HBV核心和前S1融合蛋白在大肠杆菌中的表达及真核表达载体的构建

目的:表达可用于乙型肝炎治疗性疫苗研究的乙型肝炎病毒(HBV)核心抗原(HBcAg)与前S1抗原(PreS1)氨基酸(AA)3～55肽段融合蛋白。方法:以原核表达载体pET-11d表达HBV融合蛋白,采用免疫印迹技术和酶联免疫吸附试验(ELISA)分析表达产物的抗原性。构建以CMV为启动子的表达HBV核心和前S1融合蛋白的真核表达载体。结果:HBV融合蛋白在大肠杆菌中有效稳定表达,表达产物具有核心抗原和前S1特异杭原性。成功构建了pCI-CS1真核表达载体。结论:本方法有效表达了HBV融合蛋白,表达产物具有特异抗原性。融合蛋白的表达及真核表达载体的构建为进一步进行HBV CS1融合蛋白和基因疫苗研究奠定了基础。

Expression of Fusion Protein of HBV Core and PreS1 Gene in E. coli.

Objective: To express the fusion protein of HBV core plus PreSl for development of HBV vaccine. Methods: pET-11d was used to express the fusion protein which was comprised of partial region (amino acids 3 - 55) of PreSl introduced at the C-terminal part of HBV core truncated protein (amino acids 1 - 155). The antigenicity of fusion protein was analysed by Western blot and EIA. The HBV core and PreSl gene was also cloned under the CMV promoter in eukaryotic expression vector. Results: Fusion protein of HBV was efficiently expressed in E. coli. The expressed product possessed special antigencity of both HBcAg and PreS1. Eukaryotic expression vector for HBV Core plus PreSl was constructed. Conclusion: This fusion protein which possesses special antigencity of both HBcAg PreSl may be developed into vaccine for the therapy of chronic hepatitis B. pCI-CS1 vector will be used for further study on the potential role of HBV CS1 as therapeutic vaccine.