Visualization and evaluation of the promoter activities of genes for stress-inducible proteins in response to environmental pollutants

Tomoko Ohashi, Tetsuya Satoh, Minoru Kuwata, Taira Fujimura, Shigeru Taketani

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    Abstract

    We examined the systematic assay of the reporter gene for the assessment of heavy metals and organic chemical pollutants using the reporter plasmids carrying stress-responsive elements fused to the green fluorescence protein (GFP) gene as follows: metallothionein (MTIIA), heme oxigenase-1 (HO-1), quinone reductase (ARE), and c-fos genes. The treatment of COS7 cells in which the c-fos gene promoter-, ARE-, or HO-1 enhancer-fused GFP with a low concentration of NaAsO2 was introduced led to the detection of the fluorescent cells, and an agrichemical paraquat enhanced the fluorescence of ARE or HO-1 enhancer-transfected cells. The cells in which the plasmid carrying the MT-IIA gene promoter (the -765 bp upstream from the transcription initiation site) was introduced highly expressed GFP on treatment with CdCl2, ZnSO 4, or CuCl2. The plasmid carrying seven metal-responsive elements of the MT-IIA gene increased the response of the fluorescence intensity to these heavy metals. These results indicated that the use of the gene promoters and enhancers of the stress-responsive genesfused to GFP contributes to the visualization of pollutantresponsive mammalian cells and can be applied to biomonitoring of environmental pollution.

    Original languageEnglish
    Pages (from-to)757-764
    Number of pages8
    JournalYakugaku Zasshi
    Volume127
    Issue number4
    DOIs
    Publication statusPublished - Apr 1 2007

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    All Science Journal Classification (ASJC) codes

    • Pharmacology
    • Pharmaceutical Science

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