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
JoVE Journal
Biologia
Author Produced
É necessária uma assinatura da JoVE para visualizar este conteúdo.  Faça login ou comece sua avaliação gratuita.
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

Determination of Tripartite Interaction between Two Monomers of a MADS-box Transcription Factor and a Calcium Sensor Protein by BiFC-FRET-FLIM Assay

DOI:
10.3791/62791-v

•

14:34 min

•

December 25, 2021

•

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

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

Tadução automática

Here we present, a method to visualize ternary complex formation between three protein partners using fluorescent-tagged proteins by BiFC based FRET-FLIM assay. This method is valuable for studying protein-protein interaction complexes in vivo.

Tags

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

Article

Embed

ADICIONAR À PLAYLIST

Usage Statistics

Vídeos Relacionados

check_url/pt/62791?article_type=v

Generation of Induced Pluripotent Stem Cells by Reprogramming Mouse Embryonic Fibroblasts with a Four Transcription Factor, Doxycycline Inducible Lentiviral Transduction System
check_url/pt/62791?article_type=v

Bimolecular Fluorescence Complementation (BiFC) Assay for Protein-Protein Interaction in Onion Cells Using the Helios Gene Gun
check_url/pt/62791?article_type=v

Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor
check_url/pt/62791?article_type=v

Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro
check_url/pt/62791?article_type=v

A Liquid Phase Affinity Capture Assay Using Magnetic Beads to Study Protein-Protein Interaction: The Poliovirus-Nanobody Example
check_url/pt/62791?article_type=v

Protein Purification-free Method of Binding Affinity Determination by Microscale Thermophoresis
check_url/pt/62791?article_type=v

A Sensitive and Specific Quantitation Method for Determination of Serum Cardiac Myosin Binding Protein-C by Electrochemiluminescence Immunoassay
check_url/pt/62791?article_type=v

Characterization of G Protein-coupled Receptors by a Fluorescence-based Calcium Mobilization Assay
check_url/pt/62791?article_type=v

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
check_url/pt/62791?article_type=v

Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay (PCA) in Living Cells

Read Article