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利用双荧光报告系统分析启动子激活中的随机性。

Analysis of stochasticity in promoter activation by using a dual-fluorescence reporter system.

作者信息

Ou Jianhong, Furusawa Chikara, Yomo Tetsuya, Shimizu Hiroshi

机构信息

Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, 565-0871, Japan.

出版信息

Biosystems. 2009 Sep;97(3):160-4. doi: 10.1016/j.biosystems.2009.06.002. Epub 2009 Jun 23.

Abstract

Stochastic dynamics of promoter activity in bacterial cells were studied by using a dual-fluorescence reporter system of protein expression. The dual-fluorescence reporter system enabled us to calculate the amplitude of intrinsic noise generated during transcription and translation. By fitting the experimental data to a simple stochastic model of protein expression, we could estimate parameters representing the stochastic transition between the active and inactive states of a promoter. Using the system, we analyzed the stochastic dynamics of promoter activation of genes in the lysine biosynthesis pathway in Escherichia coli. We found that the promoter of lysA has a significantly slower transition rate between active and inactive states than other promoters in the lysine biosynthesis pathway. The infrequent switching between active and inactive states can be a dominant source of noise in lysA expression. Analysis using the dual-fluorescence reporter system provided a better understanding of stochastic dynamics in promoter activation.

摘要

通过使用蛋白质表达的双荧光报告系统,研究了细菌细胞中启动子活性的随机动力学。双荧光报告系统使我们能够计算转录和翻译过程中产生的内在噪声的幅度。通过将实验数据拟合到一个简单的蛋白质表达随机模型,我们可以估计代表启动子活性和非活性状态之间随机转变的参数。使用该系统,我们分析了大肠杆菌赖氨酸生物合成途径中基因启动子激活的随机动力学。我们发现,与赖氨酸生物合成途径中的其他启动子相比,lysA的启动子在活性和非活性状态之间的转变速率明显较慢。活性和非活性状态之间的不频繁切换可能是lysA表达中噪声的主要来源。使用双荧光报告系统进行的分析有助于更好地理解启动子激活中的随机动力学。

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