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dc.contributor.authorOsman, T. A. M.
dc.contributor.authorMorris, J.
dc.contributor.authorCoutts, Robert H.A.
dc.contributor.authorBuck, K. W.
dc.identifier.citationOsman , T A M , Morris , J , Coutts , R H A & Buck , K W 2006 , ' Synthesis of genomic and subgenomic RNAs by a membrane-bound RNA-dependent RNA polymerase isolated from oat plants infected with cereal yellow dwarf virus ' , Archives of Virology , vol. 151 , no. 11 , pp. 2229-2242 .
dc.identifier.otherPURE: 1953587
dc.identifier.otherPURE UUID: 981831f1-0956-4654-b33b-ac5f470ec226
dc.identifier.otherWOS: 000241452800010
dc.identifier.otherScopus: 33750314157
dc.description.abstractA membrane-bound RNA-dependent RNA polymerase (RdRp) complex was isolated by differential sedimentation from oat plants infected with cereal yellow dwarf virus (CYDV). When incubated with P-32-labelled UTP, unlabelled ATP, CTP and GTP, and Mg2+ ions, the RdRp preparation catalysed the synthesis of double-stranded (ds) RNAs corresponding in size to the virus genomic RNA (5.7 kbp) and two putative subgenomic RNAs (2.8 and 0.7 kbp). Hybridisation using strand-specific hybridization targets showed that the 5.7-kbp dsRNA was labelled mainly in the plus strand, whereas the 2.8- and 0.7-kbp dsRNAs were labelled only in the minus strand. Genomic-length single-stranded, plus-strand RNA of 5.7 kb and single-stranded, plus-strand subgenomic RNAs of 2.8 and 0.7 kbp were detected in RNA isolated from oat plants infected with CYDV. Mapping experiments were consistent with the genomic and subgenomic RNAs having common 3' ends, but different 5' ends, whether produced in vitro or in vivo. The RdRp-encoding region of the CYDV genome was cloned and expressed in Escherichia coli, and the purified protein was used to raise antibodies in a rabbit. In immunoblots, the antibodies detected a protein of about 68 kDa in RdRp preparations from CYDV-infected oat plants, but not from equivalent preparations from healthy oats. As far as we are aware, this is the first report of an in vitro RNA synthesis system for a phloem-limited virus.en
dc.relation.ispartofArchives of Virology
dc.titleSynthesis of genomic and subgenomic RNAs by a membrane-bound RNA-dependent RNA polymerase isolated from oat plants infected with cereal yellow dwarf virusen
dc.contributor.institutionGeography, Environment and Agriculture
dc.contributor.institutionAgriculture, Veterinary and Food Sciences
dc.contributor.institutionCrop and Environmental Protection
dc.contributor.institutionSchool of Life and Medical Sciences
dc.contributor.institutionHealth & Human Sciences Research Institute
dc.contributor.institutionDepartment of Human and Environmental Sciences
dc.description.statusPeer reviewed
dc.relation.schoolSchool of Life and Medical Sciences
rioxxterms.typeJournal Article/Review

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