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BIFC-FRET-FLIMアッセイによるMADS型転写因子の2つのモノマーとカルシウムセンサタンパク質の三者間相互作用の測定
JoVE Journal
Biologia
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JoVE Journal Biologia
Determination of Tripartite Interaction between Two Monomers of a MADS-box Transcription Factor and a Calcium Sensor Protein by BiFC-FRET-FLIM Assay
DOI:

14:34 min

December 25, 2021

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Capítulos

  • 00:08Introduction
  • 01:17Cloning of Genes in Entry and Destination Vectors
  • 02:01Growth Conditions for Nicotiana benthamiana Plants
  • 03:05Prepare Bacterial Strains for Agroinfiltration
  • 06:20Prepare Slides for Fluorescence Visualization
  • 06:45Fret-Flim Analysis Using a Confocal Laser Scanning Microscope
  • 10:51Fret-Flim Analysis Using a Confocal Laser Scanning Microscope: The Tripartite Interaction
  • 12:31Representative Results
  • 13:42Conclusion

Summary

Tadução automática

ここでは、BiFCベースのFRET-FLIMアッセイによる蛍光タグ付きタンパク質を用いた3つのタンパク質パートナー間の三元複合体形成を可視化する方法を提示する。この方法は、 生体内のタンパク質とタンパク質相互作用複合体を研究する上で有用である。

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