乙型肝炎病毒重组e抗原在大肠杆菌中的高效表达及初步应用

HIGH EXPRESSION OF RECOMBINANT HEPATITIS B VIRUS e ANTIGEN IN E. COLI AND ITS PRELIMINARY APPLICATION

为了获得重组表达乙型肝炎病毒 ｅ抗原（ＨＢｅＡｇ）。利用聚合酶链反应（ＰＣＲ）扩增和重组技术，在大肠杆菌中的高表达非融合型重组ＨＢｅＡｇ。结果：为模拟天然ＨＢｅＡｇ的结构，除第一个氨基酸为起始密码子ＡＴＧ所编码的甲硫氨酸外，共包括Ｐｒｃｅ－Ｃ区羧基端的９个氨基酸和ＨＢｃＡｇ Ｎ－末端的１４９个氨基酸残基、经诱导表达，其产物相对分子质量约为１７．５ × １０３，产量占细菌总蛋白的１１％。结论：用Ｗｅｓｔｅｒｎ印迹法分析，本表达产物可与特异性的ＨＢｅＡｇ抗体反应。用斑点酶免疫法可以检测患者血清的抗ＨＢｅ。

PURPOSE To obtain a large amount of recombinant hepatitis B virus e antigen (rHBeAg) for the replacement of serium HBeAg purificed from inpatient sera. METHODS By means of polymerase chain reaction (PCR) and recombinant technics, the rHBeAg was expressed as nonfusion protein in E. Coli. RESULTS To mimic the natural structure of HBeAg, except the first amino acid (methionine) was added by ATG, the construct included 9 amino acid residues at the C-terminus of the Pred region and 149 amino acid residues at the N-terminus of the core gene. After induction, the molecular weight of the recombinant protein was around 17.5 × 103, and the amount of recombinant protein was estimated as l 1 % of the whole bacterial lysate protein. CONCLUSIONS This rHBeAg product expressed in E. Coli could specifically react with human anti-HBe serum by analysis of western blotting and could be used to detect anti-HBe in patient sera by dot enzyme immunoassay.