title: Subgenomic replicon derived from a cell line infected with the hepatitis C virus.
abstract: Recently, cell culture systems have been established, where a hepatitis C virus (HCV) subgenomic replicon was efficiently replicated and maintained for a long period. To see whether a HCV sequence derived from HCV-infected cultured cell sequence can be used for the construction of a functional replicon, a HCV subgenomic RNA carrying a neomycin-resistant gene was constructed using the HCV genome RNA obtained from cultured cells infected with HCV. After transfection, G418-resistant Huh-7 cells were selected and subcloned. Finally, the production of HCV proteins and de novo synthesis of subgenomic RNA were confirmed in the selected cell clone, indicating that this subgenomic RNA replicated in cultured cells and functioned as a replicon. These results suggest that the HCV genome obtained from an in vitro HCV infection system with cultured cells can be used to develop a subgenomic replicon system with diverse HCV sequences.
authors.affiliation: Department of Viral Oncology, Institute for Virus Research, Kyoto University, 53 Kawara-cho Shogo-in, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Hiroe
authors.initials: H
authors.lastname: Kishine
authors.forename: Kazuo
authors.initials: K
authors.lastname: Sugiyama
authors.forename: Makoto
authors.initials: M
authors.lastname: Hijikata
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Kato
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Takahashi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Noshi
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Nio
authors.forename: Masahiro
authors.initials: M
authors.lastname: Hosaka
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Miyanari
authors.forename: Kunitada
authors.initials: K
authors.lastname: Shimotohno
mesh_terms: D002460:Cell Line; D002999:Clone Cells; D016679:Genome, Viral; D005839:Gentamicins / Q000494:pharmacology; D016174:Hepacivirus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D006801:Humans; D009154:Mutation; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics; D012093:Replicon; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D014779:Virus Replication
chemical_list: D005839:Gentamicins; D012367:RNA, Viral; D014764:Viral Proteins; C010680:antibiotic G 418