Abstract
The formation of protein complexes through direct protein–protein interaction is essential for virtually every biological process, and accordingly the ability to determine the interaction properties of specific proteins is important to understand these processes. Förster resonance energy transfer (FRET) measurements are state-of-the-art confocal fluorescence microscopy- and imaging-based techniques that allow the analysis of protein interactions in vivo and in planta, in specific compartments of single cells or tissues. Here we provide a step-by-step guide to perform FRET measurements by acceptor photobleaching (APB) and fluorescence lifetime imaging microscopy (FLIM) in the plant expression system Nicotiana benthamiana.
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Acknowledgments
M. S. and R. S. thank Petra Žádníková and Maike Breiden for helpful comments and discussion of the manuscript and the CAi of the HHU for technical support. This work was funded by the D. F. G. through the Exc1028 (CEPLAS).
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Somssich, M., Simon, R. (2017). Studying Protein–Protein Interactions In Planta Using Advanced Fluorescence Microscopy. In: Busch, W. (eds) Plant Genomics. Methods in Molecular Biology, vol 1610. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7003-2_17
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DOI: https://doi.org/10.1007/978-1-4939-7003-2_17
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