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Bestemmelse av trepartsinteraksjon mellom to monomerer av en MADS-boks transkripsjonsfaktor og et kalsiumsensorprotein av BiFC-FRET-FLIM Assay
JoVE Journal
Biology
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JoVE Journal Biology
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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Chapters

  • 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

Automatic Translation

Her presenterer vi en metode for å visualisere ternær kompleks formasjon mellom tre proteinpartnere ved hjelp av fluorescerende merkede proteiner av BiFC-basert FRET-FLIM-analyse. Denne metoden er verdifull for å studere proteinproteininteraksjonskomplekser in vivo.

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