Abstract: The invention relates generally to methods for preparing recombinant nucleosomes. In particular, the invention relates to methods for ligating a histone peptide onto a fully assembled recombinant nucleosome. The invention further relates to modified core histone proteins, histone peptides to be ligated to the modified core histone proteins, and fully assembled recombinant nucleosomes and libraries of recombinant nucleosomes prepared by the methods of the invention.

Abstract: The invention relates to assays for nucleosome remodeling activity using functionalized recombinant mononucleosomes. The functionalized recombinant mononucleosomes comprise a histone octamer comprising recombinant histone H2A, H2B, H3 and H4 proteins and a DNA template comprising a nucleosome positioning sequence that effectively positions the histone octamer and a signal site. The invention further relates to methods of using the assay to quantify enzymatic activity of remodeling enzymes and identifying modulators of remodeling enzyme activity.

Abstract: The invention relates to the use of recombinant/semi-synthetic nucleosomes carrying histone and/or DNA modifications as a reference standard for quantification of covalently modified (on the histone proteins or wrapping DNA), variant, or mutant nucleosomes (collectively “modified nucleosomes” or “nucleosome modifications”) from a biological sample. The invention further relates to methods of using the assay to accurately quantify single or combinatorial nucleosome modifications as biomarkers of disease.