Specific monitoring by PCR amplification and bioluminescence of firefly luciferase gene-tagged bacteria added to environmental samples |
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| Authors: | Annelie Mö ller,Kersti Gustafsson,Janet K. Jansson |
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| Affiliation: | Department of Biochemistry, Arrhenius Laboratories, Stockholm University, S-10691 Stockholm, Sweden;Swedish National Chemicals Inspectorate, P.O. Box 1384, S-171 27 Solna, Sweden |
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| Abstract: | Abstract The firefly luciferase gene, luc , was demonstrated to hold promise as a specific marker for monitoring of genetically modified bacteria in the environment. PCR amplification and bioluminescence procedures were modified and compared for environmental monitoring of luc -tagged bacteria, using Escherichia coli as a model. The methods were used to track luc -tagged bacterial cells added to intact sediment core microcosms. Detection limits for the luc -tagged cells were the following, expressed as cells per 0.5 g of sediment: 102, by PCR amplification; 103, by whole cell luminescence; and 103−104, by measurement of luminescence in cell extracts. |
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| Keywords: | Firefly luciferase marker gene Bioluminescence PCR amplification Microcosms Genetically modified microorganisms |
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