Site Specific Lysine Acetylation of Histones for Nucleosome Reconstitution using Genetic Code Expansion in Escherichia coli

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Biochemistry

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JoVE Journal Biochemistry

Site Specific Lysine Acetylation of Histones for Nucleosome Reconstitution using Genetic Code Expansion in Escherichia coli

Article DOI: 10.3791/62113-v • 07:26 min • December 26th, 2020

December 26th, 2020 •

Here we present a method to express acetylated histone proteins using genetic code expansion and assemble reconstituted nucleosomes in vitro.

Site-specific Lysine Acetylation Histones Nucleosome Reconstitution Genetic Code Expansion Escherichia Coli Modified Mononucleosome Epigenetic Experiments Acetylated Reconstituted Mononucleosomes Biochemical Assays Binding Assays Structure Determination Versatility 2YT Media Antibiotics Co-transformed Cell Stock Tag-containing Plasmid PEVOL-AckRS OD (optical Density) Inducing Agents AcK (acetyllysine) IPTG (isopropyl Î²-D-1-thiogalactopyranoside) Arabinose Minus 80 Degrees Celsius Histone Lysis Buffer Sonicate