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通过BiFC-FRET-FLIM测定MADS盒转录因子和钙传感器蛋白两种单体之间的三方相互作用
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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
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通过BiFC-FRET-FLIM测定MADS盒转录因子和钙传感器蛋白两种单体之间的三方相互作用

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

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

在这里，我们提出了一种通过基于BiFC的FRET-FLIM测定使用荧光标记蛋白可视化三元复合物形成的方法。该方法对于研究 体内蛋白质 - 蛋白质相互作用复合物很有价值。

Tags

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

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