Utilizing bimolecular fluorescence complementation (BiFC) to assay protein-protein interaction in plants.
Protein function is often mediated by the formation of stable or transient complexes. Here we present a method for testing protein-protein interactions in plants designated bimolecular fluorescence complementation (BiFC). The advantages of BiFC are its simplicity, reliability, and the ability to observe protein-protein interactions in different cellular compartments including membranes. BiFC is based on splitting the yellow fluorescent protein (YFP) into two nonoverlapping N-terminal (YN) and C-terminal (YC) fragments. Each fragment is cloned in-frame with a gene of interest, enabling expression of a fusion protein. Reconstitution of the fluorescing YFP chromophore takes place upon interaction of protein pairs that are coexpressed in the same cells.
Ohad
Nir
N
Department of Plant Sciences, Tel-Aviv University, Tel-Aviv, Israel.
Yalovsky
Shaul
S
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D005453:Fluorescence; D015870:Gene Expression; D005822:Genetic Vectors; D015151:Immunoblotting / Q000379:methods; D047468:Immunoprecipitation / Q000379:methods; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods; D018515:Plant Leaves / Q000235:genetics / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010944:Plants / Q000235:genetics / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods*; D012231:Rhizobium / Q000235:genetics
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D010940:Plant Proteins; C054562:yellow fluorescent protein, Bacteria