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

Anjana Badrinarayanan, Mark C Leake

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)31-41
Number of pages11
JournalMethods in Molecular Biology
Volume2476
DOIs
Publication statusPublished - 31 May 2022

Bibliographical note

© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature. This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details

Keywords

  • Chromosomal Proteins, Non-Histone/genetics
  • Chromosomes
  • Escherichia coli/chemistry
  • Escherichia coli Proteins/chemistry
  • Fluorescence Recovery After Photobleaching
  • Repressor Proteins/genetics

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