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Production and Assay of Recombinant Multisubunit Chromatin Remodeling Complexes.

Methods Enzymol.. 2017-04; 
Rees DM,Willhoft O,Lin CL,Bythell-Douglas R,Wigley DB.
Products/Services Used Details Operation
Codon Optimization ... complexes. 2. Cloning. Genes for all proteins were codon optimized and then synthesized (GenArt, Thermo Fisher Scientific, Waltham, MA or GenScript, Piscataway, NJ). The advantages of getting them synthesized were twofold. … Get A Quote

摘要

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.

关键词

Chromatin remodeling; DNA repair; Histone exchange; Insect cell expression; Multisubunit complexes; Nuclear actin