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dc.contributor.authorBrennan, Benjamin
dc.contributor.authorLi, Ping
dc.contributor.authorElliott, Richard M.
dc.date.accessioned2012-01-20T13:01:02Z
dc.date.available2012-01-20T13:01:02Z
dc.date.issued2011-12
dc.identifier.citationBrennan , B , Li , P & Elliott , R M 2011 , ' Generation and characterization of a recombinant Rift Valley fever virus expressing a V5 epitope-tagged RNA-dependent RNA polymerase ' , Journal of General Virology , vol. 92 , no. 12 , pp. 2906-2913 . https://doi.org/10.1099/vir.0.036749-0en
dc.identifier.issn0022-1317
dc.identifier.otherPURE: 16601767
dc.identifier.otherPURE UUID: e79cd4b0-a39d-4bdf-8622-819e078ca654
dc.identifier.otherWOS: 000298207700024
dc.identifier.otherScopus: 81255143065
dc.identifier.urihttps://hdl.handle.net/10023/2191
dc.description.abstractThe viral RNA-dependent RNA polymerase (RdRp; L protein) of Rift Valley fever virus (RVFV; family Bunyaviridae) is a 238 kDa protein that is crucial for the life cycle of the virus, as it catalyses both transcription of viral mRNAs and replication of the tripartite genome. Despite its importance, little is known about the intracellular distribution of the polymerase or its other roles during infection, primarily because of lack of specific antibodies that recognize L protein. To begin to address these questions we investigated whether the RVFV (MP12 strain) polymerase could tolerate insertion of the V5 epitope, as has been previously demonstrated for the Bunyamwera virus L protein. Insertion of the 14 aa epitope into the polymerase sequence at aa 1852 resulted in a polymerase that retained functionality in a minigenome assay, and we were able to rescue recombinant viruses that expressed the modified L protein by reverse genetics. The L protein could be detected in infected cells by Western blotting with anti-V5 antibodies. Examination of recombinant virus-infected cells by immunofluorescence revealed a punctate perinuclear or cytoplasmic distribution of the polymerase that co-localized with the nucleocapsid protein. The generation of RVFV expressing a tagged RdRp will allow detailed examination of the role of the viral polymerase in the virus life cycle.
dc.format.extent8
dc.language.isoeng
dc.relation.ispartofJournal of General Virologyen
dc.rights(c) 2011 SGM. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.en
dc.subjectViral-RNAen
dc.subjectL-proteinen
dc.subjectBunyamwera virusen
dc.subjectL-segmenten
dc.subjectReplicationen
dc.subjectRescueen
dc.subjectGenomeen
dc.subjectTranscriptionen
dc.subjectSequenceen
dc.subjectidentificationen
dc.subjectQR355 Virologyen
dc.subject.lccQR355en
dc.titleGeneration and characterization of a recombinant Rift Valley fever virus expressing a V5 epitope-tagged RNA-dependent RNA polymeraseen
dc.typeJournal articleen
dc.contributor.sponsorBBSRCen
dc.contributor.sponsorEuropean Commissionen
dc.contributor.sponsorThe Wellcome Trusten
dc.contributor.sponsorMedical Research Councilen
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews. School of Biologyen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.identifier.doihttps://doi.org/10.1099/vir.0.036749-0
dc.description.statusPeer revieweden
dc.identifier.grantnumberBB/G004277/1en
dc.identifier.grantnumber211757en
dc.identifier.grantnumber079810/Z/06/Zen
dc.identifier.grantnumberG0800161en


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