OmpA蛋白抗原表位可能位于28-37、78-86、95-102、147-154、159-165、201-208、213-218等氨基酸区段。SDS-PAGE检测大肠杆菌中表达出OmpA蛋白,主要以包涵体形式存在。结论成功构建pp SUMO-omp A重组表达载体,获得包涵体形式的OmpA蛋白,且OmpA蛋白可能是一种保守性较好的淋球菌蛋白疫苗候选分子。

Objective To understand the conservation of OmpA protein among different,and to analyze the B cell antigen epitope of OmpA protein,and expression OmpA protein in E.coli.Methods Amino acid sequences of OmpA protein in different types of Neisseria gonorrhoeae were obtained from the Genebank database,The sequence conservation of OmpA proteins were analyzed using Clustal X 2.1 software.B cell antigen epitope of OmpA protein were predictively analyzed by DNASTAR software.Recombinant expression vector pp SUMO-omp A was constructed and identified.OmpA protein was expressed in E.coli BL21（DE3） strains and recombinant proteins were analyzed by SDS-PAGE.Results The conservation of OmpA proteins among different types of Neisseria gonorrhoeae strain was up to 99.6%.The most possible epitopes of OmpA protein were located within or nearby its N-terminal NO.78-86,28-37,147-154,159-165,201-208,and 213-218.OmpA protein was successfully expressed in E.coli,and it mainly exists in the form of the inclusion body.Conclusion Recombinant expression vector pp SUMO-omp A was successfully constructed and inclusion bodies of OmpA protein were obtained.OmpA protein may be a good conservative vaccine candidate targets of Neisseria gonorrhoeae.