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

Blanchard, A.M. ORCID: 0000-0001-6991-7210, Egan, S.A., Emes, R.D., Warry, A. and 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. ISSN 1664-302X

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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.

Item Type: Journal article
Publication Title: Frontiers in Microbiology
Creators: Blanchard, A.M., Egan, S.A., Emes, R.D., Warry, A. and Leigh, J.A.
Publisher: Frontiers Research Foundation
Date: 2016
Volume: 7
ISSN: 1664-302X
Identifiers:
NumberType
10.3389/fmicb.2016.01645DOI
Rights: Copyright © 2016 Blanchard, Egan, Emes, Warry and Leigh. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Divisions: Schools > School of Animal, Rural and Environmental Sciences
Depositing User: Linda Sullivan
Date Added: 16 Aug 2018 10:52
Last Modified: 16 Aug 2018 11:18
URI: http://irep.ntu.ac.uk/id/eprint/34341

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