A flow cytometry-optimized assay using an SOS-green fluorescent protein (SOS-GFP) whole-cell biosensor for the detection of genotoxins in complex environments.
Whole-cell biosensors have become popular tools for detection of ecotoxic compounds in environmental samples. We have developed an assay optimized for flow cytometry with detection of genotoxic compounds in mind. The assay features extended pre-incubation and a cell density of only 10(6)-10(7) cells/mL, and proved far more sensitive than a previously published assay using the same biosensor strain. By applying the SOS-green fluorescent protein (GFP) whole-cell biosensor directly to soil microcosms we were also able to evaluate both the applicability and sensitivity of a biosensor based on SOS-induction in whole soil samples. Soil microcosms were spiked with a dilution-series of crude broth extract from the mitomycin C-producing streptomycete Streptomyces caespitosus. Biosensors extracted from these microcosms after 1 day of incubation at 30 degrees C were easily distinguished from extracts of non-contaminated soil particles when using flow cytometry, and induction of the biosensor by mitomycin C was detectable at concentrations as low as 2.5 ng/g of soil.
Norman
Anders
A
Department of Microbiology, University of Copenhagen, 1307 Copenhagen K, Denmark.
Hansen
Lars Hestbjerg
LH
Sørensen
Søren J
SJ
D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D015374:Biosensing Techniques; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins; D008769:Methylnitronitrosoguanidine / Q000633:toxicity; D016685:Mitomycin / Q000378:metabolism*; D013014:SOS Response, Genetics; D012988:Soil Microbiology; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*
D000903:Antibiotics, Antineoplastic; D008769:Methylnitronitrosoguanidine; D049452:Green Fluorescent Proteins; D016685:Mitomycin