关键词: 雌激素类化合物, EGFP, 表达载体, 瞬时转染

Abstract: The estrogenic endocrine disrupting compounds whether from environment and food chains or from chemicals illegally used in animal production pose a serious threat to the safety of animal derived products. Because of the huge diversities of these chemical structures, it is necessary to establish high-throughput screening and detecting methods to screen them. In this study, 4 recombination expression vectors with enhanced GFP as reporter gene were constructed. These constructs, namely, pERE-SV-EGFP, pERE-TA-EGFP, pERE-CMV-EGFP, pERE-TK-EGFP, only differ in promoters. Then the four expression vectors were transiently transfected into MCF-7 cells with liposome transfection reagent Tfx-20. The constitutive expressions of EGFP in transient transfected cells were related to the activity of the promoters in the expression vectors. And the fluorescence intensity treated by 17- βestradiol (10-9 mol·L-1) was two-fold higher than that treated by DMSO (control) in cells transiently transfected with pERE-SV-EGFP, pERE-TA-EGFP and pERE-TK-EGFP. These results indicated that the expression of reporter gene in ERE-SV-EGFP, pERE-TA-EGFP and pERE-TK-EGFP was estrogenic chemical dependent, and could be used for stable transfection to establish stable cell lines for estogenics screen.

Key words: Estrogenic chemicals, EGFP, Expression vector, Transient transfection