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Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System
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Biologia
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JoVE Journal Biologia
Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System

Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System

DOI:
10.3791/3473-v

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08:21 min

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September 16, 2011

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, , , ,
1Department of Biology,University of Western Ontario, 2Southern Crop Protection and Food Research Centre,Agriculture and Agri-Food Canada

Capítulos

  • 00:05Título
  • 01:16Preparation of Agrobacterium culture for infiltration
  • 04:06Infiltration
  • 06:03Observation of the reconstituted YFP signal
  • 07:40Conclusion

Summary

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We have developed a technique to test protein-protein interactions in plant. A yellow fluorescent protein (YFP) is split into two non-overlapping fragments. Each fragment is cloned in-frame to a gene of interest via Gateway system, enabling expression of fusion proteins. Reconstitution of YFP signal only occurs when the inquest proteins interact.

Tags

Protein InteractionsPlantGateway CompatibleBimolecular Fluorescence ComplementationBiFC SystemYellow Fluorescent ProteinIn Vivo Interaction TestingNicotiana BenthamianaAgrobacterium Mediated TransformationFusion ProteinsYFP Signal ReconstitutionYeast Two Hybrid AssayIndirect InteractionsGateway TechnologyGene Of Interest (GOI)Expression And Functional Analysis SystemsOrientation And Reading Frame MaintenanceExpression-ready ClonesBiFC And Y2H VectorsEase Of Use

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