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目的建立环境雄激素内分泌干扰物酵母评价体系。方法以载体双表达的方式构建该重组基因酵母细胞。在表达载体中,用3-磷酸甘油醛脱氢酶(GPD)启动子驱动雄激素受体基因(AR)的表达,并使其与V5抗原表位相融合;在报告载体中,用雄激素效应元件(ERE)调控的Lac Z作为报告基因。将两者转化于酵母细胞(W303-1A)中,构建成雄激素调控的重组Lac Z基因酵母细胞。用不同浓度的雄激素[去氢表雄酮(DHT)和丙酸睾酮(TP)]和雌激素(雌二醇、雌三醇、雌酮、二乙基已烯雌酚和乙炔基雌二醇),对重组基因酵母细胞分别进行敏感度和特异度研究。结果去氢表雄酮和丙酸睾酮与该重组基因酵母细胞有明显的剂量效应关系,说明其具有良好的特异度,而与雌二醇、雌三醇、雌酮、二乙基已烯雌酚和乙炔基雌二醇5种雌激素化合物均无明显的剂量效应关系,说明其具有较强的特异性。结论该重组基因酵母细胞可用于对环境雄激素类化合物的筛选。
Objective To establish an environmental androgen endocrine disruptor yeast evaluation system. Methods The recombinant yeast cells were constructed by double expression of vectors. In the expression vector, the expression of the androgen receptor gene (AR) is driven by the glyceraldehyde-3 -phosphate dehydrogenase (GPD) promoter and fused to the V5 epitope; in the reporter vector the androgen effect Element (ERE) regulates LacZ as a reporter. The two were transformed into yeast cells (W303-1A) to construct androgen-regulated recombinant LacZ yeast cells. Effects of different concentrations of androgen [dehydroepiandrosterone (DHT) and testosterone propionate (TP)] and estrogens (estradiol, estriol, estrone, diethylstilbestrol and ethynylestradiol Alcohol), respectively, for the study of the sensitivity and specificity of recombinant yeast cells. Results Dehydroepiandrosterone and testosterone propionate had obvious dose-effect relationship with the recombinant yeast cells, indicating that they had good specificity, but not with estradiol, estriol, estrone, diethylhexolene Phenol and ethynyl estradiol 5 estrogenic compounds have no significant dose-response relationship, indicating that it has strong specificity. Conclusion The recombinant yeast cells can be used for the screening of environmental androgenic compounds.