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目的构建汉坦病毒SEO型代表株L99G2蛋白的多表位抗原基因(mea)。方法通过生物信息学软件对L99株G2蛋白氨基酸序列进行综合分析及预测,优选B细胞表位,引入GPG间隔序列串联表位,设计mea,然后应用重叠PCR法构建mea,并将其克隆到原核表达质粒pET32a(+)。结果优选出5个B细胞表位,设计并成功构建mea,PCR定向克隆获得pET32a-mea重组表达质粒。结论首次构建了汉坦病毒G2糖蛋白mea及其原核表达系统E.coliBL21/pET32a-mea,为其表达及免疫学应用奠定基础。
Objective To construct the multi - epitope antigen gene (mea) of Hantavirus’ s SEO type strain L99G2. Methods The amino acid sequence of G2 protein of L99 strain was analyzed and predicted by bioinformatics software, B cell epitopes were optimized, tandem epitopes of GPG sequences were introduced, mea was designed, then mea was constructed by overlapping PCR and cloned into prokaryotic The plasmid pET32a (+) was expressed. Results Five B cell epitopes were optimized. Mea was designed and successfully constructed. The recombinant plasmid pET32a-mea was obtained by PCR. Conclusion The recombinant hantavirus G2 glycoprotein mea and its prokaryotic expression system E.coli BL21 / pET32a-mea were constructed for the first time, which laid the foundation for its expression and immunological application.