The accumulating discoveries of new posttranslational modifications (PTMs) and the increasing relevance of histone variants within the frame of epigenetics demand the availability of methods for a rapid and efficient nucleosome reconstitution to analyze their structural and functional implications. Here we describe a method suitable for this purpose, starting from bacterially expressed histones, solubilized by acid and purified by reversed-phase high-performance liquid chromatography. This method allows the preparation of micrograms to milligram amounts of in vitro-assembled nucleosomes. Finally, we demonstrate the efficiency of this method for the structural analysis of nucleosomes in the analytical ultracentrifuge.
CITATION STYLE
Cheema, M. S., & Ausió, J. (2017). Analytical ultracentrifuge analysis of nucleosomes assembled from recombinant, acid-extracted, HPLC-purified histones. In Methods in Molecular Biology (Vol. 1528, pp. 75–95). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6630-1_6
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