Yeast reporter system for rapid determination of estrogenic activity.
An in vitro test system for the determination of estrogens, xeno- and phytoestrogens, based on the activation of human estrogen receptor-alpha, has been examined for ability in monitoring environmental estrogens. The system consists of an expression plasmid for the human estrogen receptor-alpha and a reporter plasmid containing the lacZ gene under the control of the vitellogenin hormone response element. These plasmids have been transformed into S. cerevisae. Cultivation of yeast in the presence of estrogenic substances leads to activation of the estrogen receptor and induces the expression of the reporter lacZ. beta-Galactosidase activity of the translated gene lacZ is a measure of the estrogenic activity of a compound. First, the selectivity of the system was compared to data available in the literature. Then the sensitivity of the system was checked. The detection limit is 0.1 ng 17-beta estradiol or an equivalent activity per liter, if a sample can be concentrated 1000-fold. The system has been further characterized by selected compounds with known and unknown estrogenic activity.
Jungbauer
Alois
A
Institute for Applied Microbiology, University of Agricultural Sciences, Muthgasse 18 A-1190, Vienna, Austria. jungbaue@hp01.boku.ac.at
Beck
Verena
V
D000595:Amino Acid Sequence; D047628:Estrogen Receptor alpha; D004967:Estrogens / Q000494:pharmacology*; D017930:Genes, Reporter; D006801:Humans; D007763:Lac Operon; D010957:Plasmids; D011960:Receptors, Estrogen / Q000235:genetics; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012680:Sensitivity and Specificity
D047628:Estrogen Receptor alpha; D004967:Estrogens; D011960:Receptors, Estrogen