title: Human cell extract-derived cell-free systems for virus synthesis.
abstract: Cell-free synthesis of an infectious virus is an ideal tool for elucidating the mechanism of viral replication and for development of antiviral drugs. In this chapter, the synthesis of Encephalomyocarditis virus (EMCV) from RNA and DNA in a HeLa cell extract-derived in vitro protein expression system is described. When a synthetic EMCV RNA with a hammerhead ribozyme sequence at its 5'-end is incubated with a HeLa cell extract using a dialysis system, EMCV particles are progressively synthesized. For EMCV synthesis from DNA, a plasmid harboring the full-length cDNA of EMCV with the T7 promoter/terminator unit is incubated in the HeLa cell extract supplemented with T7 RNA polymerase.
authors.affiliation: Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo, Himeji, Japan.
authors.forename: Tominari
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Kodai
authors.initials: K
authors.lastname: Machida
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Imataka
mesh_terms: D018929:Cell Culture Techniques; D002457:Cell Extracts; D002474:Cell-Free System; D004279:DNA, Viral / Q000235:genetics; D004686:Encephalomyocarditis virus / Q000235:genetics* / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids / Q000235:genetics; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D013698:Templates, Genetic; D014158:Transcription, Genetic
chemical_list: D002457:Cell Extracts; D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral