A novel Gateway^®-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it>

• Main Authors: Traore Sy, Zhao Bingyu
• Format: Article
• Language: English
• Published: BMC 2011-12-01
• Series: Plant Methods
• Online Access: http://www.plantmethods.com/content/7/1/42

<p>Abstract</p> <p>Background</p> <p>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 <it>E. coli </it>strains. The ccdB protein, however, is not toxic to <it>Agrobacterium tumefaciens</it>, an important player often used for studying gene function <it>in planta</it>. This limits the direct application of the Gateway^®cloning system in plant transformation-mediated research.</p> <p>Results</p> <p>In this study, we constructed a novel Gateway^®-compatible destination vector, pEG101-SacB/R, by replacing the <it>ccdB </it>gene with a <it>SacB-SacR </it>gene cassette as the negative selectable marker.</p> <p>Conclusion</p> <p>Our results demonstrate that the new pEG101-SacB/R destination vector can be used for Gateway^®cloning in <it>Agrobacterium tumefaciens</it>. pEG101-SacB/R will be a valuable tool for high-throughput functional analysis of genes <it>in planta</it>.</p>