ABSTRACT
Abnormal cell signal transduction is associated with the occurrence and development of human diseases. Some virus pathogenicity and infection mechanism are due to virus antigen protein acting on the host cell signal transduction pathway, leading to host cell signal transduction disorder. Hepatitis C virus (HCV) is a major pathogen of chronic hepatitis C, which causes cirrhosis and hepatocellular carcinoma. But the pathogenesis of HCV and persistent infection mechanism remain far from clear. HCV pathogenesis may be related to the HCV protein expression interfering host cell signal transduction pathways. The studies of hepatitis C virus proteins acting on host cell signal transduction pathways, not only help to clarify the impact of its pathogenic mechanisms, but also benefit to new drug design and development for new treatment methods. This article summarizes the recent progress in research on the effect of hepatitis C virus protein in cell signal transduction pathways in the past few years.
ABSTRACT
Objective To construct the eukaryotic expression vector coding HCV gene E2 fused with His-Tag, and to express fused protein in CHO cells for investigating the function of HCV envelope protein E2. Methods The gene encoding HCV envelope protein E2 was amplified from pBRTM/HCV1-3011, a plasmid containing the cDNA of HCV's ORF, by polymerase chain reaction (PCR) method and cloned into the vector pET28(a) containing His-Tag to obtain the fused HCV envelope protein E2 gene fused with His-Tag. The fused gene was cloned into pcDNA3.1 to construct the recombinant plasmid pcDNA3.1-His-E2, which will express the E2 protein, fused with His tag. This recombinant plasmid was transfected into CHO cells by Lipofactamine 2000 reagent. The fused protein was identified by indirect immunofluorescence (IIF) and Western-blot (WB) methods. Result The positive results were obtained when the fused protein of HCV E2 with His-Tag were identified by IIF and WB methods. Conclusion The eukaryotic expression vector pcDNA3.1-His-E2 was constructed successfully and the fused proteins were expressed in cells.
ABSTRACT
To screen and characterize human phage antibody to hepatitis C virus E2 antigen. The recombinant phages were panned by HCV E2 antigen which was coated in a microwell plate. After three rounds of biopanning, 56 clones specific to HCV E2 antigen were obtained. The specificity of scFv was determined by ELISA method and cross reaction with BSA and competitive inhibition assay. HCV E2 phage antibody had a specific combination character with recombinant hepatitis C virus E2 antigen. The DNA sequence data showed that the scFv coding gene was 771bp in size