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Split-Luciferase Complementation for Analysis of Virus–Host Protein Interactions

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Plant Virology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2400))

Abstract

Productive viral infection entails highly regulated and sequential protein–protein interactions between viral factors and between virus and host factors. Deciphering such interactions is of paramount importance for a better understanding of virus infection cycles and the development of new strategies for virus prevention and control. In this protocol, we describe a split-luciferase complementation (SLC ) assay for the detection of protein–protein interaction in Nicotiana benthamiana leaves following agroinfiltration-mediated transient protein expression. In this assay, the firefly luciferase protein is divided into two halves, each expressed as a fusion to a prey or bait protein, respectively. Interaction of the two candidate proteins brings the two otherwise nonfunctional halves into close proximity to restore the luciferase activity, which catalyzes the substrate D-luciferin to emit luminescence. The SLC assay allows for noninvasive, quantitative measurement of dynamic protein interactions in living cells within their native cellular compartments.

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Acknowledgments

This work was supported in part by grants from the Natural Science Foundation of Zhejiang Province, China (grant no. LZ20C140004), and the National Natural Science Foundation of China (grant no. 31671996) to Z.L.

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Authors and Affiliations

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

    Yan Liang & Zhenghe Li

  2. Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insect Pests, Zhejiang University, Hangzhou, China

    Zhenghe Li

  3. Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, Zhejiang University, Hangzhou, China

    Zhenghe Li

Authors
  1. Yan Liang
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  2. Zhenghe Li
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Corresponding author

Correspondence to Zhenghe Li .

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Editors and Affiliations

  1. London Research and Development Centre, Agriculture and Agri-Food Canada, London, ON, Canada

    Aiming Wang

  2. State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China

    Yi Li

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Liang, Y., Li, Z. (2022). Split-Luciferase Complementation for Analysis of Virus–Host Protein Interactions. In: Wang, A., Li, Y. (eds) Plant Virology . Methods in Molecular Biology, vol 2400. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1835-6_6

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  • DOI: https://doi.org/10.1007/978-1-0716-1835-6_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1834-9

  • Online ISBN: 978-1-0716-1835-6

  • eBook Packages: Springer Protocols

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