丙型肝炎病毒河北定州株C33c基因的克隆、序列分析及在大肠杆菌中的表达

Cloning, sequencing and expression in Escherichia.coli of HCV (Hebei Dingzhou strain) C33c gene

采用反转录ＰＣＲ方法从河北定州地区献血员血清中克隆了丙型肝炎病毒（ｈｅｐａｔｉｔｉｓＣｖｉｒｕｓ，ＨＣＶ）的Ｃ３３ｃ抗原基因，序列分析结果表明，该株的Ｃ３３ｃ基因和中国泰安株的同源性为９１．０％，氨基酸序列同源性为９２．２％；与ＨＣＶ河北株的同源性分别为９１．３％和９６．２％；和日本ＪＫ１株的同源性分别为９１．６％和９４．８％。克隆的基因在大肠杆菌中得到表达和纯化。

Gene encoding hepatitis C virus C33c antigen was cloned by reversetranscription followed by polymerase chain reaction (RTPCR) using RNA template isolated from antiHCV positive serum of a blood donor from Dingzhou, Hebei Province. DNA sequencing showed 91% homology with Chinese Taian strain and 91.6% with Japanese JK1 strain, respectively. The deduced amino acid homology with these two strains was 92.2% and 94.8% respectively. The C33c gene was expressed in Escherichia coli, and the purified C33c antigen showed to be immunodominant since it could react with C33c positive sera, and was successfully used as the component in secondgeneration antiHCV ELISA kits.