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Single-Stranded DNA Binding Proteins pp 245–253Cite as

Use of Fluorescently Tagged SSB Proteins in In Vivo Localization Experiments

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 922))

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.

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Acknowledgment

I am grateful to A. Badrinarayanan, C. Lesterlin, N. Paramio, M. Tolmasky, and D. Sherratt for valuable comments on the manuscript. This work was by supported by the Welcome Trust and a research fellowship of New College (Oxford).

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Authors and Affiliations

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

    Rodrigo Reyes-Lamothe

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  1. Rodrigo Reyes-Lamothe
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Correspondence to Rodrigo Reyes-Lamothe .

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Editors and Affiliations

  1. School of Medicine and Public Health, Department of Biomolecular Chemistry, University of Wisconsin, Henry Mall 420, Madison, 53706, USA

    James L. Keck

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Reyes-Lamothe, R. (2012). Use of Fluorescently Tagged SSB Proteins in In Vivo Localization Experiments. In: Keck, J. (eds) Single-Stranded DNA Binding Proteins. Methods in Molecular Biology, vol 922. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-032-8_19

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  • DOI: https://doi.org/10.1007/978-1-62703-032-8_19

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-031-1

  • Online ISBN: 978-1-62703-032-8

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