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Fig. 3 | BMC Evolutionary Biology

Fig. 3

From: Conservation of estrogen receptor function in invertebrate reproduction

Fig. 3

Chemical Complementation is a binding assay. This assay couples yeast survival to the presence of a small molecule ligand (red). The yeast contains the GAL4 response element that controls the expression of HIS3 biosynthesis genes. The rotifer putative ER LBD is fused to a GAL4 DNA binding domain (GBD). This fusion protein binds to the GAL4 response element. Ligand binding by the ER LBD leads to recruitment of the SRC-1 coactivator, which is fused to the GAL4 activation domain (GAD). Transcription of the histidine biosynthesis genes, upon ligand binding, allows the yeast to survive in media lacking histidine [37,38,39,40]. The results show that estradiol and the synthetic fluorescent ligands bind to the rotifer the ER-like LBD in vivo. The negative control (with no added ligand) exhibited minimal growth. The natural ligand, estradiol, increases growth on the same order as the synthetic ligands. Neither estradiol, nor the synthetic fluorescent compounds have any effect on yeast growth in the absence of Gal4-ER [35]

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