Construction of Mammalian Expression Vector of Core Gene of HCV of Pakistani Isolate Genotype 1a

Abstract

Background: Cell lines are a valuable tool to identify the HCV virus infection and propagation and by establishing cell lines expressing core protein we can explore the role of HCV core protein in development of HCC. Results: The results indicate PCR amplification of 573bp product of core gene and sequencing confirmed the 1A genotype of HCV of Pakistani isolate. Then recombinant plasmid was digested with restriction enzymes (Hindi III and EcoR I) which gave fragments of 5.5 kb and 0.6 kb and It was a prove that our plasmid having a core gene. 21KDa core protein extracted from the transiently transfected Huh 7 cell line with recombinant mammalian vector of core gene was detected by using anti-core monoclonal antibody. Conclusion: we successfully constructed a mammalian expression vector of core gene of Pakistani isolate of genotype 1A that was encoding 21KDa of core protein in Huh 7 cell line.