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NEMO IKK绑定域的生产、结晶和结构确定
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO
DOI:

13:02 min

December 28, 2019

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Chapters

  • 00:04Title
  • 01:12Large Scale Expression of Histidine-6 (His6)-Tagged Nuclear Factor-kappa-B Essential Modulator Crystallography Construct (NEMO-EEAA)
  • 02:33His6-Tagged NEMO-EEAA Purification
  • 04:42His6 Tag Cleavage and Purification
  • 07:09Sparse Matrix Screening
  • 08:23Seed Stock Generation
  • 09:00Crystal Looping
  • 09:43X-Ray Data Processing and Structure Solution
  • 10:38Results: Representative Production, Crystallization, and Structure Determination of the IKK-Binding Domain of NEMO
  • 12:28Conclusion

Summary

Automatic Translation

我们描述了通过X射线晶体学测定NEMOIKK结合域的结构确定方案。这些方法包括蛋白质表达、纯化和表征,以及成功优化晶体和确定无结合形式蛋白质的结构的策略。

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