A novel Gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it>
<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<sup>® </sup>cloning system based on the site-specific DNA recombination propertie...
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doaj-1fe3f061d7984f7494e71370f70b06842020-11-24T23:34:32ZengBMCPlant Methods1746-48112011-12-01714210.1186/1746-4811-7-42A novel Gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it>Traore SyZhao Bingyu<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<sup>® </sup>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<sup>® </sup>cloning system in plant transformation-mediated research.</p> <p>Results</p> <p>In this study, we constructed a novel Gateway<sup>®</sup>-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<sup>® </sup>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> http://www.plantmethods.com/content/7/1/42 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Traore Sy Zhao Bingyu |
spellingShingle |
Traore Sy Zhao Bingyu A novel Gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it> Plant Methods |
author_facet |
Traore Sy Zhao Bingyu |
author_sort |
Traore Sy |
title |
A novel Gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it> |
title_short |
A novel Gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it> |
title_full |
A novel Gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it> |
title_fullStr |
A novel Gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it> |
title_full_unstemmed |
A novel Gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it> |
title_sort |
novel gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>agrobacterium tumefaciens</it> |
publisher |
BMC |
series |
Plant Methods |
issn |
1746-4811 |
publishDate |
2011-12-01 |
description |
<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<sup>® </sup>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<sup>® </sup>cloning system in plant transformation-mediated research.</p> <p>Results</p> <p>In this study, we constructed a novel Gateway<sup>®</sup>-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<sup>® </sup>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> |
url |
http://www.plantmethods.com/content/7/1/42 |
work_keys_str_mv |
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