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Bestemmelse af trepartsinteraktion mellem to monomerer af en MADS-box transskriptionsfaktor og et calciumsensorprotein ved BiFC-FRET-FLIM Assay
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Biologie
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Journal JoVE Biologie
Determination of Tripartite Interaction between Two Monomers of a MADS-box Transcription Factor and a Calcium Sensor Protein by BiFC-FRET-FLIM Assay

Bestemmelse af trepartsinteraktion mellem to monomerer af en MADS-box transskriptionsfaktor og et calciumsensorprotein ved BiFC-FRET-FLIM Assay

DOI:
10.3791/62791-v

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14:34 min

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December 25, 2021

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, , , , , , ,
1Interdisciplinary Centre for Plant Genomics, Department of Plant Molecular Biology,University of Delhi South Campus, 2University School of Biotechnology,Guru Gobind Singh Indraprastha University

Chapitres

  • 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

Traduction automatique

Traduction automatique

Her præsenterer vi, en metode til at visualisere ternary kompleks dannelse mellem tre proteinpartnere ved hjælp af fluorescerende-tagged proteiner ved BiFC baseret FRET-FLIM assay. Denne metode er værdifuld til undersøgelse af protein-protein interaktion komplekser in vivo.

Tags

Protein-protein InteractionsBiFCFRETFLIMTripartite InteractionMADS-box Transcription FactorCalcium Sensor ProteinHomodimerizationNicotiana BenthamianaAgroinfiltration

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