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Chromosome Architecture pp 31–41Cite as

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

Part of the Methods in Molecular Biology book series (MIMB,volume 2476)

Abstract

MukBEF, a structural maintenance of chromosome (SMC) complex, is an important molecular machine for chromosome organization and segregation in Escherichia 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.

Key words

  • Chromosome organization
  • MukBEF
  • E. coli
  • Fluorescence microscopy
  • FRAP

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Acknowledgments

M.L. acknowledges support from the Engineering and Physical Science Research Council (EPSRC EP/T002166/1, EP/N031431/1), the Biotechnology and Biological Sciences Research Council (BB/P000746/1, BB/R001235/1), and the Royal Society (IEC\NSFC\191406).

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Authors and Affiliations

  1. National Centre for Biological Sciences, GKVK Campus, Bengaluru, Karnataka, India

    Anjana Badrinarayanan

  2. Departments of Physics and Biology, University of York, York, UK

    Mark C. Leake

Authors
  1. Anjana Badrinarayanan
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  2. Mark C. Leake
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Corresponding author

Correspondence to Anjana Badrinarayanan .

Editor information

Editors and Affiliations

  1. Departments of Physics and Biology, University of York, York, UK

    Prof. Mark C. Leake

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Badrinarayanan, A., Leake, M.C. (2022). Fluorescence Recovery After Photobleaching (FRAP) to Study Dynamics of the Structural Maintenance of Chromosome (SMC) Complex in Live Escherichia coli Bacteria. In: Leake, M.C. (eds) Chromosome Architecture. Methods in Molecular Biology, vol 2476. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2221-6_4

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  • DOI: https://doi.org/10.1007/978-1-0716-2221-6_4

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  • Publisher Name: Humana, New York, NY

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