title: Purification and visualization of encephalomyocarditisvirus synthesized by an in vitro protein expression system derived from mammalian cell extract.
abstract: Virus particles are promising vehicles and templates for vaccination, drug delivery and material sciences. Although infectious picornaviruses can be synthesized from genomic or synthetic RNA by cell-free protein expression systems derived from mammalian cell extract, there has been no direct evidence that authentic viral particles are indeed synthesized in the absence of living cells. We purified encephalomyocarditis virus (EMCV) synthesized by a HeLa cell extract-derived, cell-free protein expression system, and visualized the viral particles by transmission electron-microscopy. The in vitro-synthesized EMCV particles were indistinguishable from the in vivo-synthesized particles. Our results validate the use of the cell-free technique for the synthesis of EMCV particles.
authors.affiliation: Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo, Himeji, 671-2280, Japan. ew09k001@steng.u-hyogo.ac.jp
authors.forename: Tominari
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Jun
authors.initials: J
authors.lastname: Yukigai
authors.forename: Kosaku
authors.initials: K
authors.lastname: Ueda
authors.forename: Kodai
authors.initials: K
authors.lastname: Machida
authors.forename: Mamiko
authors.initials: M
authors.lastname: Masutani
authors.forename: Yuri
authors.initials: Y
authors.lastname: Nishino
authors.forename: Atsuo
authors.initials: A
authors.lastname: Miyazawa
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Imataka
mesh_terms: D001709:Biotechnology / Q000379:methods*; D002457:Cell Extracts / Q000302:isolation & purification*; D002474:Cell-Free System; D004686:Encephalomyocarditis virus / Q000302:isolation & purification* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D046529:Microscopy, Electron, Transmission; D014764:Viral Proteins / Q000096:biosynthesis*; D014771:Virion / Q000302:isolation & purification / Q000648:ultrastructure
chemical_list: D002457:Cell Extracts; D014764:Viral Proteins