The utilization of a Saccharomyces cerevisiae HUG1P -GFP promoter–reporter construct for the selective detection of DNA damage

The utilization of a Saccharomyces cerevisiae HUG1P -GFP promoter–reporter construct for the... In this study, we report the creation and characterization of a yeast-based promoter–reporter construct for the detection of genotoxic compounds within a cell's local environment. We have synthesized a fusion containing the HUG1 promoter and GFP and incorporated this cassette into the yeast genome creating a stable, sensitive genotoxicity indicator. To quantify biosensor performance, HUG1P -GFP cells were exposed to multiple doses of a wide variety of genotoxins, including alkylating agents, an oxidative agent, a ribonucleotide reductase inhibitor, a UV mimetic agent, an agent that causes double strand breaks, a topoisomerase I inhibitor, and ionizing radiation, all of which triggered a detectable and reproducible level of GFP production by the HUG1P -GFP strain. Furthermore, GFP was not induced by general cell stresses including starvation, heat shock, and acidic pH. These results suggest this system will be a valuable supplement to traditional genotoxicity assays. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Mutation Research - Genetic Toxicology and Environmental Mutagenesis Elsevier

The utilization of a Saccharomyces cerevisiae HUG1P -GFP promoter–reporter construct for the selective detection of DNA damage

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Publisher
Elsevier
Copyright
Copyright © 2007 Elsevier B.V.
ISSN
1383-5718
eISSN
1879-3592
D.O.I.
10.1016/j.mrgentox.2007.05.002
Publisher site
See Article on Publisher Site

Abstract

In this study, we report the creation and characterization of a yeast-based promoter–reporter construct for the detection of genotoxic compounds within a cell's local environment. We have synthesized a fusion containing the HUG1 promoter and GFP and incorporated this cassette into the yeast genome creating a stable, sensitive genotoxicity indicator. To quantify biosensor performance, HUG1P -GFP cells were exposed to multiple doses of a wide variety of genotoxins, including alkylating agents, an oxidative agent, a ribonucleotide reductase inhibitor, a UV mimetic agent, an agent that causes double strand breaks, a topoisomerase I inhibitor, and ionizing radiation, all of which triggered a detectable and reproducible level of GFP production by the HUG1P -GFP strain. Furthermore, GFP was not induced by general cell stresses including starvation, heat shock, and acidic pH. These results suggest this system will be a valuable supplement to traditional genotoxicity assays.

Journal

Mutation Research - Genetic Toxicology and Environmental MutagenesisElsevier

Published: Sep 1, 2007

References

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