Show simple item record

Files in this item


Item metadata

dc.contributor.authorMinskaia, Ekaterina
dc.contributor.authorRyan, Martin D.
dc.identifier.citationMinskaia , E & Ryan , M D 2013 , ' Protein coexpression using FMDV 2A : effect of “linker” residues ' , BioMed Research International .
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.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.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.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.subjectQ Scienceen
dc.titleProtein coexpression using FMDV 2A : effect of “linker” residuesen
dc.typeJournal articleen
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.description.statusPeer revieweden

This item appears in the following Collection(s)

Show simple item record