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PMID: 10648917 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Application of yeast cells transformed with GFP expression constructs containing the RAD54 or RNR2 promoter as a test for the genotoxic potential of chemical substances.

Mutation research ·Vol. 464 ·No. 2 ·2000-01-24 ·Pages 297-308

Afanassiev V, Sefton M, Anantachaiyong T, Barker G, Walmsley R, Wölfl S

Abstract

Yeast strains transformed with high copy number plasmids carrying the gene encoding a green fluorescent protein optimised for yeast (yEGFP3) under the control of the RAD54 or RNR2 promoter were used to investigate the activity of potentially DNA-damaging substances. The assays were performed on 96-well microtitre plates in the presence of different concentrations of the test substances. The synthesis of GFP protein was measured through the fluorescence signal and cell growth was monitored by absorption. Here, we demonstrate that this system can be used as a biosensor to assess the genotoxic potential of drugs and other chemical substances. The use of microtitre plates will enable full automation of the system and allows the inclusion of internal reference standards in each assay.

MeSH Terms
Biosensing Techniques Blotting, Western DNA Helicases DNA Repair Enzymes Dose-Response Relationship, Drug Enzyme Inhibitors/toxicity Evaluation Studies as Topic Fluorescence Fungal Proteins/genetics Gene Expression/drug effects Genes, Reporter/drug effects Green Fluorescent Proteins Luminescent Proteins/genetics Methyl Methanesulfonate/toxicity Mutagenicity Tests/methods Mutagens/toxicity Predictive Value of Tests Promoter Regions, Genetic/genetics Ribonucleotide Reductases/genetics Saccharomyces cerevisiae/drug effects,genetics Saccharomyces cerevisiae Proteins Sensitivity and Specificity Transfection
Chemicals
Enzyme Inhibitors Fungal Proteins Luminescent Proteins Mutagens Saccharomyces cerevisiae Proteins Green Fluorescent Proteins Methyl Methanesulfonate Ribonucleotide Reductases ribonucleotide reductase R2 subunit RAD54 protein, S cerevisiae DNA Helicases DNA Repair Enzymes
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Afanassiev V
Hans-Knöll-Institut für Naturstoff-Forschung, Abteilung Zell- und Molekularbiologie, Beutenbergstr. 11, D-07745, Jena, Germany.
Sefton M
Anantachaiyong T
Barker G
Walmsley R
Wölfl S
Article Info
Journal
Mutation research
Abbr.
Mutat Res
ISSN
0027-5107
Published
2000-01-24
Pages
297-308
Language
English
Region
Netherlands
NLM ID
0400763
Subset
IM
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