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Applied and Environmental Microbiology, July 2005, p. 3427-3432, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3427-3432.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Alternative Luciferase for Monitoring Bacterial Cells under Adverse Conditions

Siouxsie Wiles, Kathryn Ferguson, Martha Stefanidou, Douglas B. Young, and Brian D. Robertson*

Centre for Molecular Microbiology and Infection, Faculty of Medicine, Imperial College London, Flowers Building, South Kensington, London SW7 2AZ, United Kingdom

Received 22 December 2004/ Accepted 7 January 2005

The availability of cloned luciferase genes from fireflies (luc) and from bacteria (luxAB) has led to the widespread use of bioluminescence as a reporter to measure cell viability and gene expression. The most commonly occurring bioluminescence system in nature is the deep-sea imidazolopyrazine bioluminescence system. Coelenterazine is an imidazolopyrazine derivative which, when oxidized by an appropriate luciferase enzyme, produces carbon dioxide, coelenteramide, and light. The luciferase from the marine copepod Gaussia princeps (Gluc) has recently been cloned. We expressed the Gluc gene in Mycobacterium smegmatis using a shuttle vector and compared its performance with that of an existing luxAB reporter. In contrast to luxAB, the Gluc luciferase retained its luminescence output in the stationary phase of growth and exhibited enhanced stability during exposure to low pH, hydrogen peroxide, and high temperature. The work presented here demonstrated the utility of the copepod luciferase bioluminescent reporter as an alternative to bacterial luciferase, particularly for monitoring responses to environmental stress stimuli.

* Corresponding author. Mailing address: Centre for Molecular Microbiology and Infection, Faculty of Medicine, Imperial College London, London SW7 2AZ, United Kingdom. Phone: 44 020 7594 3198. Fax: 44 020 7594 3095. E-mail: b.robertson{at}imperial.ac.uk.

Applied and Environmental Microbiology, July 2005, p. 3427-3432, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3427-3432.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Bachmann, H., Santos, F., Kleerebezem, M., van Hylckama Vlieg, J. E. T. (2007). Luciferase Detection during Stationary Phase in Lactococcus lactis. Appl. Environ. Microbiol. 73: 4704-4706 [Abstract] [Full Text]  


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