title: A novel reverse genetics system for production of infectious West Nile virus using homologous recombination in mammalian cells.
abstract: Reverse genetics systems facilitate investigation of many aspects of the life cycle and pathogenesis of viruses. However, genetic instability in Escherichia coli has hampered development of a reverse genetics system for West Nile virus (WNV). In this study, we developed a novel reverse genetics system for WNV based on homologous recombination in mammalian cells. Introduction of the DNA fragment coding for the WNV structural protein together with a DNA-based replicon resulted in the release of infectious WNV. The growth rate and plaque size of the recombinant virus were almost identical to those of the parent WNV. Furthermore, chimeric WNV was produced by introducing the DNA fragment coding for the structural protein and replicon plasmid derived from various strains. Here, we report development of a novel system that will facilitate research into WNV infection.
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, N18, W9, Kita-ku, Sapporo 060-0818, Japan. Electronic address: shin-kobayashi@vetmed.hokudai.ac.jp.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, N18, W9, Kita-ku, Sapporo 060-0818, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Yoshii
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, N18, W9, Kita-ku, Sapporo 060-0818, Japan.
authors.forename: Minato
authors.initials: M
authors.lastname: Hirano
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, N18, W9, Kita-ku, Sapporo 060-0818, Japan.
authors.forename: Memi
authors.initials: M
authors.lastname: Muto
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, N18, W9, Kita-ku, Sapporo 060-0818, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kariwa
mesh_terms: D000818:Animals; D002460:Cell Line; D057809:HEK293 Cells; D059765:Homologous Recombination; D006801:Humans; D012093:Replicon; D059386:Reverse Genetics / Q000379:methods*; D014779:Virus Replication / Q000235:genetics; D014901:West Nile Fever / Q000821:virology; D014902:West Nile virus / Q000235:genetics* / Q000254:growth & development / Q000502:physiology*
keywords: Homologous recombination; Reverse genetics; West Nile virus