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.
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Acknowledgments
We thank current and former members of the Workman laboratory for help with developing these protocols and many useful discussions. We acknowledge Malini Natarajan for critical reading of the manuscript. This work was supported by National Institutes of Health grant R01GM047867 to J.L.W. and funding from the Stowers Institute for Medical Research.
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Dutta, A., Workman, J.L. (2017). In Vitro Assembly of Nucleosomes for Binding/Remodeling Assays. In: Guillemette, B., Gaudreau, L. (eds) Histones. Methods in Molecular Biology, vol 1528. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6630-1_1
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DOI: https://doi.org/10.1007/978-1-4939-6630-1_1
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Online ISBN: 978-1-4939-6630-1
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