|本期目录/Table of Contents|

[1]刘玉倩,魏婷,张超,等. 超敏感酵母HUG1-yEGFP生物传感器遗传毒性化合物检测方法优化[J].应用与环境生物学报,2014,20(05):919-924.[doi:10.3724/SP.J.1145.2014.01041]
 LIU Yuqian,WEI Ting,ZHANG Chao,et al. An optimized genotoxin detection method based on super-sensitive yeast HUG1-yEGFP biosensor[J].Chinese Journal of Applied & Environmental Biology,2014,20(05):919-924.[doi:10.3724/SP.J.1145.2014.01041]





 An optimized genotoxin detection method based on super-sensitive yeast HUG1-yEGFP biosensor
 刘玉倩 魏婷 张超 张晓华 袁丽 戴和平
1中国科学院水生生物研究所淡水生态和生物技术国家重点实验室 武汉 430072
2中国科学院大学 北京 100049
 LIU Yuqian WEI Ting ZHANG Chao ZHANG Xiaohua YUAN Li DAI Heping
1Institute of Hydrobiology, State Key Laboratory of Freshwater Ecology and Biotechnology, Chinese Academy of Sciences, Wuhan 430072, China
2University of Chinese Academy of Sciences, Beijing 100049, China
 biosensor yeast genotoxin fluorescence induction
Q3-3 : X82
基于酵母DNA损伤响应机制,本实验室构建了超敏感型酵母HUG1-yEGFP生物传感器,检测方法主要依靠流式细胞仪. 为了简化检测方法,实现高通量检测,本研究通过摸索和测试各种条件,建立了优化的、更适合于96孔板的荧光酶标仪的检测方法. 结果显示:F1培养基可以作为一种本底荧光较低的培养基用于荧光酶标仪的检测,使用96孔透明底黑色细胞培养板的检测菌液体积为100 μL,初始菌液浓度OD600 nm值为0.05. 在此优化检测条件下,超敏感酵母HUG1-yEGFP生物传感器与典型的遗传毒性化合物甲磺酸甲酯(MMS)可以建立良好的剂效和时效关系,说明该生物传感器已具备了实际应用的实验基础.
A super-sensitive yeast HUG1-yEGFP biosensor based on DNA damage response has been constructed in our laboratory. The biosensor was detected mainly by flow cytometer. This study aimed to simplify the detection by constructing a new detection assay with high throughput and optimize the detection conditions to adapt to 96-well microtiter plate. The super-sensitive yeast cells carrying genotoxic detecting biosensors were treated with MMS. The yEGFP-fluorescent intensity was measured by multifunctional fluorescent microplate reader. The result found F1 as the best yeast culture medium with low background of fluorescence for microplate fluorescent detection. For a high throughput genotoxin detection using 96-well microtiter plates with black walls and transparent bottoms, the best culture volume of yeast was 100 μL, and initial optical density at 600nm was 0.05. The dose-effect and time-effect between super-sensitive yeast biosensor and MMS under the optimized detection conditions showed that this biosensor is promising for practical application.


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 国家自然科学基金重点项目(21037004)与淡水生态和生物技术国家重点实验室项目(2012FBZ10)资助 Supported by the National Natural Science Foundation of China (21037004) and the State Key Laboratory of Freshwater Ecology and Biotechnology (2012FBZ10)
更新日期/Last Update: 2014-10-30