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Related Experiment Video

Updated: Jul 10, 2025

Luciferase Complementation Imaging Assay in Nicotiana benthamiana Leaves for Transiently Determining Protein-protein Interaction Dynamics
07:55

Luciferase Complementation Imaging Assay in Nicotiana benthamiana Leaves for Transiently Determining Protein-protein Interaction Dynamics

Published on: November 20, 2017

14.0K

Split-Luciferase Complementation Imaging Assay in Virus-Plant Interactions.

Huang Tan1, Xi Zhang2,3, Rosa Lozano-Duran4

  • 1Department of Plant Biochemistry, Centre for Plant Molecular Biology (ZMBP), Eberhard Karls University Tübingen, Tübingen, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|November 21, 2023
PubMed
Summary

This study demonstrates a simple, fast, and inexpensive split-luciferase assay to visualize plant-virus protein interactions in vivo. The method effectively tracks interactions between viral proteins and host factors during infection.

Keywords:
In vivoNicotiana benthamianaProtein−protein interactionSplit-luciferase complementation assayViral protein

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Area of Science:

  • Plant Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Protein-protein interactions are key to viral infection outcomes.
  • Studying viral-host protein interactions in plants is essential.
  • Existing methods for studying these interactions can be complex.

Purpose of the Study:

  • To present a simple, fast, quantitative, and inexpensive method for studying protein-protein interactions in plants.
  • To demonstrate the utility of the split-luciferase complementation imaging assay for in vivo analysis of viral-host interactions.
  • To provide a protocol for testing protein-protein interactions using transient Agrobacterium tumefaciens-mediated transformation.

Main Methods:

  • Split-luciferase complementation imaging assay.
  • Transient Agrobacterium tumefaciens-mediated transformation.
  • In vivo detection of protein-protein interactions in Nicotiana benthamiana leaves.

Main Results:

  • The split-luciferase assay successfully detected the interaction between Tomato Yellow Leaf Curl Virus (TYLCV) V2 protein and Nicotiana benthamiana Argonaute 4 (AGO4) in vivo.
  • The assay provides a quantitative and rapid assessment of protein-protein interactions.
  • The method is adaptable for studying various viral-host protein interactions.

Conclusions:

  • The split-luciferase complementation imaging assay is a powerful and accessible tool for studying plant-virus protein interactions.
  • This method facilitates in vivo analysis of interactions within the context of infection.
  • The presented protocol offers a straightforward approach for researchers to investigate protein-protein interactions in plants.