Watson, MR, Lin, Y, Hollwey, E et al. (3 more authors) (2016) An improved binary vector and Escherichia coli strain for Agrobacterium tumefaciens - mediated plant transformation. G3: Genes, Genomes, Genetics, 6 (7). pp. 2195-2201. ISSN 2160-1836
Abstract
The plasmid vector pGreenII is widely used to produce plant transformants via a process that involves propagation in Escherichia coli. However, we show here that pGreenII-based constructs can be unstable in E. coli as a consequence of them hampering cell division and promoting cell death. In addition, we describe a new version of pGreenII that does not cause these effects, thereby removing the selective pressure for mutation, and a new strain of E. coli that better tolerates existing pGreenII-based constructs without reducing plasmid yield. The adoption of the new derivative of pGreenII and E. coli strain, which we have named pViridis and MW906 respectively, should help to ensure the integrity of genes destined for study in plants, while they are propagated and manipulated in E. coli. The mechanism by which pGreenII perturbs E. coli growth appears to be dysregulation within the ColE1 origin of replication.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2016 Watson et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
Keywords: | Agrobacterium; plant transformation; pGreen; E. coli; growth defects |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biology (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 23 May 2016 10:34 |
Last Modified: | 23 Jun 2023 22:06 |
Published Version: | http://doi.org/10.1534/g3.116.029405 |
Status: | Published |
Publisher: | Genetics Society of America |
Identification Number: | 10.1534/g3.116.029405 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:100003 |