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A novel Gateway(®)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens
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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3265438/ https://www.ncbi.nlm.nih.gov/pubmed/22145613 http://dx.doi.org/10.1186/1746-4811-7-42 |
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author | Traore, Sy Mamadou Zhao, Bingyu |
author_facet | Traore, Sy Mamadou Zhao, Bingyu |
author_sort | Traore, Sy Mamadou |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3265438 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-32654382012-01-25 A novel Gateway(®)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens Traore, Sy Mamadou Zhao, Bingyu Plant Methods Methodology 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. BioMed Central 2011-12-07 /pmc/articles/PMC3265438/ /pubmed/22145613 http://dx.doi.org/10.1186/1746-4811-7-42 Text en Copyright ©2011 Traore and Zhao; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Traore, Sy Mamadou Zhao, Bingyu A novel Gateway(®)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens |
title | A novel Gateway(®)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens |
title_full | A novel Gateway(®)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens |
title_fullStr | A novel Gateway(®)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens |
title_full_unstemmed | A novel Gateway(®)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens |
title_short | A novel Gateway(®)-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens |
title_sort | novel gateway(®)-compatible binary vector allows direct selection of recombinant clones in agrobacterium tumefaciens |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3265438/ https://www.ncbi.nlm.nih.gov/pubmed/22145613 http://dx.doi.org/10.1186/1746-4811-7-42 |
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