A mariner transposon-based signature-tagged mutagenesis system for the analysis of oral infection by Listeria monocytogenes

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dc.contributor.author Cummins, Joanne
dc.contributor.author Casey, Pat G.
dc.contributor.author Joyce, Susan A.
dc.contributor.author Gahan, Cormac G. en
dc.date.accessioned 2016-02-17T11:45:32Z
dc.date.available 2016-02-17T11:45:32Z
dc.date.issued 2013
dc.identifier.citation Cummins J, Casey PG, Joyce SA, Gahan CGM (2013) A Mariner Transposon-Based Signature-Tagged Mutagenesis System for the Analysis of Oral Infection by Listeria monocytogenes. PLoS ONE 8(9): e75437. doi:10.1371/journal.pone.0075437
dc.identifier.volume 8 en
dc.identifier.issued 9 en
dc.identifier.issn 1932-6203
dc.identifier.uri http://hdl.handle.net/10468/2365
dc.identifier.doi 10.1371/journal.pone.0075437
dc.description.abstract Listeria monocytogenes is a Gram-positive foodborne pathogen and the causative agent of listerosis a disease that manifests predominately as meningitis in the non-pregnant individual or infection of the fetus and spontaneous abortion in pregnant women. Common-source outbreaks of foodborne listeriosis are associated with significant morbidity and mortality. However, relatively little is known concerning the mechanisms that govern infection via the oral route. In order to aid functional genetic analysis of the gastrointestinal phase of infection we designed a novel signature-tagged mutagenesis (STM) system based upon the invasive L. monocytogenes 4b serotype H7858 strain. To overcome the limitations of gastrointestinal infection by L. monocytogenes in the mouse model we created a H7858 strain that is genetically optimised for oral infection in mice. Furthermore our STM system was based upon a mariner transposon to favour numerous and random transposition events throughout the L. monocytogenes genome. Use of the STM bank to investigate oral infection by L. monocytogenes identified 21 insertion mutants that demonstrated significantly reduced potential for infection in our model. The sites of transposon insertion included lmOh7858_0671 (encoding an internalin homologous to Lmo0610), lmOh7858_0898 (encoding a putative surface-expressed LPXTG protein homologous to Lmo0842), lmOh7858_2579 (encoding the HupDGC hemin transport system) and lmOh7858_0399 (encoding a putative fructose specific phosphotransferase system). We propose that this represents an optimised STM system for functional genetic analysis of foodborne/oral infection by L. monocytogenes. en
dc.description.sponsorship Science Foundation Ireland (08/RFP/GEN1320) en
dc.format.mimetype application/pdf en
dc.language.iso en en
dc.publisher Public Library of Science en
dc.rights © 2015 Cummins et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited en
dc.rights.uri http://creativecommons.org/licenses/by/4.0/ en
dc.subject Protoporphyrinogen oxidase en
dc.subject Gastrointestinal phase en
dc.subject Crystal structure en
dc.subject Escherichia coli en
dc.subject Virulence genes en
dc.subject Crp/Fnr family en
dc.subject Identification en
dc.subject Genome en
dc.subject Internalin en
dc.subject Cells en
dc.title A mariner transposon-based signature-tagged mutagenesis system for the analysis of oral infection by Listeria monocytogenes en
dc.type Article (peer-reviewed) en
dc.internal.authorcontactother Cormac Gahan, Microbiology, University College Cork, Cork, Ireland. +353-21-490-3000 Email: c.gahan@ucc.ie en
dc.internal.availability Full text available en
dc.description.version Published Version en
dc.internal.rssid 329812465
dc.internal.wokid WOS:000326240100125
dc.description.status Peer reviewed en
dc.identifier.journaltitle PLOS ONE en
dc.internal.IRISemailaddress c.gahan@ucc.ie en
dc.internal.IRISemailaddress s.joyce@ucc.ie en
dc.identifier.articleid e75437


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© 2015 Cummins et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Except where otherwise noted, this item's license is described as © 2015 Cummins et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
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