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Improvement of the Biosensor for Detection of Endocrine Disruptors by Combination of Human Estrogen Receptorα and Co-Activator  

Lee, Haeng-Seog (안양대학교 해양생명공학과)
Publication Information
Journal of Korean Society of Water and Wastewater / v.20, no.6, 2006 , pp. 893-904 More about this Journal
Abstract
To improve sensitivity of biosensor as yeast two-hybrid detection system for estrogenic activity of suspected chemicals, we tested effects of several combinations of the bait and fish components in the two-hybrid system on Saccharomyces cerevisiae inducted a chromosome-integrated lacZ reporter gene that was under the control of CYC1 promoter and the upstream Gal4p-binding element $UAS_{GAL}$. The bait components that were fused with the Gal4p DNA binding domain are full-length human estrogen receptor ${\alpha}$ and its ligand-binding domain. The fish components that were fused with the Gal4p transcriptional activation domain were nuclear receptor-binding domains of co-activators SRC1 and TIF2. We found that the combination of the full-length human estrogen receptor ${\alpha}$ with the nuclear receptor-binding domain of co-activator SRC1 was most effective for the estrogen-dependent induction of reporter activity among the two-hybrid systems so far reported. The relative strength of transcriptional activation by representative natural and xenobiotic chemicals was well correlated with their estrogenic potency that had been reported with other assay systems.
Keywords
endocrine disruptors; biosensor; yeast two-hybrid system; $hER{\alpha}$; co-activator;
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