| Chapter title |
Polycomb Group Proteins
|
|---|---|
| Chapter number | 13 |
| Book title |
Polycomb Group Proteins
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-6380-5_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6378-2, 978-1-4939-6380-5
|
| Authors |
Cherubini, Alessandro, Zippo, Alessio, Alessandro Cherubini, Alessio Zippo |
| Editors |
Chiara Lanzuolo, Beatrice Bodega |
| Abstract |
The Polycomb group (PcG) proteins form regulatory complexes that modify the chromatin structure and silence their target genes. Recent works have found that the composition of Polycomb complexes is highly dynamic. Defining the different protein components of each complex is fundamental for better understanding their biological functions. Fluorescent resonance energy transfer (FRET) is a powerful tool to measure protein-protein interactions, in nanometer order and in their native cellular environment. Here we describe the preparation and execution of a typical FRET experiment using CFP-tagged protein as donor and YFP-tagged protein as acceptor. We further show that FRET can be used in a competition assay to measure binding affinities of different components of the same chromatin complex. |
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