title: Synthesis of encephalomyocarditis virus in a cell-free system: from DNA to RNA virus in one tube.
abstract: Virus particles are used in vaccination, drug delivery, and material sciences. Here we devised a system where the RNA virus encephalomyocarditis virus (EMCV) is synthesized from DNA templates in vitro. When a plasmid or a PCR product harboring the full-length cDNA of EMCV in the T7 promoter/terminator unit was incubated in a HeLa cell extract supplemented with T7 RNA polymerase, EMCV was produced within 4 h at an efficiency of over 10-fold compared with the system programmed with the EMCV RNA. The EMCV RNA transcribed by the virally encoded RNA-dependent RNA polymerase was predominantly incorporated into the EMCV particle even in the presence of a larger amount of the EMCV RNA transcribed by T7 RNA polymerase from the plasmid.
authors.affiliation: Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo, Himeji, 671-2280, Japan.
authors.forename: Tominari
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Satoshi
authors.initials: S
authors.lastname: Mikami
authors.forename: Mamiko
authors.initials: M
authors.lastname: Masutani
authors.forename: Kodai
authors.initials: K
authors.lastname: Machida
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Imataka
mesh_terms: D001709:Biotechnology / Q000379:methods; D002457:Cell Extracts; D002474:Cell-Free System; D004686:Encephalomyocarditis virus / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids; D012367:RNA, Viral / Q000096:biosynthesis*; D014764:Viral Proteins / Q000096:biosynthesis*; D014773:Virology / Q000379:methods; D019065:Virus Assembly
chemical_list: D002457:Cell Extracts; D012367:RNA, Viral; D014764:Viral Proteins