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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

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

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3,837 Views

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07:26 min

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December 26, 2020

DOI:

10.3791/62113-v

DOI:
10.3791/62113-v

•

07:26 min
December 26, 2020

•

4 Views
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1Department of Chemistry,Texas A&M University

Summary

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

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Lysine AcetylationHistonesNucleosome ReconstitutionGenetic Code ExpansionEscherichia ColiMononucleosomeEpigenetic ExperimentsBiochemical AssaysBinding AssaysStructure DeterminationHistone Lysis BufferInclusion BodiesNickel-NTA PurificationAcetylated Histone ProteinDialysisAcetic Acid Buffer

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Rowlett, C. M., Liu, W. R. Site Specific Lysine Acetylation of Histones for Nucleosome Reconstitution using Genetic Code Expansion in Escherichia coli. J. Vis. Exp. (166), e62113, doi:10.3791/62113 (2020).

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