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

• Main Authors: Lawrence Mark L, Wills Robert W, Germon Pierre, Karsi Attila, Howe Kevin, Bailey Richard H
• Format: Article
• Language: English
• Published: BMC 2010-07-01
• Series: BMC Microbiology
• Online Access: http://www.biomedcentral.com/1471-2180/10/197

<p>Abstract</p> <p>Background</p> <p>Salmonellosis may be a food safety problem when raw food products are mishandled and not fully cooked. In previous work, we developed bioluminescent <it>Salmonella enterica </it>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 <it>luxCDABE </it>genes into the chromosome of eleven <it>Salmonella enterica </it>serotypes isolated from the broiler production continuum.</p> <p>Results</p> <p>We found that the <it>lux </it>operon is constitutively expressed from the chromosome post-transposition and the <it>lux </it>cassette is stable without external pressure, i.e. antibiotic selection, for all <it>Salmonella enterica </it>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.</p> <p>Conclusions</p> <p>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 <it>Salmonella </it>on food products in real-time for both short and long term studies.</p>