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Microbial genotoxicity bioreporters based on sulA activation
Alva Biran1, Hadar Ben Yoav, Sharon Yagur-Kroll
1Department of Plant and Environmental Sciences, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Israel.
Analytical and Bioanalytical Chemistry
|May 3, 2011
Summary
A novel bacterial genotoxicity reporter strain was developed using the Escherichia coli sulA promoter fused to the phoA gene. This reporter effectively detects DNA-damaging agents like hydrogen peroxide, nalidixic acid, and mitomycin C with high sensitivity.
Area of Science:
- Microbiology
- Molecular Biology
- Toxicology
Background:
- Genotoxicity testing is crucial for assessing the potential harm of chemical compounds.
- Existing methods can be time-consuming or require specialized equipment.
- Development of sensitive and rapid bacterial genotoxicity assays is needed.
Purpose of the Study:
- To construct and characterize a bacterial genotoxicity reporter strain.
- To evaluate the reporter's sensitivity and response time to DNA-damaging agents.
- To enhance reporter sensitivity through genetic modifications and integration with a detection platform.
Main Methods:
- Construction of a bacterial reporter strain by fusing the Escherichia coli sulA promoter to the phoA reporter gene.
- Testing the reporter's dose-dependent response to hydrogen peroxide, nalidixic acid (NA), and mitomycin C (MMC).
- Integration of the reporter into an electrochemical detection platform and introduction of rfaE, umuD, and uvrA knockout mutations.
Main Results:
- The reporter demonstrated a dose-dependent response to NA, MMC, and hydrogen peroxide within 30-60 minutes.
- Detection thresholds were established for MMC (0.15 μM), NA (7.5 μM), and hydrogen peroxide (~50 μM).
- Mutations in rfaE and uvrA significantly enhanced reporter sensitivity to specific genotoxic agents, including with metabolic activation.
Conclusions:
- The developed bacterial genotoxicity reporter strain offers a sensitive and rapid method for detecting DNA-damaging agents.
- Genetic modifications and platform integration further improve detection capabilities.
- This reporter system holds promise for efficient genotoxicity screening in various applications.
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