Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024:2724:211-223.
doi: 10.1007/978-1-0716-3485-1_15.

Bimolecular Fluorescence Complementation (BiFC) in Host-Virus Interactions

Fredy Davi Albuquerque Silva  1   2 , João Paulo Batista Machado  3   4 , Pedro Augusto Braga Dos Reis  5   6
Affiliations

Bimolecular Fluorescence Complementation (BiFC) in Host-Virus Interactions

Fredy Davi Albuquerque Silva et al. Methods Mol Biol. 2024.

Abstract

Bimolecular fluorescence complementation (BiFC) is an assay widely used for studying protein-protein interactions and determining the subcellular localization of proteins. This technique involves fusing the proteins of interest to separate structural domains of a fluorescent protein, followed by transient expression in cells. The interaction between the proteins of interest in vivo allows the reconstitution of the fluorescence that can be visualized by fluorescence microscopy. BiFC has been particularly useful in investigating the interactions between viral and host proteins. Here, we describe the steps involved in preparing expression cassettes that allow the expression of proteins of interest fused to nonfluorescent fragments of yellow fluorescent protein (YFP), Agrobacterium transformations, and agroinfiltration of Nicotiana benthamiana leaves to facilitate virus protein-host protein interactions. Finally, high-resolution images can be obtained by analyzing the leaves under a confocal microscope.

Keywords: Agroinfiltration; BiFC; Confocal microscopy; Viral protein–host protein interactions.

PubMed Disclaimer

References

    1. Kodama Y, Hu CD (2012) Bimolecular fluorescence complementation (BiFC): a 5-year update and future perspectives. BioTechniques 53(5):285–298. https://doi.org/10.2144/000113943 - DOI
    1. Citovsky V, Lee LY, Vyas S, Glick E, Chen MH, Vainstein A, Gafni Y, Gelvin SB, Tzfira T (2006) Subcellular localization of interacting proteins by bimolecular fluorescence complementation in planta. J Mol Biol 362(5):1120–1131. https://doi.org/10.1016/j.jmb.2006.08.017 - DOI
    1. Hyodo K, Okuno T (2020) Hijacking of host cellular components as proviral factors by plant-infecting viruses. Adv Virus Res 107:37–86. https://doi.org/10.1016/bs.aivir.2020.04.002 - DOI
    1. Silva FDA, Raimundo GS, Fontes EPB (2022) Begomovirus–host protein-protein interactions in intracellular virus movement. In: Gaur RK, Sharma P, Czosnek H (eds) Geminivirus: Detection, diagnosis and management. Academic, pp 347–356. https://doi.org/10.1016/B978-0-323-90587-9.00023-7 - DOI
    1. Brustolini OJB, Machado JPB, Condori-Apfata JA, Coco D, Deguchi M, Loriato VAP, Pereira WA, Alfenas-Zerbini P, Zerbini FM, Inoue-Nagata AK, Santos AA, Chory J, Silva FF, Fontes EPB (2015) Sustained NIK-mediated antiviral signalling confers broad-spectrum tolerance to begomoviruses in cultivated plants. Plant Biotechnol J 13(9):1300–1311. https://doi.org/10.1111/pbi.12349 - DOI

Publication types

Substances

LinkOut - more resources