From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression
From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression
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DOI：
10.1016/j.jmb.2012.01.001
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发表时间：
2012-03-02
影响因子：
5.6
通讯作者：
Moeglich, Andreas
中科院分区：
文献类型：
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作者：
Ohlendorf, Robert;Vidavski, Roee R.;Moeglich, Andreas
信号感光体响应光介导多种有机体适应。作为光门控蛋白开关，信号传导感受器为光遗传学提供了基础，该术语指的是通过光控制有机体生理和行为。我们建立了新型的光遗传学工具PDUSK和PDAWN，它们采用蓝光光感受器在大肠杆菌中赋予光抑制或光诱导的基因表达，并在照明时诱导高达460倍。这些系统的关键特征是背景活动较低，动态范围高，在20 mu尺度上的空间控制，远离外源性因素以及易用性。在光遗传实验中，PDUSK和PDAWN可用于特异性扰动信号网络的单个节点并询问其作用。在制备量表上，Pdawn可以通过灯光诱导重组蛋白的产生，因此代表了传统诱导系统的成本效益且易于自动化的替代品。 （c）2012 Elsevier Ltd.保留所有权利。
Signaling photoreceptors mediate diverse organismal adaptations in response to light. As light-gated protein switches, signaling photoreceptors provide the basis for optogenetics, a term that refers to the control of organismal physiology and behavior by light. We establish as novel optogenetic tools the plasmids pDusk and pDawn, which employ blue-light photoreceptors to confer light-repressed or light-induced gene expression in Escherichia coli with up to 460-fold induction upon illumination. Key features of these systems are low background activity, high dynamic range, spatial control on the 20-mu m scale, independence from exogenous factors, and ease of use. In optogenetic experiments, pDusk and pDawn can be used to specifically perturb individual nodes of signaling networks and interrogate their role. On the preparative scale, pDawn can induce by light the production of recombinant proteins and thus represents a cost-effective and readily automated alternative to conventional induction systems. (C) 2012 Elsevier Ltd. All rights reserved.