关键词: 环境雌激素效应, 重组基因酵母, 受体作用

Abstract: The environmental hormones with accumulation and potential risk were concerned recently because of the inherent toxicity. The testing methods of the environmental hormones were studied by several research. Chemical methods were the major testing methods to determine the known environmental hormones during the past few years. In this paper we established and optimized a biological testing method based on the receptor effects theory. The experimental parameters of environmental hormone effects testing based on the receptor effects theory were optimized, including recovery and proliferation of recombinant gene yeast, exposure time and temperature of estrogenic compounds, the testing method of β- galactosidase, the optimum testing temperature, colorimetric time and colorimetric wavelength, statistical method by ORIGIN, and quantitative method, to analyze the estrogenic effect, in which 17β-estradiol was used as the standard reference materials. The results indicated that the method was more efficiently and accurately to determine the actual samples when recovery and proliferation of recombinant gene yeast was cultured in SD/-Trp/-Leu auxotrophic medium and shaked in 30℃ after 66 h when the yeast initiated the logarithmic growth. We could mix sample and yeast mixture in 20℃-35℃ water bath for 3-6 hours to improve the sensitivity.The β- Galactosidase should be released by organic solvent treatment in 35℃ water batch for 60 min, then 420 nm colorimetric analysis after 2.5 hours. The dose-effect curves of the environmental hormones fitted by ORIGIN were best when the concentration were between 0.025 nmol·L^-1 to 0.25 nmol·L^-1. In addition, we suggested that the dilution sample should be added in actual tests to increase the accuracy of the methods.

Key words: Estrogenic effect, gene recombined yeast, receptor effects