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A mariner transposon-based signature-tagged mutagenesis system for the analysis of oral infection by Listeria monocytogenes.
PLoS One. 2013; 8(9):e75437.Plos

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.

Authors+Show Affiliations

Department of Microbiology, University College Cork, Cork, Ireland.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24069416

Citation

Cummins, Joanne, et al. "A Mariner Transposon-based Signature-tagged Mutagenesis System for the Analysis of Oral Infection By Listeria Monocytogenes." PloS One, vol. 8, no. 9, 2013, pp. e75437.
Cummins J, Casey PG, Joyce SA, et al. A mariner transposon-based signature-tagged mutagenesis system for the analysis of oral infection by Listeria monocytogenes. PLoS ONE. 2013;8(9):e75437.
Cummins, J., Casey, P. G., Joyce, S. A., & Gahan, C. G. (2013). A mariner transposon-based signature-tagged mutagenesis system for the analysis of oral infection by Listeria monocytogenes. PloS One, 8(9), e75437. https://doi.org/10.1371/journal.pone.0075437
Cummins J, et al. A Mariner Transposon-based Signature-tagged Mutagenesis System for the Analysis of Oral Infection By Listeria Monocytogenes. PLoS ONE. 2013;8(9):e75437. PubMed PMID: 24069416.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A mariner transposon-based signature-tagged mutagenesis system for the analysis of oral infection by Listeria monocytogenes. AU - Cummins,Joanne, AU - Casey,Pat G, AU - Joyce,Susan A, AU - Gahan,Cormac G M, Y1 - 2013/09/12/ PY - 2013/03/13/received PY - 2013/08/14/accepted PY - 2013/9/27/entrez PY - 2013/9/27/pubmed PY - 2014/6/16/medline SP - e75437 EP - e75437 JF - PloS one JO - PLoS ONE VL - 8 IS - 9 N2 - 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. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/24069416/A_mariner_transposon-based_signature-tagged_mutagenesis_system_for_the_analysis_of_oral_infection_by_Listeria_monocytogenes L2 - http://dx.plos.org/10.1371/journal.pone.0075437 DB - PRIME DP - Unbound Medicine ER -