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Advantages of a single-cycle production assay to study cell culture-adaptive mutations of hepatitis C virus.


ABSTRACT: The JFH1 strain of hepatitis C virus (HCV) is unique among HCV isolates, in that the wild-type virus can traverse the entire replication cycle in cultured cells. However, without adaptive mutations, only low levels of infectious virus are produced. In the present study, the effects of five mutations that were selected during serial passage in Huh-7.5 cells were studied. Recombinant genomes containing all five mutations produced 3-4 logs more infectious virions than did wild type. Neither a coding mutation in NS5A nor a silent mutation in E2 was adaptive, whereas coding mutations in E2, p7, and NS2 all increased virus production. A single-cycle replication assay in CD81-deficient cells was developed to study more precisely the effect of the adaptive mutations. The E2 mutation had minimal effect on the amount of infectious virus released but probably enhanced entry into cells. In contrast, both the p7 and NS2 mutations independently increased the amount of virus released.

SUBMITTER: Russell RS 

PROVIDER: S-EPMC2393785 | biostudies-literature | 2008 Mar

REPOSITORIES: biostudies-literature

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Advantages of a single-cycle production assay to study cell culture-adaptive mutations of hepatitis C virus.

Russell Rodney S RS   Meunier Jean-Christophe JC   Takikawa Shingo S   Faulk Kristina K   Engle Ronald E RE   Bukh Jens J   Purcell Robert H RH   Emerson Suzanne U SU  

Proceedings of the National Academy of Sciences of the United States of America 20080311 11


The JFH1 strain of hepatitis C virus (HCV) is unique among HCV isolates, in that the wild-type virus can traverse the entire replication cycle in cultured cells. However, without adaptive mutations, only low levels of infectious virus are produced. In the present study, the effects of five mutations that were selected during serial passage in Huh-7.5 cells were studied. Recombinant genomes containing all five mutations produced 3-4 logs more infectious virions than did wild type. Neither a codin  ...[more]

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