Replication and Expression of HCV la／lb Chim eric Genome in Transfected HepG2 Cells

Abstract：T0 investigate the replication and expression of Hepatitis C virus(HCV)1 a and lb chimeric cDNA in HepG2 cells．HCV l a／ l b chimeric CDNA was used to construct an expression plasmid for the tran sfection of HepG2 cells．HCV protein and HCV genomic RNA an d an ti—genomic RNA in the tran sfected HepG2 cells an d the culture supernatan ts were detected by immunocytochemical staining， W estern blotting an d RT．PCR respectively．T|le results showed that HCV NS3 protein with about 70kI)a was detectable in the tran sfected HepG2 cells．HCV genomic RNA and an ti—genomic RNA were found positive both in the HepG2 cells an d the culture supernatan ts for more than 20 generations．HCV la／lb chimera Can replicate an d express in HepG2 cells．suggesting that the RNA—dependent RNA polymerase (RdRp)of HCV lb Can initiate the replication of HCV containing genotype la untran slated region(UTR)． The alterations of 5 UTR of HCV lb at nucleotides ll，12，l 3，34 and 35 dO not influence its binding to ribo some．The 3 UTR at nucleotides 9400，9403 and 9407，the deletion ‘ at nucleotide 9439 an d insertions ‘1fr’an d ‘AAT’at nucleotides 9409，9410 and 9495，9496，9497，do not significan tly influence the RdRp binding an d activities of HCV lb．ThiS HCV chimeric cDNA Can be Of value in the studies of HCV replication an d expression．