乙型肝炎病毒外膜蛋白和核壳蛋白双表达质粒的构建和表达

Construction and expression of hepatitis B virus envelope protein combined with core protein with two multiple cloning sites vector

目的构建同时表达乙型肝炎病毒外膜蛋白和核心蛋白的真核表达质粒并研究其表达效果。方法利用DNA重组技术，构建由两个CMV启动子分别表达乙型肝炎病毒外膜蛋白和核心蛋白的双表达真核质粒vR—sc，体外真核表达后ELISA检测HBsAg和HBeAg。结果在乙肝病毒外膜蛋白表达质粒上插入多克隆位点，并插入乙肝病毒核心蛋白的完整表达调控单元，体外真核表达后成功检测到HBsAg和HBeAg。结论成功构建双表达质粒VR—SC，其表达效果良好，为进一步改造成HBVDNA疫苗打下了基础。

Objective To develop a coexpression plasmid which expressing envelope protein and nucleoprotein of hepatitis B virus and know of its expressing efficiency. Methods The plasmid coexpressing envelope protein and nueleoprotein of hepatitis B virus under the CMV promoter respectively was constructed by gene recombination. Cellular expression was assessed by ELISA. Results Multiple cloning site was inserted into expression vector contain hepatitis B virus PreS2-S gene. And expression unit containing hepatitis B virus PreC-C was cloned into it. HBsAg and HBeAg was detected both in the culture supematant and in the cells. Conclusion The coexpressing plasmid was constructed successfully and it can express effectively in vitro. This has provided a basis for further research of the therapeutic HBV DNA vaccine.