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

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dc.contributor.authorCummins, Joanne
dc.contributor.authorCasey, Pat G.
dc.contributor.authorJoyce, Susan A.
dc.contributor.authorGahan, Cormac G.en
dc.date.accessioned2016-02-17T11:45:32Z
dc.date.available2016-02-17T11:45:32Z
dc.date.issued2013
dc.description.abstractListeria 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.sponsorshipScience Foundation Ireland (08/RFP/GEN1320)en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.articleide75437
dc.identifier.citationCummins 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.doi10.1371/journal.pone.0075437
dc.identifier.issn1932-6203
dc.identifier.issued9en
dc.identifier.journaltitlePLOS ONEen
dc.identifier.urihttps://hdl.handle.net/10468/2365
dc.identifier.volume8en
dc.language.isoenen
dc.publisherPublic Library of Scienceen
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 crediteden
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectProtoporphyrinogen oxidaseen
dc.subjectGastrointestinal phaseen
dc.subjectCrystal structureen
dc.subjectEscherichia colien
dc.subjectVirulence genesen
dc.subjectCrp/Fnr familyen
dc.subjectIdentificationen
dc.subjectGenomeen
dc.subjectInternalinen
dc.subjectCellsen
dc.titleA mariner transposon-based signature-tagged mutagenesis system for the analysis of oral infection by Listeria monocytogenesen
dc.typeArticle (peer-reviewed)en
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