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JoVE Journal
Biochemistry

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

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

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Summary

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

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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 β-D-1-thiogalactopyranoside) Arabinose Minus 80 Degrees Celsius Histone Lysis Buffer Sonicate
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