Development and characterization of a recombinant cDNA-based hepatitis C virus system.
Invention of subgenomic HCV replicon a few years ago and recent success of in vitro production of infectious HCV have improved our knowledge of the HCV life cycle, replication, pathogenesis, and screening of anti-HCV therapeutics. However, the highly genotype-dependent nature of the in vitro HCV production system has limited its potential for HCV research. In this study, we constructed a recombinant DNA-based HCV system that contained EF-1alpha promoter-driven HCV genotype 1b with HCV E1/E2 deleted and replaced by GFP. We co-transfected this recombinant cDNA with HCV E1/E2 or VSV-G expression plasmid into 293T cells, and we showed HCV protein expression and processing and demonstrated production of HCV-like particles in culture supernatant of co-transfected cells. These results support potential use of this system for studies on expression and processing of the HCV polyprotein and assembly and release of HCV-like particles.
Ndjomou
Jean
J
Department of Microbiology and Immunology, Indiana University School of Medicine, R2 302, 950 W. Walnut Street, Indianapolis, IN 46202, USA.
Liu
Ying
Y
O'Malley
John
J
Ericsson
Maria
M
He
Johnny J
JJ
D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D016174:Hepacivirus / Q000502:physiology*; D006801:Humans; D007668:Kidney / Q000378:metabolism* / Q000821:virology*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014771:Virion / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D011994:Recombinant Proteins