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We developed a yeast two-hybrid assay using a human-type estrogen receptor (hERα) to screen test chemicals reporting weak estrogenic activity at high concentrations. As a result of exposure to several test chemicals, viable yeast cells showed a decrease in num- ber compared with a control. It was shown that this phenomenon was due to fungicidal effects of the test chemicals within the yeast cell or to inhibition of yeast cell proliferation. Therefore, in performing a yeast two- hybrid assay, it is necessary to monitor yeast cell pro- liferation by methods such as the measurement of the number of viable cells, and it is necessary to screen test chemicals within a concentration range where these chemicals show neither fungicidal effects towards the yeast cell nor inhibition of yeast cell proliferation.

Problems with Methods used to Screen Estrogenic Chemicals by Yeast Two-Hybrid Assays.