| 细菌中优化冷激启动子的高通量筛选方法的建立 |
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| 引用本文： | 易永红,彭惠,高毅,孙敏,戴君.细菌中优化冷激启动子的高通量筛选方法的建立[J].工业微生物,2010,40(5):37-40. |
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| 作者姓名： | 易永红 彭惠 高毅 孙敏 戴君 |
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| 作者单位： | 安徽大学生命科学学院,安徽省生态工程与生物技术重点实验室,合肥,230039 |
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| 基金项目： | 国家自然科学基金，安徽高校省级自然科学研究项目，安徽大学博硕士队伍建设计划 |
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| 摘 要： | 为了优化冷激启动子,分别利用卡那霉素抗性基因kan和荧光蛋白(EGFP)基因为报告基因建立了两种高通量筛选方法。抗性筛选方法是通过观察菌落形态变化判断启动子强弱,适用于筛选强启动子。荧光筛选方法中,对转化子进行平板诱导,使用荧光显微镜拍摄菌落荧光,Matlab软件计算每一个菌落的亮度,实现高通量筛选,操作简单,精确度较好。
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| 关 键 词： | 高通量筛选 冷激启动子 抗性基因 荧光蛋白 |
Development of high-throughput screening for cold-shock promoters in bacteria |
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| YI Yong-hong,PENG Hui,GAO Yi,SUN Min,DAI Jun.Development of high-throughput screening for cold-shock promoters in bacteria[J].Industrial Microbiology,2010,40(5):37-40. |
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| Authors: | YI Yong-hong PENG Hui GAO Yi SUN Min DAI Jun |
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| Institution: | (Key Laboratory of Ecological Engineering and Bioteehnology of Anhui Province, School of Life Science, Anhui University, Hefei, 230039, China) |
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| Abstract: | Two different high-throughput screening for optimizing cold-shock promoters were established by using kanamycin （kan） resistance gene and enhanced green fluorescence protein （EGFP） gene as reporter, respectively. In resistance screening, the promoter strength was judged by observing the variations of colony morphology. This should, in principle, give us only strong promoters. In fluorescence screening, the fluorescence of transformant colonies was induced on plate and, subsequently, recorded under the fluorescence microscope. By applying Matlab, the brightness value of each colony was obtained. The results showed that the method was simple, rapid and valid. |
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| Keywords: | high-throughput screening cold-shock promoter resistant gene fluorescence protein |
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