【摘 要】
：
Aims: Recently, robust Hepatitis C virus (HCV) genotype 2a (JFH1) and intrageontypic 2a/2a genome (J6JFH1) cell culture systems were developed.Since Nonstru
【机 构】
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Department of Microbiology, PLA Key Laboratory of Medical Microbiology, Second Military Medical Univ
【出 处】
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2008病毒性肝炎成就与挑战国际研讨会(International Confernece on Viral Hepati
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Aims: Recently, robust Hepatitis C virus (HCV) genotype 2a (JFH1) and intrageontypic 2a/2a genome (J6JFH1) cell culture systems were developed.Since Nonstructural 5A protein is supposed to be a pleiotropic protein with key roles in both viral RNA replication and infection, we aimed at exploiting the unique function of NS5A in the robust replication of J6JFH1.Methods: Intergeontypic chimera FL-J6JFH/J4 NSSA harboring HCV 1b NS5A domain were constructed by using overlapping PCR method.Huh7.5.1 cells were transfected with RNA transcripts of chimeric and wild type FL-J6JFH, negative control FL-J6JFH (GND).Cells were passaged and cell supernatants were harvested for infection assay.Cultures were examined for HCV protein expression(Immunofluorescence assay IFA, Western blot), HCV RNA titers (real-time PCR), and infectivity titers (50% tissue culture infectious dose).
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