Cloning and expression of core gene cDNA of Chinese hepatitis C virus in cosmid pTM3

AIM To clone core gene cDNA of Chinese hepatitis C virus (HCV) into eukaryotic expression vector cosmid pTM3 and to express HCV core antigen in HepG2 cells. METHODS Core gene cDNA of HCV was introduced into eukaryotic expression vector cosmid pTM3. Using vaccinia virus/bacterio phage T7 hybrid expression system, HepG2 cells were transfected with the recombinant plasmid pTM3-Q5 34 by lipofectin. RESULTS From the transfected bacteria Top10F’, 2 pTM3-Q534 c lones containing the recombinant plasmid were identified from randomly selected 10 ampicillin-re sistant colonies. By reverse transcription PCR and indirect imm unofluorescence technique, HCV RNA and core protein was identified in HepG2 cells transfected with the recombinant plasmid. CONCLUSION The construction of a recombinant plasmid and the expression of core gene cDNA of HCV in HepG2 was successful.