title: Infectious RNA transcribed in vitro from a cDNA copy of the human coronavirus genome cloned in vaccinia virus.
abstract: The coronavirus genome is a positive-strand RNA of extraordinary size and complexity. It is composed of approximately 30000 nucleotides and it is the largest known autonomously replicating RNA. It is also remarkable in that more than two-thirds of the genome is devoted to encoding proteins involved in the replication and transcription of viral RNA. Here, a reverse-genetic system is described for the generation of recombinant coronaviruses. This system is based upon the in vitro transcription of infectious RNA from a cDNA copy of the human coronavirus 229E genome that has been cloned and propagated in vaccinia virus. This system is expected to provide new insights into the molecular biology and pathogenesis of coronaviruses and to serve as a paradigm for the genetic analysis of large RNA virus genomes. It also provides a starting point for the development of a new class of eukaryotic, multi-gene RNA vectors that are able to express several proteins simultaneously.
authors.affiliation: Institute of Virology and Immunology, University of Würzburg, Versbacher Straße 7, 97078 Würzburg, Germany1.
authors.forename: Volker
authors.initials: V
authors.lastname: Thiel
authors.affiliation: Institute of Virology and Immunology, University of Würzburg, Versbacher Straße 7, 97078 Würzburg, Germany1.
authors.forename: Jens
authors.initials: J
authors.lastname: Herold
authors.affiliation: Institute of Virology and Immunology, University of Würzburg, Versbacher Straße 7, 97078 Würzburg, Germany1.
authors.forename: Barbara
authors.initials: B
authors.lastname: Schelle
authors.affiliation: Institute of Virology and Immunology, University of Würzburg, Versbacher Straße 7, 97078 Würzburg, Germany1.
authors.forename: Stuart G
authors.initials: SG
authors.lastname: Siddell
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017934:Coronavirus / Q000235:genetics* / Q000302:isolation & purification* / Q000502:physiology; D028941:Coronavirus 229E, Human; D018076:DNA, Complementary / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics; D005347:Fibroblasts; D005822:Genetic Vectors / Q000235:genetics; D016679:Genome, Viral; D000882:Haplorhini / Q000821:virology; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D014158:Transcription, Genetic / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D004274:DNA, Recombinant; D012367:RNA, Viral