登革2型病毒包膜蛋白B区在大肠杆菌中的高效表达

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栽体pBAD／Topo ThioFusion，然后在大肠杆菌中利用阿拉伯糖进行诱导表达。时表达产物以免疲印迹和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。

Highly expression of B domain of dengue virus type 2 envelope protein in E.coli

Objective: To express B domain of dengue virus type 2 envelope protein in E.coli. Methods: The gene of B domain of dengue virus type 2 envelope protein, which was amplified by PCR, was inserted into prokaryotic expression vector pBAD/Topo ThioFusion. Then expression was induced with arabinose in E.coli. The expression products were detected by Western blotting and ELISA. Results and Conclusion: The expression products were soluble, and the yields were 62% of total soluble bacterial proteins. The expression products could specifically react with polyantibody against dengue virus type 2, but could not react with other three serotypes. B domain of dengue virus type 2 envelope protein could be highly expressed in E.coli.