截短的乙型肝炎表面抗原的原核表达及其特性的研究  

Prokaryocytic expression of the shortened hepatitis B surface antigen and antigenic analysis

作　　者：凌宁[1] 任红[1] 彭明利[1] 许红梅[1] 卿玉玲[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所,重庆400010

出　　处：《中华肝脏病杂志》2003年第4期209-211,共3页Chinese Journal of Hepatology

摘　　要：目的 在原核系统中表达截短的乙型肝炎表面抗原(HBsAg),并检测其抗原特性。为乙型肝炎疫苗(主要是治疗性疫苗)免疫原的选择进行初步的研究和探讨。 方法 利用聚合酶链反应(PCR)特异性扩增编码HBsAg羧基末端152和124个氨基酸的基因片段,均定向克隆至质粒pET32a(+),并将重组质粒分别转染至大肠杆菌(BL21)。 结果 重组质粒均成功构建。大肠杆菌表达产物氨基末端有1个含6个组氨酸肽段的150个氨基酸融合部分,以包含体形式存在。用SDS-PAGE、Western blot、ELISA等方法对原核表达产物进行检测,结果表明原核表达产物具有HBsAg反应原性。 结论 截短的HBsAg能在原核系统中表达,可为后续研究做准备。Objective To study the expression of the shortened hepatitis B surface antigen in prokaryocyte and detect the antigenic characters. Methods Firstly, the gene fragments coding the 152 and 124 amino acids of the carboxyl terminus of hepatitis B surface antigen (HBsAg) were amplified by polymerase chain reaction (PCR). Secondly, they were cloned to plasmid pBKS+, and the accuracy of those constructions were confirmed by restriction enzyme digestion and DNA sequencing. Then, they were cloned to prokaryocytic expression vector-plasmid pET32a(+). The recombinant plasmids were transfected into E.coli BL21 and induced to express with IPTG. Results The recombinant plasmids were successfully constructed. In E.coli BL21, the protein was expressed in a fusion fashion and could be recognized by monoclonal antibody against HBsAg with ELISA and Western blot. Conclusion The shortened HBsAg can be expressed in prokaryocyte.

关 键 词：乙型肝炎表面抗原 乙型肝炎疫苗 基因 原核细胞 

分 类 号：R512.620.3[医药卫生—内科学]