Production and Assay of Recombinant Multisubunit Chromatin Remodeling Complexes.

Methods Enzymol

Section of Structural Biology, Imperial College London, London, United Kingdom. Electronic address:

Published: April 2018

We have developed a novel system to facilitate the rapid and easy cloning of multiple genes (>10) in under a week. Using this system we have been able to successfully clone, overexpress, and purify a number of large multimeric proteins from insect cells, including the chromatin remodeling complexes SWR1 and INO80. Using Förster resonance energy transfer (FRET)-based assays we have demonstrated that our overexpressed enzymes have activities comparable to those purified from sources where the proteins are expressed under their endogenous promoters.

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http://dx.doi.org/10.1016/bs.mie.2017.03.015DOI Listing

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