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A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli
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Immunology and Infection
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JoVE Journal Immunology and Infection
A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli

A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli

DOI:
10.3791/57061-v

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11:08 min

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December 09, 2017

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, , , ,
1Department of Chemistry and Biochemistry,University of Arkansas, 2Cell and Molecular Biology Program,University of Arkansas, 3Department of Biological Sciences,University of Arkansas, 4Fayetteville High School

Chapters

  • 00:00Title
  • 00:09Introduction
  • 01:03Protocol
  • 01:07Introducing the Stop Codon into the Target Gene
  • 02:58Expressing and Purifying Site-specifically Acetylated Protein
  • 06:21Biochemically Characterizing Acetylated Protein
  • 09:39Results
  • 10:37Summary

Summary

Automatic Translation

Automatic Translation

Genetic code expansion serves as a powerful tool to study a wide range of biological processes, including protein acetylation. Here we demonstrate a facile protocol to exploit this technique for generating homogeneously acetylated proteins at specific sites in Escherichia coli cells.

Tags

Keywords: Site-specific AcetylationProtein AcetylationGenetic Code ExpansionPyrrolysyl-tRNA SynthetaseAmber Stop CodonE. Coli ExpressionSite-directed MutagenesisAcetyllysine Incorporation

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