PMID: 9186943May 1, 1997Paper

Construction of hepatitis C-SIN virus recombinants with replicative dependency on hepatitis C virus serine protease activity

Journal of Virological Methods
Y G ChoY C Sung

Abstract

An in vivo assay system was developed for the serine protease of hepatitis C virus (HCV) using the sindbis (SIN) viral replication system in which HCV serine protease activity is essential for the replication of the HCV-SIN chimeric virus. Two chimeric viral cDNA clones were constructed by inserting the NS3/4A region and NS3/4A region with the putative helicase deleted, into the N-terminal region of SIN core protein. The constructs were named Tpro CT and Tpro T, respectively. BHK-21 cells transfected with the in vitro transcribed RNAs from Tpro CT and Tpro T showed specific cytopathic morphology and produced chimeric viruses, Vpro CT and Vpro T. In contrast, in vitro transcribed RNAs from Tpro CTI and Tpro TI, in which serine of catalytic triad of HCV protease was changed to alanine, were not infectious. When the chimeric viruses were passaged in BHK-21 cells at about 0.1 multiplicity of infection (MOI), Vpro T, but not Vpro CT, stably expressed HCV protease for up to five passages. Surprisingly, the cell culture media of BHK-21 cells infected with Vpro T, compared to wild-type sindbis virus, showed rapid pH changes by more than 0.8 pH degree at 72 h post-infection. HCV-SIN hybrid viruses could be used in screening the HCV prot...Continue Reading

References

Jan 1, 1992·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·H HagiwaraT Kamada
Apr 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·C S HahnC M Rice
Mar 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·R H Miller, R H Purcell
Sep 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·I SaitoY Ohta
Feb 1, 1994·Proceedings of the National Academy of Sciences of the United States of America·E PizziR De Francesco

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Citations

Jul 22, 1998·Journal of Virological Methods·Y G ChoY C Sung
Dec 24, 1998·Antiviral Research·A D KwongS A Raybuck
Mar 29, 2005·Antimicrobial Agents and Chemotherapy·Victoria ChungEric A D'Souza
Jul 30, 2008·Expert Review of Anti-infective Therapy·Arthur Y Kim, Joerg Timm
Nov 8, 2003·World Journal of Gastroenterology : WJG·Hong-Xia MaoZheng-Hong Yuan
Oct 10, 1998·Clinical Microbiology Reviews·A K Patick, K E Potts

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