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在体阿拉伯糖启动子的激活和后续活性的单分子动力学。

In vivo single-molecule kinetics of activation and subsequent activity of the arabinose promoter.

机构信息

Laboratory of Biosystem Dynamics, Computational Systems Biology Research Group, Department of Signal Processing, Tampere University of Technology, FI-33101 Tampere, Finland.

出版信息

Nucleic Acids Res. 2013 Jul;41(13):6544-52. doi: 10.1093/nar/gkt350. Epub 2013 May 3.

Abstract

Using a single-RNA detection technique in live Escherichia coli cells, we measure, for each cell, the waiting time for the production of the first RNA under the control of PBAD promoter after induction by arabinose, and subsequent intervals between transcription events. We find that the kinetics of the arabinose intake system affect mean and diversity in RNA numbers, long after induction. We observed the same effect on Plac/ara-1 promoter, which is inducible by arabinose or by IPTG. Importantly, the distribution of waiting times of Plac/ara-1 is indistinguishable from that of PBAD, if and only if induced by arabinose alone. Finally, RNA production under the control of PBAD is found to be a sub-Poissonian process. We conclude that inducer-dependent waiting times affect mean and cell-to-cell diversity in RNA numbers long after induction, suggesting that intake mechanisms have non-negligible effects on the phenotypic diversity of cell populations in natural, fluctuating environments.

摘要

利用一种单 RNA 检测技术在活大肠杆菌细胞中,我们测量了每个细胞在诱导阿拉伯糖后由 PBAD 启动子控制的第一个 RNA 的产生的等待时间,以及转录事件之间的后续间隔。我们发现,阿拉伯糖摄取系统的动力学在诱导后很长时间内影响 RNA 数量的平均值和多样性。我们在 Plac/ara-1 启动子上观察到了相同的效果,该启动子可以被阿拉伯糖或 IPTG 诱导。重要的是,如果仅由阿拉伯糖诱导,则 Plac/ara-1 的等待时间分布与 PBAD 的分布无法区分。最后,发现 PBAD 控制下的 RNA 产生是一个亚泊松过程。我们得出结论,诱导剂依赖性等待时间在诱导后很长时间内影响 RNA 数量的平均值和细胞间多样性,这表明摄取机制对自然波动环境中细胞群体的表型多样性有不可忽视的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b86/3711423/338febf7ca2d/gkt350f1p.jpg

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