乙型肝炎病毒adr亚型X蛋白在大肠杆菌中的高效表达

High Level Expression of Hepatitis B Virus adr Subtype X Protein in Escherichia coli

将ａｄｒ亚型ＨＢＶ的完整Ｘ基因克隆到表达质粒ｐＰｒｏＥＸＨＴｂ中，实现了Ｘ蛋白在大肠杆菌中的高效融合表达。表达产物全长１７９个氨基酸残基（其中Ｘ蛋白部分１５４个氨基酸残基），分子量约１９．７ｋＤ，约占细菌总蛋白的３４％。其氨基端融合部分含有６组氨酸肽段，经ＨｉｓＴｒａｐＴＭ亲和层析纯化，一步可达纯度９３％。Ｗｅｓｔｅｒｎ印迹分析表明，该表达产物可与ＨＢＶ感染患者体内的抗Ｘ抗体特异性结合。

The X gene of a known adr subtype of hepatitis B virus (HBV) was cloned into the ProEX HT Prokaryotic Expression System(GibcoBRL),and the X protein of HBV,154 amino acid residues,was expressed in a fusing fashion in E.coli . The fusion protein,named His tagged X protein (shortly HX protein) contains 179 amino acid residues and its molecular weight is about 19.7 kilodaltons.It is easy to purify by the HisTrap affinity chromatography. The purity of the HX protein could reach 93%by one step of capturation.Western blotting showed that the HX protein could specifically be recognized by the human antibodies against HBV X antigen(ptotein).