Environmental pollution of estrogenic compounds, which are known endocrine disrupting chemicals, creates the need for an easy and sensitive assay for their detection. We developed a protein complementation assay in which two split fragments of yeast cytosine deaminase (yCD) were fused to the estrogen receptor a ligand binding domain (ER-LBD) and the conformational peptide, a�/I, respectively. The conformational peptide recognizes the 17-� estradiol bound form of ER-LBD, and thus in the presence of 17-� estradiol, yCD is reconstituted. The recovered yCD activity was monitored through a whole-cell life and death assay using the prodrug, 5-fluorocytosine. A dose-dependent growth inhibition was observed with a picomolar detection limit for 17-� estradiol. This simple in vivo assay can be applied towards the detection of other estrogenic compounds as well as for use in estrogen receptor drug screening.

Development of a highly sensitive split protein based whole-cell bioassay to detect estrogenic compounds

247th National Meeting (2014)

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