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Author

Mitchell, Robert J.
Applied & Environmental Microbiology Lab (AEML)
Research Interests
  • Pathogens, bdellovibrio bacteriovorus, patho-biotechnology

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Construction and characterization of novel dual stress-responsive bacterial biosensors

Cited 27 times inthomson ciCited 31 times inthomson ci
Title
Construction and characterization of novel dual stress-responsive bacterial biosensors
Author
Mitchell, Robert J.Gu, MB
Keywords
Bacterial biosensor; Bioluminescence; Genotoxin; Green fluorescent protein; Oxidative damage; Stress
Issue Date
200404
Publisher
ELSEVIER ADVANCED TECHNOLOGY
Citation
BIOSENSORS & BIOELECTRONICS, v.19, no.9, pp.977 - 985
Abstract
Using the genes for the green fluorescence protein and Xenorhabdus luminescens luciferase operon and the promoters for the recA and katG genes, two stress-responsive Escherichia coli biosensor strains have been constructed that can individually or concurrently respond to oxidative and genotoxic conditions. Strain DUO-1 carries the pRGDK1 plasmid, which has the recA::GFPuv4 and katG::luxCDABE fusion genes oriented divergently with each other, while in DUO-2, i.e., pRGDK2, they are in a tandem orientation, with the recA promoter showing run-though transcription of the katG::luxCDABE fusion. These two strains and their responses were characterized using several known hydroxyl radical-forming chemicals, e.g., hydrogen peroxide and cadmium chloride, along with some genotoxins, e.g., mitomycin C and methyl-N-nitro-N-nitrosoguanidine, and some general toxicants. Both strains showed an induction of green fluorescent protein (GFP) and bioluminescence when they experienced DNA and oxidative damage, respectively, while the tandem orientation of the two fusion genes within DUO-2 allowed it to also sensitively respond to genotoxins via the production of bioluminescence. However, the characteristics of DUO-2's bioluminescent response to each stress were easily distinguishable, making it useful for the detection of both stresses. Furthermore, tests with mixtures of chemicals showed that both DUO-1 and DUO-2 were responsive when chemicals causing oxidative or genotoxic stress were present as a single chemical or within complex chemical mixtures.
URI
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DOI
http://dx.doi.org/10.1016/j.bios.2003.09.002
ISSN
0956-5663
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