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dc.contributor.authorMinskaia, Ekaterina
dc.contributor.authorRyan, Martin D.
dc.date.accessioned2013-07-23T14:31:03Z
dc.date.available2013-07-23T14:31:03Z
dc.date.issued2013-04
dc.identifier.citationMinskaia , E & Ryan , M D 2013 , ' Protein coexpression using FMDV 2A : effect of “linker” residues ' , BioMed Research International . https://doi.org/10.1155/2013/291730en
dc.identifier.issn2314-6133
dc.identifier.otherPURE: 60332211
dc.identifier.otherPURE UUID: 30268f95-4fef-42ae-9607-3c9936d3873c
dc.identifier.otherWOS: 000320794800001
dc.identifier.otherScopus: 84880154492
dc.identifier.otherORCID: /0000-0002-0012-0614/work/47136053
dc.identifier.urihttp://hdl.handle.net/10023/3864
dc.descriptionThis article was made open access through BIS OA funding. The research was supported by the MRC.en
dc.description.abstractMany biomedical applications absolutely require, or are substantially enhanced by, coexpression of multiple proteins from a single vector. Foot-and-mouth disease virus 2A (F2A) and “2A-like” sequences (e.g., Thosea asigna virus 2A; T2A) are used widely for this purpose since multiple proteins can be coexpressed by linking open reading frames (ORFs) to form a single cistron. The activity of F2A “cleavage” may, however, be compromised by both the use of shorter versions of F2A and the sequences (derived from multiple-purpose cloning sites) used to link F2A to the upstream protein. To characterise these effects, different lengths of F2A and T2A were inserted between green and cherry fluorescent proteins. Mutations were introduced in the linker region immediately upstream of both F2A- and T2A-based constructs and activities determined using both cell-free translation systems and transfected cells. In shorter versions of F2A, activity may be affected by both the C-terminal sequence of the protein upstream and, equally strikingly, the residues immediately upstream introduced during cloning. Mutations significantly improved activity for shorter versions of F2A but could decrease activity in the case of T2A. These data will aid the design of cloning strategies for the co-expression of multiple proteins in biomedical/biotechnological applications.
dc.format.extent12
dc.language.isoeng
dc.relation.ispartofBioMed Research Internationalen
dc.rightsCopyright © 2013 Ekaterina Minskaia and Martin D. Ryan. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.en
dc.subjectMultiple proteinsen
dc.subjectSingle vectoren
dc.subjectFoot-and-mouth disease virusen
dc.subjectOpen reading frames (ORFs)en
dc.subjectCistronen
dc.subjectF2Aen
dc.subjectT2Aen
dc.subjectC-terminal sequenceen
dc.subjectCloning strategiesen
dc.subjectBiomedical applicationsen
dc.subjectBiotechnological applicationsen
dc.subjectMouth-disease virusen
dc.subjectRibosome entry sitesen
dc.subjectEmbryonic stem-cellsen
dc.subjectOpen reading frameen
dc.subjectRetroviral vectoren
dc.subjectSignal sequencesen
dc.subjectGene-expressionen
dc.subjectT-cellsen
dc.subjectCleavageen
dc.subjectPeptideen
dc.subjectQ Scienceen
dc.subject.lccQen
dc.titleProtein coexpression using FMDV 2A : effect of “linker” residuesen
dc.typeJournal articleen
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/10.1155/2013/291730
dc.description.statusPeer revieweden


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