EXPRESSION AND PRELIMINARY APPLICATION OF HCV F PROTEIN GENE

摘要：

To express HCV F protein gene fragment in E. coli and to confirm whether there was its antibody in the sera of patients with HCV infection. RNA of HCV identified as genetype 1b was choosed as template, the F protein gene was amplified by RT-PCR. This gene fragment was inserted into plasmid pGEM simple T. F gene was digested by EcoRI,BamHI from pGEM and cloned into plasmid pGEX-4T-2. The ligation mixture was transformed into E. coli. TG1 and F fragment expression was induced by IPTG. The expressed protein was purified from lysates with Glutathione Sepharose 4B. The purified protein was used to identify whether there was anti-F antibody in patients with HCV RNA positive by ELISA. After IPTG induction, a positive band about 43KD was detected and 82 of 120 (68. 3％) HCV RNA positives sera had anti-F antibody by ELISA, This study suggested that it was possible to efficiently express the HCV F protein in E. coli.

关键词： Hepatitis C vims F protein gene cloning protein expression polyclonal antibody

作者: Yun Zhang Chunmei Jiang Zhengyu Diao Xiaozhao Deng Ke Xu Weiliang Ding Yongfei Tan

作者单位: Institute of Military Medicine Nanjing Cammand PL At Nanjing 210002, China Department of Epidemiology and Biostatistics in Nanjing Medical University, Nanjing 210029, China Yixing People Hospital, Yixing 214200, China

会议类型: 国际会议

会议名称: The 4th International Forum on Post-genome Technologies(4IFPT)(第四届国际后基因组生命科学技术学术论坛)

会议地点: 杭州

会议语种:英文

页码: 492-494

在线出版日期: 2006-09-25（万方平台首次上网日期，不代表论文的发表时间）