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PCR 突变体、克隆、表达、快速蛋白质纯化协议和色氨酸合成酶野生类型和突变形式的结晶
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JoVE Journal Biochimica
PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase
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PCR 突变体、克隆、表达、快速蛋白质纯化协议和色氨酸合成酶野生类型和突变形式的结晶

DOI:
10.3791/61839-v

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09:31 min

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September 26, 2020

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, , ,
1Department of Chemistry,University of California-Riverside, 2Department of Biochemistry,University of California-Riverside

Capitoli

  • 00:04Introduction
  • 00:44Purification of Wild Type or Mutant Form of α2β2 StTS Complex
  • 03:53Optimized Crystallization for Wild Type and Mutant Form of the α2β2 StTS Comple
  • 06:49Results: Purification of the α- and β -Subunits of the Tryptophan Synthase and the Wild Type and Mutant α2β2 StTS Complexes
  • 08:50Conclusion

Summary

Traduzione automatica

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本文介绍了一系列连续的方法来表达和净化 沙门氏菌色氨酸 合成酶，使该协议成为一天中净化蛋白质复合物的快速系统。覆盖的方法包括现场定向突变、 大肠杆菌中的蛋白质表达、亲和色谱、凝胶过滤色谱和结晶。

Tags

Keywords: PCR MutagenesisCloningExpressionProtein PurificationTryptophan SynthaseSalmonella TyphimuriumAmmonium Sulfate FractionationSize Exclusion ChromatographySDS-PAGEProtein ConcentrationCrystallization

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