Using Fluorescence Recovery After Photobleaching (FRAP) to Study Dynamics of the Structural Maintenance of Chromosome (SMC) Complex In Vivo.
The SMC complex, MukBEF, is important for chromosome organization and segregation in Escherichia coli. Fluorescently tagged MukBEF forms distinct spots (or "foci") in the cell, where it is thought to carry out most of its chromosome associated activities. This chapter outlines 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.
Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave, Cambridge, MA, 02139, USA. anjana1@mit.edu.
Anjana
A
Badrinarayanan
Biological Physical Sciences Institute (BPSI), University of York, Heslington, York, YO10 5DD, UK.
Mark C
MC
Leake
D001426:Bacterial Proteins / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D002876:Chromosomes, Bacterial; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism*; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods*; D008164:Luminescent Proteins / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D001426:Bacterial Proteins; D002868:Chromosomal Proteins, Non-Histone; D029968:Escherichia coli Proteins; D008164:Luminescent Proteins; C067035:MukB protein, E coli; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria
Chromosome organization; E. coli; FRAP; Fluorescence microscopy; MukBEF