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AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants
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AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants
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AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants

DOI:
10.3791/64428-v

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08:36 min

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September 16, 2022

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, , , ,
1College of Bioscience and Biotechnology,Yangzhou University

Chapters

  • 00:00Introduction
  • 00:42Preparation of Plant Material
  • 01:14Agroinfiltration
  • 03:19Collection of Samples and Total Protein Extraction from Leaf
  • 04:18Equilibrating the Desalting Column
  • 04:49Magnetic Beads Washing and Enrichment of Biotinylated Proteins
  • 06:56Results: AirID-Based Proximity Labeling in Cucumis sativus
  • 07:50Conclusion

Summary

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Automatic Translation

Here, we present a step-by-step protocol for performing the proximity labeling (PL) experiment in cucumber (Cucumis sativus L.) using AT4G18020 (APRR2)-AirID protein as a model. The method describes the construction of a vector, the transformation of a construct through agroinfiltration, biotin infiltration, protein extraction, and purification of biotin-labeled proteins through affinity purification technique.

Tags

Keywords: AirIDProtein-protein InteractionProximity LabelingCucumis SativusAgrobacterium TumefaciensAgroinfiltrationAPRR2-AirIDPlant BiotechnologyMolecular BiologyProtein Analysis

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