Analysis of virus adaptations in the novel HCV cell culture and infection system

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We have established HCV culture system using JFH-1 strain isolated from a fulminant hepatitis patient. This system enabled us to analyze HCV life cycles in detail. In this research project, we have analyzed how HCV adapted in cultured cells and organisms. We have also tried to determine which virus components involved in HCV adaptation.HCV adaptation in cultured cells: we transfected in vitro-synthesized HCV RNA into Huh7 cells and collected infectious HCV particles. HCV infection sometimes causes plaque formation. We collected plaque forming HCVs, and infected to naive Huh7 cells. Isolated virus subclones from individual plaque showed greater replication efficiency and cytopathogenicity than parental virus. We analyzed the sequence of this HCV and found several nucleotide mutations with amino acid substitution in NS5B coding region. These mutations may be involved in plaque forming ability and virus genome replication.