【摘要】 目的构建一种能够激发特异性细胞免疫功能的基因工程乙肝疫苗。方法选取HBV核心抗原18-27氨基酸CTL表位,用PCR的方法融合在结核杆菌热休克蛋白HSP65下游,并将此融合基因在大肠杆菌表达系统pET15b中表达。结果 已经构建出热休克蛋白-乙型肝炎核心抗原表位(HSP-HBV-ME)融合基因,并在大肠杆菌BL21中表达出融合蛋白,经SDS-PAGE和Western印迹分析证明所表达的蛋白质是HSP-HBV-ME融合蛋白。结论利用PCR法构建融合基因是一种简单、快速的方法,而且可以在构建过程中对密码子进行优化,从而达到高效表达重组蛋白的目的。更多还原

【Abstract】 Objective To construct the fusion gene of a genetically-engineered HBV vaccine which can stimulate the specific cellular immune function against HBV. Methods HBcAg CTL epitope 18-27 was fused into the downstream of BCG heat shock protein 65 by using PCR and the fusion protein was expressed in E. coli expression system pET15b. Results Fusion gene of HSP-HBV-ME was constructed and expressed in E. coli strain BL21 and it was confirmed by SDS-PAGE and Western blot that the protein expressed is HSP-HBV-ME. Conclusion It is easy and rapid to construct fusion genes by using the PCR and the genetic code used can be optimized to enhance the expression of recombinant protein.更多还原