title: In Vitro Assembly of Nucleosomes for Binding/Remodeling Assays.
abstract: In order to study the functions and activities of chromatin remodeling enzymes in vitro, it is necessary to be able to reconstitute nucleosomes on DNA templates. In this chapter, we describe procedures for purification of histones from E. coli, formation of octamers, and reconstitution of nucleosomes that can be further modified by chromatin modifiers. In addition, we describe methods to purify nucleosomes from human cells. Finally, we also describe assays to test binding and remodeling activities of chromatin remodelers.
authors.affiliation: Department of Cell and Molecular Biology, University of Rhode Island, 287 CBLS, 120 Flagg Road, Kingston, RI, 02881, USA.
authors.forename: Arnob
authors.initials: A
authors.lastname: Dutta
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO, 64110, USA. jlw@stowers.org.
authors.forename: Jerry L
authors.initials: JL
authors.lastname: Workman
mesh_terms: D001681:Biological Assay / Q000379:methods*; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly / Q000235:genetics / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism*
chemical_list: D002843:Chromatin; D006657:Histones; D009707:Nucleosomes
keywords: Chromatin modifying enzyme; Chromatin remodeler; Histone; Nucleosome