Author: Ramasubramanian Sundaramoorthy; Amanda L. Hughes; Hassane El-Mkami; David Norman; Tom Owen-Hughes
Title: Structure of the chromatin remodelling enzyme Chd1 bound to a ubiquitinylated nucleosome Document date: 2018_3_30
ID: ct2zhauz_49
Snippet: Recombinant expression of xH2B K120C and His-TEV-Ubiquitin G76C mutant proteins was induced with IPTG for 4 hours in Rosetta 2 DE3 pLysS cells grown . CC-BY-NC 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/290874 doi: bioRxiv preprint at 37â°C. Inclusion body purification followed by cation exchange chromatography was per.....
Document: Recombinant expression of xH2B K120C and His-TEV-Ubiquitin G76C mutant proteins was induced with IPTG for 4 hours in Rosetta 2 DE3 pLysS cells grown . CC-BY-NC 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/290874 doi: bioRxiv preprint at 37â°C. Inclusion body purification followed by cation exchange chromatography was performed to isolate the histone protein. Ubiquitin was purified using HisPur cobalt resin with 150mM sodium chloride/20mM Tris pH8 buffer and eluted with 350mM imidazole. Histones and ubiquitin were desalted by dialysis into water with 2mM 2-mercaptoethanol and lyophilised for storage.
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