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Updated: May 26, 2026
Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector
Published on: March 28, 2018
A novel Gateway®-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens
Sy Mamadou Traore1, Bingyu Zhao
1Department of Horticulture, Virginia Polytechnic Institute and State University, Blacksburg, VA, 24061, USA. bzhao07@vt.edu.
Background:
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.
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