A mini-ISY100 transposon delivery system effective in γ proteobacteria

Conte, E., Mende, L., Grainge, I. and Colloms, S. D. (2019) A mini-ISY100 transposon delivery system effective in γ proteobacteria. Frontiers in Microbiology, 10, 280. (doi:10.3389/fmicb.2019.00280) (PMID:30873132) (PMCID:PMC6400869)

Conte, E., Mende, L., Grainge, I. and Colloms, S. D. (2019) A mini-ISY100 transposon delivery system effective in γ proteobacteria. Frontiers in Microbiology, 10, 280. (doi:10.3389/fmicb.2019.00280) (PMID:30873132) (PMCID:PMC6400869)

[img]
Preview
Text
179290.pdf - Published Version
Available under License Creative Commons Attribution.

3MB

Abstract

Transposons are invaluable biological tools for the genetic manipulation of microorganisms. ISY100 from Synechocystis sp. PCC6803 is a member of the Tc1/mariner/IS630 superfamily, and is characterized by high transposition efficiency and a strong preference for TA target sequences. In this paper, we describe the design and application of a mini-ISY100 suicide vector for the in vivo creation of stable random transposon insertion libraries. The system was successfully applied in seven species belonging to four different orders of γ proteobacteria. In all cases, delivery using conjugation consistently showed the highest transposition efficiency compared to chemical transformation or electroporation. We determined the frequency of transposon insertions in all the species and proved the utility of the system by identifying genes involved in colony coloration in Shewanella oneidensis. The ease and the efficiency of the protocol developed here allow the creation of complete knock-out libraries in an extensive range of host microorganisms in less than a week with no requirement for preparatory modification.

Item Type:Articles
Additional Information:This work was funded by a Leverhulme Trust grant (RP2013- K-017) to SC.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Conte, Dr Emanuele and Colloms, Dr Sean
Authors: Conte, E., Mende, L., Grainge, I., and Colloms, S. D.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Frontiers in Microbiology
Publisher:Frontiers Media
ISSN:1664-302X
ISSN (Online):1664-302X
Copyright Holders:Copyright © 2019 Conte, Mende, Grainge and Colloms
First Published:First published in Frontiers in Microbiology 10: 280
Publisher Policy:Reproduced under a Creative Commons License

University Staff: Request a correction | Enlighten Editors: Update this record