Fluorescence Recovery After Photobleaching (FRAP) to Study Dynamics of the Structural Maintenance of Chromosome (SMC) Complex in Live Escherichia coli Bacteria.

Fluorescence Recovery After Photobleaching (FRAP) to Study Dynamics of the Structural Maintenance of Chromosome (SMC) Complex in Live Escherichia coli Bacteria.
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光漂白后的荧光恢复 (FRAP) 用于研究活大肠杆菌中染色体 (SMC) 复合物结构维持的动态。

DOI:
10.1007/978-1-0716-2221-6_4
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发表时间:
2022
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
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通讯作者:
Badrinarayanan A
Badrinarayanan A
中科院分区:
--
文献类型:
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作者:
Badrinarayanan A

文献摘要

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MukBEF 是一种染色体结构维护 (SMC) 复合体,是大肠杆菌中染色体组织和分离的重要分子机器。荧光标记的 MukBEF 在细胞中形成由分子组装体组成的独特斑点(或“焦点”),人们认为它在此处执行大部分与染色体相关的活动。在这里,我们概述了光漂白后荧光恢复 (FRAP) 技术作为研究荧光焦点中 YFP 标记的 MukB 特性的方法。该方法可以为 MukB 在 DNA 上的动态提供重要的见解,并可用于研究其体内生化特性。
MukBEF, a structural maintenance of chromosome (SMC) complex, is an important molecular machine for chromosome organization and segregation inEscherichia coli.Fluorescently tagged MukBEF forms distinct spots (or “foci”) composed of molecular assemblies in the cell, where it is thought to carry out most of its chromosome-associated activities. Here, we outline the technique of fluorescence recovery after photobleaching (FRAP) as a method to study the properties of YFP-tagged MukB in fluorescent foci. This method can provide important insight into the dynamics of MukB on DNA and be used to study its biochemical properties in vivo.