title: Cloning full-length cDNA of grapevine chrome mosaic nepovirus.
abstract: Full-length cDNA copies of the genomic RNAs of grapevine chrome mosaic virus were obtained and cloned in Escherichia coli by a one-step procedure. The cloning protocol included size selections by agarose-gel electrophoresis of both the single-stranded and the double-stranded full-length cDNAs. First-strand cDNA synthesis was primed with oligodeoxythymidine while second-strand synthesis was primed with specific synthetic oligodeoxynucleotides, allowing cloning of the 3' poly(A) and of the last 5' nucleotides of the viral RNA template. For the 7.2-kb and 4.4-kb viral RNAs, up to 20% and 80%, respectively, of the clones were found to be full-length. Even for large templates, this procedure allows fast and efficient cloning of full-length cDNAs.
authors.affiliation: Station de Pathologie Végétale, INRA, Pont-de-la-Maye, France.
authors.forename: O
authors.initials: O
authors.lastname: Le Gall
authors.forename: T
authors.initials: T
authors.lastname: Candresse
authors.forename: V
authors.initials: V
authors.lastname: Brault
authors.forename: C
authors.initials: C
authors.lastname: Bretout
authors.forename: L
authors.initials: L
authors.lastname: Hibrand
authors.forename: J
authors.initials: J
authors.lastname: Dunez
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics
chemical_list: D004277:DNA, Single-Stranded; D004279:DNA, Viral; D012367:RNA, Viral