Production and Assay of Recombinant Multisubunit Chromatin Remodeling Complexes

Abstract 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 Forster 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.