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/bacteriophage T7 hybrid expression system, HepG2 cells were transfected with the recombinant plasmid pTM3-Q534 by lipofectin.RESULTS:From the transfected bacteria Top10F', 2 pTM3-Q534 clones containing the recombinant plasmid were identified from randomly selected 10 ampicillin resistant colonies. By reverse transcription PCR and indirect immunofluorescence 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.
Jiang
Rong-Long
RL
Lu
Qiao-Sheng
QS
Luo
Kang-Xian
KX