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An Improved Binary Vector and Escherichia coli Strain for Agrobacterium tumefaciens-Mediated Plant Transformation

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 additio...

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Autores principales: Watson, Michael R., Lin, Yu-fei, Hollwey, Elizabeth, Dodds, Rachel E., Meyer, Peter, McDowall, Kenneth J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Genetics Society of America 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4938672/
https://www.ncbi.nlm.nih.gov/pubmed/27194805
http://dx.doi.org/10.1534/g3.116.029405
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author Watson, Michael R.
Lin, Yu-fei
Hollwey, Elizabeth
Dodds, Rachel E.
Meyer, Peter
McDowall, Kenneth J.
author_facet Watson, Michael R.
Lin, Yu-fei
Hollwey, Elizabeth
Dodds, Rachel E.
Meyer, Peter
McDowall, Kenneth J.
author_sort Watson, Michael R.
collection PubMed
description 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 the 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.
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spelling pubmed-49386722016-07-19 An Improved Binary Vector and Escherichia coli Strain for Agrobacterium tumefaciens-Mediated Plant Transformation Watson, Michael R. Lin, Yu-fei Hollwey, Elizabeth Dodds, Rachel E. Meyer, Peter McDowall, Kenneth J. G3 (Bethesda) Investigations 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 the 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. Genetics Society of America 2016-05-17 /pmc/articles/PMC4938672/ /pubmed/27194805 http://dx.doi.org/10.1534/g3.116.029405 Text en Copyright © 2016 Watson et al. http://creativecommons.org/licenses/by/4.0/ 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.
spellingShingle Investigations
Watson, Michael R.
Lin, Yu-fei
Hollwey, Elizabeth
Dodds, Rachel E.
Meyer, Peter
McDowall, Kenneth J.
An Improved Binary Vector and Escherichia coli Strain for Agrobacterium tumefaciens-Mediated Plant Transformation
title An Improved Binary Vector and Escherichia coli Strain for Agrobacterium tumefaciens-Mediated Plant Transformation
title_full An Improved Binary Vector and Escherichia coli Strain for Agrobacterium tumefaciens-Mediated Plant Transformation
title_fullStr An Improved Binary Vector and Escherichia coli Strain for Agrobacterium tumefaciens-Mediated Plant Transformation
title_full_unstemmed An Improved Binary Vector and Escherichia coli Strain for Agrobacterium tumefaciens-Mediated Plant Transformation
title_short An Improved Binary Vector and Escherichia coli Strain for Agrobacterium tumefaciens-Mediated Plant Transformation
title_sort improved binary vector and escherichia coli strain for agrobacterium tumefaciens-mediated plant transformation
topic Investigations
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4938672/
https://www.ncbi.nlm.nih.gov/pubmed/27194805
http://dx.doi.org/10.1534/g3.116.029405
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