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Related Experiment Video

Updated: Oct 10, 2025

Demonstration of Heterologous Complexes formed by Golgi-Resident Type III Membrane Proteins using Split Luciferase Complementation Assay
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Split-Luciferase Complementation for Analysis of Virus-Host Protein Interactions.

Yan Liang1, Zhenghe Li2,3,4

  • 1State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou, China.

Methods in Molecular Biology (Clifton, N.J.)
|December 14, 2021
PubMed
Summary

This study introduces a split-luciferase complementation (SLC) assay to detect protein-protein interactions in plants. This method enables noninvasive, quantitative monitoring of dynamic interactions crucial for understanding viral infections.

Keywords:
AgroinfiltrationLuciferinNicotiana benthamianaProtein–protein interactionSplit-luciferase complementation assay

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

  • Molecular Biology
  • Virology
  • Plant Science

Background:

  • Productive viral infections rely on intricate protein-protein interactions between viral and host factors.
  • Understanding these interactions is key to deciphering viral infection cycles and developing antiviral strategies.

Purpose of the Study:

  • To describe a novel split-luciferase complementation (SLC) assay for detecting protein-protein interactions.
  • To enable noninvasive, quantitative measurement of dynamic protein interactions in living plant cells.

Main Methods:

  • The split-luciferase complementation (SLC) assay utilizes agroinfiltration-mediated transient expression in Nicotiana benthamiana leaves.
  • Firefly luciferase is split into two halves, fused to bait and prey proteins.
  • Protein interaction reconstitutes luciferase activity, enabling luminescence detection upon D-luciferin addition.

Main Results:

  • The SLC assay successfully detects protein-protein interactions in living plant cells.
  • The method allows for quantitative and noninvasive measurement of these interactions.
  • Interactions are visualized within the native cellular compartments of the plant.

Conclusions:

  • The described SLC assay provides a powerful tool for studying viral and host protein interactions in plants.
  • This technique facilitates a deeper understanding of virus infection mechanisms.
  • It offers a valuable approach for the development of novel antiviral interventions.