Analysis of persistent infection mechanism of RNA virus

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One of the major impediments to the structural analysis of the HCV genome and genetic analysis of viral replication has been lack of a reliable cell culture system permissive for HCV replication. Therefore, we established the HCV replication system by using entire HCV cDNA and T7 RNA polymerase recombinant adeno virus system.Methods : the putative full-length HCV genome (1-9603) was transiently expressed by the adeno virus/T7 RNA polymerase hybrid expression system. We transfected to a HCV high sensitive cell line (IMY cell) with HCV entire cDNA clone and infected recombinant adeno virus. The HCV particle was characterized by immuno-electron microscopy. Results : The transfected HCV genome replicated in cells, as evidenced by appearance of progeny HCV RNA and detection of negative-strand viral RNA.Immuno-electron microscopic studies have shown that the HCV like-particle was detected in the transfected cells and culture medium. We show the replication process of HCV in the cell and clarified the structure of viral particle. Conclusion : The expression system using HCV entire cDNA clone and showed the importance of 3'X region for HCV replication in the cell. The elctron microscopy study revealed the dynamics of viral core particle in the cells. Moreover the core particle was proved to fold the hexagonal structure.