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Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon

BACKGROUND: Salmonellosis may be a food safety problem when raw food products are mishandled and not fully cooked. In previous work, we developed bioluminescent Salmonella enterica serotypes using a plasmid-based reporting system that can be used for real-time monitoring of the pathogen's growt...

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Autores principales: Howe, Kevin, Karsi, Attila, Germon, Pierre, Wills, Robert W, Lawrence, Mark L, Bailey, Richard H
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2918591/
https://www.ncbi.nlm.nih.gov/pubmed/20653968
http://dx.doi.org/10.1186/1471-2180-10-197
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author Howe, Kevin
Karsi, Attila
Germon, Pierre
Wills, Robert W
Lawrence, Mark L
Bailey, Richard H
author_facet Howe, Kevin
Karsi, Attila
Germon, Pierre
Wills, Robert W
Lawrence, Mark L
Bailey, Richard H
author_sort Howe, Kevin
collection PubMed
description BACKGROUND: Salmonellosis may be a food safety problem when raw food products are mishandled and not fully cooked. In previous work, we developed bioluminescent Salmonella enterica serotypes using a plasmid-based reporting system that can be used for real-time monitoring of the pathogen's growth on food products in short term studies. In this study, we report the use of a Tn7-based transposon system for subcloning of luxCDABE genes into the chromosome of eleven Salmonella enterica serotypes isolated from the broiler production continuum. RESULTS: We found that the lux operon is constitutively expressed from the chromosome post-transposition and the lux cassette is stable without external pressure, i.e. antibiotic selection, for all Salmonella enterica serotypes used. Bioluminescence expression is based on an active electron transport chain and is directly related with metabolic activity. This relationship was quantified by measuring bioluminescence against a temperature gradient in aqueous solution using a luminometer. In addition, bioluminescent monitoring of two serotypes confirmed that our chicken skin model has the potential to be used to evaluate pathogen mitigation strategies. CONCLUSIONS: This study demonstrated that our new stable reporting system eliminates bioluminescence variation due to plasmid instability and provides a reliable real-time experimental system to study application of preventive measures for Salmonella on food products in real-time for both short and long term studies.
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spelling pubmed-29185912010-08-10 Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon Howe, Kevin Karsi, Attila Germon, Pierre Wills, Robert W Lawrence, Mark L Bailey, Richard H BMC Microbiol Research Article BACKGROUND: Salmonellosis may be a food safety problem when raw food products are mishandled and not fully cooked. In previous work, we developed bioluminescent Salmonella enterica serotypes using a plasmid-based reporting system that can be used for real-time monitoring of the pathogen's growth on food products in short term studies. In this study, we report the use of a Tn7-based transposon system for subcloning of luxCDABE genes into the chromosome of eleven Salmonella enterica serotypes isolated from the broiler production continuum. RESULTS: We found that the lux operon is constitutively expressed from the chromosome post-transposition and the lux cassette is stable without external pressure, i.e. antibiotic selection, for all Salmonella enterica serotypes used. Bioluminescence expression is based on an active electron transport chain and is directly related with metabolic activity. This relationship was quantified by measuring bioluminescence against a temperature gradient in aqueous solution using a luminometer. In addition, bioluminescent monitoring of two serotypes confirmed that our chicken skin model has the potential to be used to evaluate pathogen mitigation strategies. CONCLUSIONS: This study demonstrated that our new stable reporting system eliminates bioluminescence variation due to plasmid instability and provides a reliable real-time experimental system to study application of preventive measures for Salmonella on food products in real-time for both short and long term studies. BioMed Central 2010-07-23 /pmc/articles/PMC2918591/ /pubmed/20653968 http://dx.doi.org/10.1186/1471-2180-10-197 Text en Copyright ©2010 Howe et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Howe, Kevin
Karsi, Attila
Germon, Pierre
Wills, Robert W
Lawrence, Mark L
Bailey, Richard H
Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon
title Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon
title_full Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon
title_fullStr Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon
title_full_unstemmed Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon
title_short Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon
title_sort development of stable reporter system cloning luxcdabe genes into chromosome of salmonella enterica serotypes using tn7 transposon
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2918591/
https://www.ncbi.nlm.nih.gov/pubmed/20653968
http://dx.doi.org/10.1186/1471-2180-10-197
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