ストレス蛋白質遺伝子のプロモーター活性の可視化と評価

概要

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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 NaAsO_2 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 CdCl_2, ZnSO_4, or CuCl_2. 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.
社団法人日本薬学会の論文
2007-04-01