中华医学杂志（英文版）2002，Vol.115Issue(5)：722-725,4.
乙型肝炎病毒e抗原前体蛋白在大肠杆菌中的克隆、表达和纯化及其特性的初步研究
Cloning and expression and purification of Hepatitis B e-antigen precursor in Escherichia coli
摘要
Abstract
Objective To investigate the role of the 25 kD hepatitis B e antigen (HBeAg) precursor tha t only exist inside hepatocytes and study its effect on the pathopoiesis of hep jatitis B and QIAGEN expression and purification system.Methods Hepatitis B virus(HBV) preC/C gene for the 25 kD HBeAg precursor was cloned into the expression vector pQE30 and the 25 kD HBeAg precursor was expressed in Esch erichia coli (E.coli) and purified. Its antigenicity for 21 kD mature HBeAg wa s tested by western blot analysis. Results Cloned fragments in the expression vector were sequenced and verified to be homo genous with that of HBV (ayw subtype ). Expression of the HBeAg precursor in E .coli under the transcriptional regulation of T5 promoter yielded a soluble cyt osolic protein with an apparent molecular mass of 25 kD. Recombinant HBeAg prec ursor exhibited identical potencies with 21 kD mature HBeAg that reacted with an ti-HBeAg antibodies. The purification rate of the expressed HBeAg precursor wa s up to 89.6% and the yield of purified HBeAg precursor from this procedure was 2.4 mg/L. Conclusion 25 kD HBeAg precursor exhibited biological activity and might play an important role in pathopoiesis of hepjatitis B.关键词
HBeAg前体蛋白 乙型肝炎病毒 pQE30Key words
HBeAg precursor/Hepatitis Bvirus/pQE30分类
医药卫生引用本文复制引用
周福元,隋礼丽,骆抗先,侯金林..乙型肝炎病毒e抗原前体蛋白在大肠杆菌中的克隆、表达和纯化及其特性的初步研究[J].中华医学杂志（英文版）,2002,115(5):722-725,4.基金项目
The work was supported by grants from the National Natural Science Foundation o f China (No.39630280). （No.39630280）
