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一种新的饱和诱变方法: 利用 Phosphorothioates 对 RNA 调控蛋白结合位点的单步表征
JoVE Journal
Genetica
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JoVE Journal Genetica
A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates
DOI:

11:49 min

August 21, 2018

Capitoli

  • 00:04Titolo
  • 00:30Generation of a Library of Mutants – Doping DNA Template with Non-wild Type Nucleotides
  • 01:20Synthesis of RNA
  • 05:24Protein Binding Reaction and Separation of Bound RNA
  • 08:05Analysis of Iodine-cleaved Phosphorothioate Products for Detection of Mutant Nucleotide Positions
  • 09:07Results: Characterization of Protein Binding Site in RNA
  • 10:22Conclusion

Summary

Traduzione automatica

结合特定 RNA 序列的蛋白质在基因表达中扮演重要角色。对这些结合部位的详细描述对于我们理解基因调控至关重要。本文介绍了 RNA 蛋白质结合部位饱和诱变的单步法方法。这种方法与 RNA 中的所有蛋白结合部位有关。

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