摘要
本文从两例临床确诊为乙型肝炎患者的2mg肝穿刺标本中利用基因扩增方法,设计扩增出缺失羧基端34个氨基酸的乙型肝炎病毒C抗原(HBcAg)基因的0.45kb基因片段,将此片段克隆入表达载体,导入大肠杆菌中表达,结果获得每毫升培养液含有100万酶联反应单位的HBgAg,也存在1万酶联反应单位的HBcAg。SDS-聚丙烯酰胺凝胶电泳显示,其表达产物占菌体总蛋白的30～37％,分子量为155kD。HBeAg基因结构及表达形式有待进一步研究和探讨。
The 0.45kb fragments, which were deleted 34 amino acid sequence of N terminus of human hepatitis B virus C gene, were obtained from two liver puncture samples of patient using polymerase chain reaction.The fragments were cloned to expression vector pBV220 and expressed in E.coli.The results showed there were a a 1 000 000 ELISA analystic unit of HBeAg and<10 000 of HB g in 1 mL culture of E.coli.The products of HBeAg had a 15.5 kD on SDS-PAGE.The expression amount of HBeAg reached 30~37% of total bacterial protein.The mechanism of HBeAg expression in vivo was discussed.
