An improved conjugation method for Pseudomonas syringae

Robert Jackson; Michelle Hulin; Helen Neale; Richard Harrison; John Mansfield; Dawn Arnold

doi:10.1016/j.mimet.2020.106025

An improved conjugation method for Pseudomonas syringae

Robert Jackson, Michelle Hulin, Helen Neale, Richard Harrison, John Mansfield, Dawn Arnold

Research output: Contribution to journal › Article › peer-review

Abstract

In order to achieve saturating transposon mutagenesis of the genome of plant pathogenic strains of Pseudomonas syringae we needed to improve plasmid conjugation frequency. Manipulation of the growth stage of donor and recipient cells allowed the required increase in frequency and facilitated conjugation of otherwise recalcitrant strains.

Original language	English
Article number	106025
Journal	Journal of Microbiological Methods
Volume	177
DOIs	https://doi.org/10.1016/j.mimet.2020.106025
Publication status	Published - Oct 2020

Bibliographical note

Funding Information:
We thank Adam Deutschbauer (Lawrence Berkeley National Laboratory, Berkeley, California, USA) for providing us with mariner vector pKMW3. This work was carried out under the Department for Environment, Food and Rural Affairs' Plant Health and Seeds Inspectorate license number 51049/202078/1. This research was funded by grants from the Biotechnology and Biological Sciences Research Council , United Kingdom (UWE grant number BB/P005705/1 and EMR grant number BB/P006272/1 ).

Publisher Copyright:
© 2020 Elsevier B.V.

Keywords

Conjugation
Pseudomonas
TnSeq

ASJC Scopus subject areas

Microbiology
Molecular Biology
Microbiology (medical)

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10.1016/j.mimet.2020.106025

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title = "An improved conjugation method for Pseudomonas syringae",

abstract = "In order to achieve saturating transposon mutagenesis of the genome of plant pathogenic strains of Pseudomonas syringae we needed to improve plasmid conjugation frequency. Manipulation of the growth stage of donor and recipient cells allowed the required increase in frequency and facilitated conjugation of otherwise recalcitrant strains.",

keywords = "Conjugation, Pseudomonas, TnSeq",

author = "Robert Jackson and Michelle Hulin and Helen Neale and Richard Harrison and John Mansfield and Dawn Arnold",

note = "Funding Information: We thank Adam Deutschbauer (Lawrence Berkeley National Laboratory, Berkeley, California, USA) for providing us with mariner vector pKMW3. This work was carried out under the Department for Environment, Food and Rural Affairs' Plant Health and Seeds Inspectorate license number 51049/202078/1. This research was funded by grants from the Biotechnology and Biological Sciences Research Council , United Kingdom (UWE grant number BB/P005705/1 and EMR grant number BB/P006272/1 ). Publisher Copyright: {\textcopyright} 2020 Elsevier B.V.",

year = "2020",

month = oct,

doi = "10.1016/j.mimet.2020.106025",

language = "English",

volume = "177",

journal = "Journal of Microbiological Methods",

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T1 - An improved conjugation method for Pseudomonas syringae

AU - Jackson, Robert

AU - Hulin, Michelle

AU - Neale, Helen

AU - Harrison, Richard

AU - Mansfield, John

AU - Arnold, Dawn

N1 - Funding Information: We thank Adam Deutschbauer (Lawrence Berkeley National Laboratory, Berkeley, California, USA) for providing us with mariner vector pKMW3. This work was carried out under the Department for Environment, Food and Rural Affairs' Plant Health and Seeds Inspectorate license number 51049/202078/1. This research was funded by grants from the Biotechnology and Biological Sciences Research Council , United Kingdom (UWE grant number BB/P005705/1 and EMR grant number BB/P006272/1 ). Publisher Copyright: © 2020 Elsevier B.V.

PY - 2020/10

Y1 - 2020/10

N2 - In order to achieve saturating transposon mutagenesis of the genome of plant pathogenic strains of Pseudomonas syringae we needed to improve plasmid conjugation frequency. Manipulation of the growth stage of donor and recipient cells allowed the required increase in frequency and facilitated conjugation of otherwise recalcitrant strains.

AB - In order to achieve saturating transposon mutagenesis of the genome of plant pathogenic strains of Pseudomonas syringae we needed to improve plasmid conjugation frequency. Manipulation of the growth stage of donor and recipient cells allowed the required increase in frequency and facilitated conjugation of otherwise recalcitrant strains.

KW - Conjugation

KW - Pseudomonas

KW - TnSeq

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U2 - 10.1016/j.mimet.2020.106025

DO - 10.1016/j.mimet.2020.106025

M3 - Article

SN - 0167-7012

VL - 177

JO - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

M1 - 106025

ER -

An improved conjugation method for Pseudomonas syringae

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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