Aim: To express HCV F protein gene fragment in E.coli and detect if there is its antibody in HCV patients.
Methods: RNA of the virus identified as genetype 1b was choosed as template, the F protein gene was amplified by RT-PCR. This gene fragment was inserted into plasmid vector pGEM simple T. F gene was digested by EcoR I and BamH I from pGEM and cloned into plasmid vector 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 the patients which HCV RNA was positive by ELISA.
Results: After IPTG induction, a positive band about 43 kD was detected. 82 of 120 HCV RNA positive's sera had anti-F antibody by ELISA.
Conclusion: It is possible to efficiently express the HCV F protein in E.coli. The positive rate of anti-F antibody in HCV RNA positive patients was 68%.