Dithiocarbamate fungicide thiram detection: comparison of bioluminescent and fluorescent whole-cell bioassays based on hsp22 stress promoter induction.
Detection of toxic substances interfering with endocrine system is one of the major preoccupations of the European community. A whole-cell bioassay for pollution detection based on stress induction has been designed. Well characterized toxicants, cadmium chloride and thiram (a dithiocarbamate fungicide), were used to optimize the detection conditions such as time-course conditions, cell line and reporter gene to be used. HeLa cells containing the firefly luciferase (luc) reporter gene under the control of the Drosophila melanogaster hsp22 promoter were compared to liver cells (HepG2) containing the same stress gene promoter fused either to the luc or the EGFP (Enhanced-Green Fluorescent Protein) gene. The sensitivity of the obtained bioassay was found to be enhanced by the concomitant use of liver cells and EGFP reporter gene. The detection limits of the toxicants were then lowered from 1 to 0.1 microM and from 1 to 0.01 microM for CdCl(2) and thiram, respectively.
Mandon
C A
CA
Laboratoire de Génie Enzymatique et Biomoléculaire, UMR 5013 CNRS, Université Claude Bernard Lyon 1, Villeurbanne, France. mandon@univ-lyon1.fr
Diaz-Latoud
C
C
Arrigo
A-P
AP
Blum
L J
LJ
D001681:Biological Assay / Q000379:methods*; D019256:Cadmium Chloride / Q000032:analysis* / Q000633:toxicity; D029721:Drosophila Proteins / Q000235:genetics; D004785:Environmental Pollutants / Q000032:analysis* / Q000633:toxicity; D005659:Fungicides, Industrial / Q000032:analysis* / Q000633:toxicity; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells / Q000187:drug effects / Q000201:enzymology; D006360:Heat-Shock Proteins / Q000235:genetics; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism*; D013893:Thiram / Q000032:analysis* / Q000633:toxicity
D029721:Drosophila Proteins; D004785:Environmental Pollutants; D005659:Fungicides, Industrial; D006360:Heat-Shock Proteins; C490752:Hsp22 protein, Drosophila; D013893:Thiram; D049452:Green Fluorescent Proteins; D049409:Luciferases, Firefly; D019256:Cadmium Chloride