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在大肠杆菌中生成特异的乙酰蛋白的简易协议
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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
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在大肠杆菌中生成特异的乙酰蛋白的简易协议

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

基因编码扩展是研究广泛的生物过程, 包括蛋白质乙酰化的有力工具。在这里, 我们演示了一个轻便的协议, 利用这种技术, 在特定的地点, 在大肠杆菌细胞生成均匀乙酰蛋白。

Tags

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

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