Production and Assay of Recombinant Multisubunit Chromatin Remodeling Complexes.

Autor:	Rees DM; Section of Structural Biology, Imperial College London, London, United Kingdom., Willhoft O; Section of Structural Biology, Imperial College London, London, United Kingdom., Lin CL; Section of Structural Biology, Imperial College London, London, United Kingdom., Bythell-Douglas R; Section of Structural Biology, Imperial College London, London, United Kingdom., Wigley DB; Section of Structural Biology, Imperial College London, London, United Kingdom. Electronic address: d.wigley@imperial.ac.uk.
Jazyk:	angličtina
Zdroj:	Methods in enzymology [Methods Enzymol] 2017; Vol. 592, pp. 27-47. Date of Electronic Publication: 2017 Apr 17.
DOI:	10.1016/bs.mie.2017.03.015
Abstrakt:	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. (© 2017 Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu