Skip to main content

Research Repository

Advanced Search

PIMMS (Pragmatic Insertional Mutation Mapping System) laboratory methodology a readily accessible tool for identification of essential genes in Streptococcus

Blanchard, Adam M.; Egan, Sharon A.; Emes, Richard D.; Warry, Andrew; Leigh, James A.

Authors

RICHARD EMES richard.emes@nottingham.ac.uk
Professor of Bioinformatics

Andrew Warry



Abstract

The Pragmatic Insertional Mutation Mapping (PIMMS) laboratory protocol was developed alongside various bioinformatics packages (Blanchard et al., 2015) to enable detection of essential and conditionally essential genes in Streptococcus and related bacteria. This extended the methodology commonly used to locate insertional mutations in individual mutants to the analysis of mutations in populations of bacteria. In Streptococcus uberis, a pyogenic Streptococcus associated with intramammary infection and mastitis in ruminants, the mutagen pGhost9:ISS1 was shown to integrate across the entire genome. Analysis of >80,000 mutations revealed 196 coding sequences, which were not be mutated and a further 67 where mutation only occurred beyond the 90th percentile of the coding sequence. These sequences showed good concordance with sequences within the database of essential genes and typically matched sequences known to be associated with basic cellular functions. Due to the broad utility of this mutagen and the simplicity of the methodology it is anticipated that PIMMS will be of value to a wide range of laboratories in functional genomic analysis of a wide range of Gram positive bacteria (Streptococcus, Enterococcus, and Lactococcus) of medical, veterinary, and industrial significance.

Citation

Blanchard, A. M., Egan, S. A., Emes, R. D., Warry, A., & Leigh, J. A. (2016). PIMMS (Pragmatic Insertional Mutation Mapping System) laboratory methodology a readily accessible tool for identification of essential genes in Streptococcus. Frontiers in Microbiology, 7(1645), https://doi.org/10.3389/fmicb.2016.01645

Journal Article Type Article
Acceptance Date Oct 3, 2016
Online Publication Date Oct 25, 2016
Publication Date Oct 25, 2016
Deposit Date Oct 26, 2016
Publicly Available Date Oct 26, 2016
Journal Frontiers in Microbiology
Electronic ISSN 1664-302X
Publisher Frontiers Media
Peer Reviewed Peer Reviewed
Volume 7
Issue 1645
DOI https://doi.org/10.3389/fmicb.2016.01645
Keywords mutagenesis, insertion sequencing, essential genome, Streptococcus, laboratory protocol
Public URL http://eprints.nottingham.ac.uk/id/eprint/37925
Publisher URL http://journal.frontiersin.org/article/10.3389/fmicb.2016.01645/full
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0
Additional Information This Document is Protected by copyright and was first published by Frontiers. All rights reserved. it is reproduced with permission.

Files






You might also like



Downloadable Citations