[Visualization and evaluation of the promoter activities of genes for stress-inducible proteins in response to environmental pollutants].
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 genes fused to GFP contributes to the visualization of pollutant-responsive mammalian cells and can be applied to biomonitoring of environmental pollution.
Ohashi
Tomoko
T
Hyogo Prefectural Institute of Technology, Kobe, Japan. tomoko@hyogo-kg.go.jp
Satoh
Tetsuya
T
Kuwata
Minoru
M
Fujimura
Taira
T
Taketani
Shigeru
S
D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004742:Enhancer Elements, Genetic; D004784:Environmental Monitoring / Q000379:methods; D004785:Environmental Pollutants; D050939:Gene Fusion; D017930:Genes, Reporter; D016762:Genes, fos / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D051547:Heme Oxygenase-1 / Q000235:genetics*; D008668:Metallothionein / Q000235:genetics*; D019216:Metals, Heavy; D051379:Mice; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011817:Rabbits; D051381:Rats
D004785:Environmental Pollutants; D019216:Metals, Heavy; D049452:Green Fluorescent Proteins; D008668:Metallothionein; D051547:Heme Oxygenase-1; D016660:NAD(P)H Dehydrogenase (Quinone)