title: An Improved Binary Vector and Escherichia coli Strain for Agrobacterium tumefaciens-Mediated Plant Transformation.
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 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.
authors.affiliation: Centre for Plant Sciences and School of Biology, Faculty of Biological Sciences, University of Leeds, LS2 9JT, United Kingdom.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Watson
authors.affiliation: Centre for Plant Sciences and School of Biology, Faculty of Biological Sciences, University of Leeds, LS2 9JT, United Kingdom.
authors.forename: Yu-Fei
authors.initials: YF
authors.lastname: Lin
authors.affiliation: Centre for Plant Sciences and School of Biology, Faculty of Biological Sciences, University of Leeds, LS2 9JT, United Kingdom.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Hollwey
authors.affiliation: Centre for Plant Sciences and School of Biology, Faculty of Biological Sciences, University of Leeds, LS2 9JT, United Kingdom.
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Dodds
authors.affiliation: Centre for Plant Sciences and School of Biology, Faculty of Biological Sciences, University of Leeds, LS2 9JT, United Kingdom.
authors.forename: Peter
authors.initials: P
authors.lastname: Meyer
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, LS2 9JT, United Kingdom School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, LS2 9JT, United Kingdom k.j.mcdowall@leeds.ac.uk.
authors.forename: Kenneth J
authors.identifier: 0000-0003-2528-2190
authors.initials: KJ
authors.lastname: McDowall
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism / Q000382:microbiology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015169:Colony Count, Microbial; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000737:chemistry* / Q000378:metabolism; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D001426:Bacterial Proteins; C042833:ROM protein, Bacteria
keywords: Agrobacterium; E. coli; growth defects; pGreen; plant transformation