丙型肝炎病毒ns2基因的克隆及其表达.pdfVIP

第 21 卷 3 期 中 国 病 毒 学 21 （3 ）：217-220 2006 年 5 月 VIROLOGICA SINICA May 2006 丙型肝炎病毒ns2 基因的克隆及其表达* 1 2 1** 刘超红 ，焦成松 ，陈新文 (1.中国科学院武汉病毒研究所 病毒学国家重点实验室，湖北武汉 430071 ；2.广州军区武汉陆军总医院，湖北武汉 430071) Cloning of HCV ns2 Gene and its Expression in Prokaryotic and Mammalian cells 1 2 1** LIU Chao-hong ，JIAO Chen-song ，CHEN Xin-wen (1.State Key Laboratory of Virology, Institute of Virology, Chinese Academy of Sciences， Wuhan 430071, China ；2. Wuhan General Hospital of Guangzhou Command. Wuhan 430071,China ) Abstract ：The full-length and truncated ns2 gene (2881-3078 bp) were gained through PCR using p90-HCV as template. The truncated ns2 gene was cloned into the prokaryotic expression vector PET32a. The fusion protein Trx-His-NS2C expressed in a soluble form in E.coli, and then was purified by His resin. Highly specific and efficient antibody was produced by immunizing the rabbit with purified fusion protein. In addition to this, recombinant adenovirus containing the full-length ns2 gene was constructed. The NS2 protein was expressed successfully in a size of 23.2kDa by infecting 293 and HepG2 cells with recombinant adenovirus. These results had set a base for the further stulies on the structure and function of the NS2 protein. Key words ：NS2A ； Hepatitis C virus （HCV ）；Antibody; Expression ；Adenovirus 摘要：以p90-HCV 为模板，通过 PCR 分别得到全长和截短（2881-3078 bp ）ns2 基因。将截短 ns2 基因克隆到 pET32a