A novel Gateway(R)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens

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dc.contributor.authorTraore, Sy M.en
dc.contributor.authorZhao, Bingyu Y.en
dc.contributor.departmentSchool of Plant and Environmental Sciencesen
dc.date.accessioned2012-08-24T10:49:28Zen
dc.date.available2012-08-24T10:49:28Zen
dc.date.issued2011-12-07en
dc.date.updated2012-08-24T10:49:28Zen
dc.description.abstractBackground Cloning genes into plasmid vectors is one of the key steps for studying gene function. Recently, Invitrogen& developed a convenient Gateway® cloning system based on the site-specific DNA recombination properties of bacteriophage lambda and the cytotoxic protein ccdB, which is lethal to most E. coli strains. The ccdB protein, however, is not toxic to Agrobacterium tumefaciens, an important player often used for studying gene function in planta. This limits the direct application of the Gateway® cloning system in plant transformation-mediated research. Results In this study, we constructed a novel Gateway®-compatible destination vector, pEG101-SacB/R, by replacing the ccdB gene with a SacB-SacR gene cassette as the negative selectable marker. Conclusion Our results demonstrate that the new pEG101-SacB/R destination vector can be used for Gateway® cloning in Agrobacterium tumefaciens. pEG101-SacB/R will be a valuable tool for high-throughput functional analysis of genes in planta.en
dc.description.sponsorshipFinancial support was provided by grants from Binational Agricultural Research and Development Fund (BARD) (US-4216-09 to BZ), US NSF (IOS- 0845283 to BZ), and the Virginia Agricultural Experiment Station (VA135872).en
dc.description.versionPublished versionen
dc.format.extent8 pagesen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationPlant Methods. 2011 Dec 07;7(1):42en
dc.identifier.doihttps://doi.org/10.1186/1746-4811-7-42en
dc.identifier.urihttp://hdl.handle.net/10919/18779en
dc.language.isoenen
dc.publisherBMCen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.holderSy Traore et al.; licensee BioMed Central Ltd.en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.titleA novel Gateway(R)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciensen
dc.title.serialPlant Methodsen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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