Abstract
The bimolecular fluorescence complementation (BiFC) assay is a versatile technique for investigating protein-protein interaction (PPI) in living systems. The BiFC assay exploits the color-emitting moiety and the modular structure of fluorescent proteins to provide both temporal and spatial information of the PPI. The modular property of fluorescent proteins enables researchers to strategically partition a fluorescent protein into two nonfluorescent units, which can be independently fused to other proteins. When the fusion proteins interact with each other, the nonfluorescent fragments reconstitute to generate a fluorescence signal. PPI can then be detected by capturing the fluorescence signal with a fluorescence microscope. In this chapter, the Venus fluorescent protein is employed to demonstrate the application of the BiFC assay.
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Acknowledgements
I would like to thank Drs. Haian Fu and Jonathan Havel for their generous gifts of the pSCM167-NV and pDEST26-CV Gateway® destination vectors as well as constructive input to make this assay work.
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Pham, C.D. (2015). Detection of Protein-Protein Interaction Using Bimolecular Fluorescence Complementation Assay. In: Meyerkord, C., Fu, H. (eds) Protein-Protein Interactions. Methods in Molecular Biology, vol 1278. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2425-7_32
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DOI: https://doi.org/10.1007/978-1-4939-2425-7_32
Publisher Name: Humana Press, New York, NY
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