Expression, purification and proteomic analysis of recombinant histone H 4 acetylated at lysine 16

Many histone covalent modifications have been identified and shown to play key regulatory roles in eukaryotic transcription, DNA damage repair, and replication. In vitro experiments designed to understand the mechanistic role of individual modifications require the availability of substantial quantities of pure histones, homogeneously modified at specific residues. We have applied the amber stop codon/suppressor t RNA strategy to the production of histone H 4 acetylated at lysine 16, a particularly important isoform of this histone. Our success relies on adapting the H 4 DNA sequence to the codon preference of E . coli and on preventing the premature decay of the H 4 m RNA . These modifications to the original procedure render it easily applicable to the generation of any covalently modified histone H 4 isoform.