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

Victor Afanassiev, Mark Sefton, Thaddao Anantachaiyong, Gordon Barker, Richard Walmsley, Stefan Wölfl

    Research output: Contribution to journalArticlepeer-review

    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. Copyright (C) 2000 Elsevier Science B.V.
    Original languageEnglish
    Pages (from-to)297-308
    Number of pages11
    JournalMutation Research - Genetic Toxicology and Environmental Mutagenesis
    Volume464
    Issue number2
    DOIs
    Publication statusPublished - 24 Jan 2000

    Keywords

    • Chemical substances
    • DNA damage
    • Genotoxicity
    • GFP reporter
    • MMS
    • Yeast

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