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Generation and Comparison of Bioluminescent and Fluorescent Bacillus licheniformis

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Abstract

The environmental bacterium Bacillus licheniformis was transformed with two different shuttle-vectors (pCSS810 and pGFPratiometric) containing insect luciferase and green fluorescent protein genes, respectively. The cells were treated with various antimicrobial agents and the emitted bioluminescence and fluorescence were measured. Plasmid harboring the green fluorescent protein gene was totally segregated without selective pressure, and fluorescent B. licheniformis showed a slower growth rate than the wild-type strain; those cells were bright green as visualized by epifluorescent microscopy. However, fluorescence was not correlated to the growth state of cells or affected by the antibiotic treatments. To the contrary, luminescent transformant was found to be stable without antibiotic selection and showed analogous growth behavior compared to non-plasmid-bearing cells. The luminescent strain functioned as a biosensor for the antibiotics employed. Bioluminescence measurements allowed one to determine the viability of the recombinant cells and the kinetics of the antibacterial action could be followed. Thus, the light emission was found to be a reliable, sensitive, and real-time indicator of the “well-being” of cells, whereas fluorescence allowed one to visualize both metabolically active and inactive cells.

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Acknowledgments

We wish to thank Ville Santala for his helpful suggestions and for the laboratory assistance. We are also grateful to Katariina Tolvanen, Nina Virolainen, and Laura Nevatalo for technical support. Special thanks to Anna-Liisa Välimaa at Tampere University of Technology and Stefania Arioli at University of Milan for their kind help.

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Authors and Affiliations

  1. Department of Food Science and Microbiology, University of Milan, Via Celoria 2, 20133, Milan, Italy

    Isabella Tamagnini, Simone Guglielmetti, Diego Mora, Carlo Parini & Enrica Canzi

  2. Institute of Environmental Engineering and Biotechnology, Tampere University of Technology, 33101, Tampere, Finland

    Matti Karp

Authors
  1. Isabella Tamagnini
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  2. Simone Guglielmetti
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  3. Diego Mora
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  4. Carlo Parini
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  5. Enrica Canzi
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  6. Matti Karp
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Correspondence to Matti Karp.

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Tamagnini, I., Guglielmetti, S., Mora, D. et al. Generation and Comparison of Bioluminescent and Fluorescent Bacillus licheniformis . Curr Microbiol 57, 245–250 (2008). https://doi.org/10.1007/s00284-008-9184-y

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  • DOI: https://doi.org/10.1007/s00284-008-9184-y

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