2 A PROCESS FOR SCREENING POTENTIAL ENDOCRINE DISRUPTING CHEMICALS USING A YEAST TWO-HYBRID SYSTEM AND TRANSFORMED YEAST STRAINS THEREFOR

摘要

PURPOSE: A method for selecting putative endocrine disrupting chemicals(EDCs) using yeast two-hybrid system and transformed yeast used in the same method are provided, which method has similar sensitivity to the prior method using mammal cells, and the endocrine disrupting chemicals(EDCs) can be easily selected by the method. CONSTITUTION: The method for selecting putative endocrine disrupting chemicals(EDCs) using yeast two-hybrid system comprises the steps of: culturing transformed yeast having a first expression vector containing a first protein gene fused by a DNA-binding domain and a sex hormone receiptor or a ligand-binding domain of the sex hormone receiptor, a second expression vector expressing a second protein gene fused by a coactivator of the sex hormone receiptor and a transcription activating domain, and a third expression vector expressing a reporter gene operably linked to a recognition site of the DNA-binding domain together with putative endocrine disrupting chemical; and analyzing the transcription activity of the reporter gene to determine whether the putative endocrine disrupting chemical is antagonist or agonist of the reporter gene, wherein the sex hormone receiptor gene is androgen receiptor gene; the reporter gene is selected from -galactosidase, HIS3, Leu2 or URA3; the first vector is pLexA-ARhLBD; the second vector is pB42-ASC1; and the third vector is p8op-lacZ.