Alternative luciferase for monitoring bacterial cells under adverse conditions.
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.
Wiles
Siouxsie
S
Centre for Molecular Microbiology and Infection, Faculty of Medicine, Imperial College London, London SW7 2AZ, United Kingdom.
Ferguson
Kathryn
K
Stefanidou
Martha
M
Young
Douglas B
DB
Robertson
Brian D
BD
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015169:Colony Count, Microbial; D033342:Copepoda / Q000201:enzymology* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D018869:Heat-Shock Response; D006861:Hydrogen Peroxide; D006863:Hydrogen-Ion Concentration; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements; D020102:Mycobacterium smegmatis / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000502:physiology*; D013696:Temperature
D001426:Bacterial Proteins; D006861:Hydrogen Peroxide; D008156:Luciferases