Protein fragment bimolecular fluorescence complementation analyses for the in vivo study of protein-protein interactions and cellular protein complex localizations.
The analyses of protein-protein interactions are crucial for understanding cellular processes including signal transduction, protein trafficking, and movement. Protein fragment complementation assays are based on the reconstitution of protein function when non-active protein fragments are brought together by interacting proteins that were genetically fused to these protein fragments. Bimolecular fluorescence complementation (BiFC) relies on the reconstitution of fluorescent proteins and enables both the analysis of protein-protein interactions and the visualization of protein complex formations in vivo. Transient expression of proteins is a convenient approach to study protein functions in planta or in other organisms and minimizes the need for time-consuming generation of stably expressing transgenic organisms. Here we describe protocols for BiFC analyses in Nicotiana benthamiana and Arabidopsis thaliana leaves transiently transformed by Agrobacterium infiltration. Further, we discuss different BiFC applications and provide examples for proper BiFC analyses in planta.
Division of Biological Sciences, Cell and Developmental Biology Section and Center for Food and Fuel for the 21st Century, University of California San Diego, La Jolla, CA, USA.
Rainer
R
Waadt
Kathrin
K
Schlücking
Julian I
JI
Schroeder
Jörg
J
Kudla
D060054:Agrobacterium; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D010446:Peptide Fragments / Q000737:chemistry*; D025941:Protein Interaction Mapping / Q000379:methods*; D021381:Protein Transport; D012639:Seeds / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000378:metabolism; D014162:Transfection
D029681:Arabidopsis Proteins; D010446:Peptide Fragments; D049452:Green Fluorescent Proteins