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...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2011-12-01
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| Series: | Plant Methods |
| Online Access: | http://www.plantmethods.com/content/7/1/42 |
| _version_ | 1818515972971036672 |
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| author | Traore Sy Zhao Bingyu |
| author_facet | Traore Sy Zhao Bingyu |
| author_sort | Traore Sy |
| collection | DOAJ |
| 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> |
| first_indexed | 2024-12-11T00:35:56Z |
| format | Article |
| id | doaj.art-1fe3f061d7984f7494e71370f70b0684 |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-12-11T00:35:56Z |
| publishDate | 2011-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-1fe3f061d7984f7494e71370f70b06842022-12-22T01:27:08ZengBMCPlant 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 |
| 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 |
| title | 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_short | A novel Gateway<sup>®</sup>-compatible binary vector allows direct selection of recombinant clones in <it>Agrobacterium tumefaciens</it> |
| title_sort | novel gateway sup r sup compatible binary vector allows direct selection of recombinant clones in it agrobacterium tumefaciens it |
| url | http://www.plantmethods.com/content/7/1/42 |
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