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目的:在大肠杆菌中对登革2型病毒包膜蛋白B区进行表达,为观察其抗原性和免疫原性奠定基础。方法:将通过PCR扩增的登革2型病毒包膜蛋白B区插入高效原核表达载体pBAD/TopoThioFusion,然后在大肠杆菌中利用阿拉伯糖进行诱导表达。对表达产物以免疫印迹和ELISA法进行检测。结果和结论:登革2型病毒包膜蛋白B区在大肠杆菌中的表达产物以可溶性形式存在,表达量约占细菌可溶性总蛋白的62%。表达产物可与登革2型病毒的特异多抗反应,但与登革1、3和4型病毒的特异多抗无交叉反应。登革2型病毒包膜蛋白B区可在大肠杆菌中以可溶性形式高效表达。
OBJECTIVE: To express dengue type 2 envelope protein B region in Escherichia coli and lay the foundation for its antigenicity and immunogenicity. Methods: The dengue type 2 envelope protein B region amplified by PCR was inserted into the highly efficient prokaryotic expression vector pBAD / TopoThioFusion and then induced by arabinose in E. coli. The expression products were detected by Western blotting and ELISA. RESULTS AND CONCLUSION: The expression product of dengue 2 envelope protein B region in E. coli was in soluble form and accounted for about 62% of the total bacterial soluble total protein. The products of expression could react with specific polyclonal antibody against dengue 2 virus but no cross-reaction with specific polyclonal antibodies to dengue 1,3 and 4 viruses. Dengue virus type 2 envelope protein B region can be efficiently expressed in soluble form in E. coli.