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Methods Mol Biol. 2017;1528:1-17.

In Vitro Assembly of Nucleosomes for Binding/Remodeling Assays.

Author information

1
Department of Cell and Molecular Biology, University of Rhode Island, 287 CBLS, 120 Flagg Road, Kingston, RI, 02881, USA.
2
Stowers Institute for Medical Research, Kansas City, MO, 64110, USA. jlw@stowers.org.

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.

KEYWORDS:

Chromatin modifying enzyme; Chromatin remodeler; Histone; Nucleosome

PMID:
27854012
DOI:
10.1007/978-1-4939-6630-1_1
[Indexed for MEDLINE]

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