Construction and Expression of Eukaryotic Expression Plasmid of Core Gene of Hepatitis C Virus

Objective:To construct the eukaryotic expression plasmid of core gene of hepatitis C virus(HCV)and to express it in HepG2 cells.Methods:The cDNA fragment of HCV core region was synthesized by the reverse-transcriptase-polymerase chain reaction (RT-PCR)from the sera of patients with hepatitis C and cloned into the eukaryotic expression vector pcDNA3.0 to construct the plasmid pcDNA-HCV,which was identified by cutting with restriction endonuclease and sequencing.The plasmid pcDNA-HCV was then trans- fected into HepG2 cells and screened by G-418.The expression of HCV core protein in HepG2 cells was analyzed by Western Blotting and immunocytochemical technique.Results:503 bp cDNA fragment of HCV core gene,which was obtained from the sera of patients with hepatitis C,belonged to C region of genotype 1 of HCV.By Western Blotting analysis and immunocytochemical technique,it was found that there was the expression of HCV core protein in HepG2 cells.Conclusions:Recombinant eukaryotic expression plasmid pcD- NA-HCV is successfully constructed and stably expresses in HepG2 cells.