In vitro and in vivo methodologies for studying the Sigma 54-dependent transcription.
In vitro and in vivo methodologies for studying the Sigma 54-dependent transcription.
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用于研究 Sigma 54 依赖性转录的体外和体内方法。
DOI：
10.1007/978-1-4939-2392-2_4
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Buck M
中科院分区：
文献类型：
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作者：
Buck M
在这里，我们描述了在纯化系统中体外测定主要变体细菌σ因子σ 54（σ54）的途径和方法。我们包括完整的转录系统，σ54与其激活剂之间的结合相互作用，以及用于重塑封闭启动子复合物的同源激活剂的自组装和关键的ATP酶活性。我们还提出了用于研究生理过程、代谢状态、全局信号网络和细胞结构对σ54依赖性基因表达控制的影响的体内方法。
Here we describe approaches and methods to assaying in vitro the major variant bacterial sigma factor, Sigma 54 (σ54), in a purified system. We include the complete transcription system, binding interactions between σ54 and its activators, as well as the self-assembly and the critical ATPase activity of the cognate activators which serve to remodel the closed promoter complexes. We also present in vivo methodologies that are used to study the impact of physiological processes, metabolic states, global signalling networks, and cellular architecture on the control of σ54-dependent gene expression.