Abstract: AimTo establish a sensitive and efficient reporter gene-based screening model for finding agonists of estrogen receptor β subtype. MethodsA recombinant vector pTAL-ERE-SEAP was constructed by inserting a synthetic sequence composed of five estrogen responsive elements in front of promoter of pTAL-SEAP vector. pTAL-ERE-SEAP was then transfected into human embryonic kidney (HEK293) cells. G418 (200 μg·mL^-1) was added to select positive clones that can be induced by E₂ to express reporter gene SEAP. The speciality was tested by several ligands of relative nuclear receptors of the same family. The stability of the model, the time-effect relationship, the dose-response relationship, and the immunocytochemistry staining of ERβ expression after transfection were observed. 2 622 compounds were screened by using this model. ResultsStably transfected clones were obtained. The expression levels of reporter gene SEAP of positive clones was induced by E₂ in a dose-response and time-effect relationship manners. The Z′ factor value was 0.7. The expression levels of dexamethasone and other ligands were low. The result of immunocytochemistry staining showed the expression of ERβ. E₂ had no proliferating effects on stably transfected clones. ConclusionStably transfected positive clones transfected with recombinant vector pTAL-ERE-SEAP were obtained. The positive clones may be used to screen for agonists of estrogen receptor β subtype by measurement of luminescent value of expressed SEAP in wells of microlitre plate.