生物荧光方法快速测定环境水体中的甾体类激素  

A Rapid Sensitive Biosensor System for Rapid Determination of Flourescence Steroid

作　　者：于源华[1] 王静华[1] 宋禹[1] 杨佳新[1] 张昊[1] 刘红[2] 

机构地区：[1]长春理工大学,长春130022 [2]吉林大学口腔医院,长春130021

出　　处：《分析化学》2010年第7期1048-1051,共4页Chinese Journal of Analytical Chemistry

摘　　要：建立了环境中甾体类激素的生物荧光测定方法。利用睾丸酮丛毛单胞菌的生物酶基因与甾环结构特异的分子识别特性,结合绿色荧光蛋白(GFP)的荧光标记技术,通过基因同源重组技术建立了特异性生物荧光检测系统,实现对环境中甾环类化合物的灵敏监测。制备绿色荧光突变菌及总蛋白含量为1.0g/L的非细胞体系工作液,在室温条件下对不同浓度雌二醇标准品进行检测,10min即可检测出fg级的甾体类激素,最佳检测时间为30min,方法的线性范围1.0～266ng/L,灵敏度高于高效液相色谱方法。本方法可应用于环境的实际检验工作。A bio-fluorescence method for the determination of steroid hormone in environment substance was established.Enzyme can specifically recognize steroid ring structure of hormone-steroid.Green fluorescent protein （GFP） has a report-phenomeon.On the base of these two features,a bio-induced fluorescence detection system was established through the genetic homologous recombination techniques,the research achieved the specificity and sensitivity of the monitoring of steroids.After preparing GFP mutant strains and cell-free system working solution containing 1.0 g/L protein,the estradiol standard steroid in different concentrations was detected at room temperature.The results show that the fg level of steroid hormone can be detected within 10 min at room temperature.The detection time can maintain stable in 30 min.The linear range is 0.001-0.266 fg/mL.This method is more sensitive than HPLC with six orders of magnitude.The method is sensitive,simple,accurate,specificity,and can be applied to the inspection of steroid in environment substance.

关 键 词：甾体类激素 生物荧光 基因同源重组 分子识别 

分 类 号：X832[环境科学与工程—环境工程]