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Methods Mol Biol. 2017;1610:267-285. doi: 10.1007/978-1-4939-7003-2_17.

Studying Protein-Protein Interactions In Planta Using Advanced Fluorescence Microscopy.

Author information

1
Institute for Developmental Genetics, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany.
2
School of Biosciences, University of Melbourne, Melbourne, VIC, Australia.
3
Institute for Developmental Genetics, Cluster of Excellence on Plant Sciences (CEPLAS), and Center for Advanced Imaging (CAi), Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany. ruediger.simon@hhu.de.

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.

KEYWORDS:

APB; Acceptor photobleaching; Confocal fluorescence microscopy; FLIM; FRET; Fluorescence lifetime; Live cell imaging; Nicotiana benthamiana; Protein–protein interaction

PMID:
28439869
DOI:
10.1007/978-1-4939-7003-2_17
[Indexed for MEDLINE]

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