荧光标记的单链结合蛋白在体内定位实验中的应用。

Use of fluorescently tagged SSB proteins in in vivo localization experiments.

作者信息

Reyes-Lamothe Rodrigo

机构信息

Department of Biochemistry, University of Oxford, Oxford, UK.

出版信息

Methods Mol Biol. 2012;922:245-53. doi: 10.1007/978-1-62703-032-8_19.

DOI:10.1007/978-1-62703-032-8_19

PMID:22976192

Abstract

The time and place of DNA replication in cells provides invaluable insight into the cell cycle and DNA metabolism. An effective means of obtaining this information is through fluorescence microscopy. The abundance of Single-Strand Binding protein, SSB, at the replication fork makes it a good reporter of DNA replication. In this chapter I describe how to observe replication of the Escherichia coli chromosome in a strain that synthesizes a fluorescent derivative of SSB. This methodology provides information about the position and dynamics of DNA replication through epifluorescence.

摘要

细胞中DNA复制的时间和位置为深入了解细胞周期和DNA代谢提供了极有价值的信息。获取这些信息的一种有效方法是通过荧光显微镜观察。单链结合蛋白（SSB）在复制叉处的大量存在使其成为DNA复制的良好报告分子。在本章中，我将描述如何在一个合成了SSB荧光衍生物的大肠杆菌菌株中观察其染色体的复制。这种方法通过落射荧光提供了有关DNA复制位置和动态的信息。