Detection of protein-protein interactions in plants using bimolecular fluorescence complementation

Keren Bracha-Drori, Keren Shichrur, Aviva Katz, Moran Oliva, Ruthie Angelovici, Shaul Yalovsky*, Nir Ohad

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

342 Scopus citations


Protein function is often mediated via formation of stable or transient complexes. Here we report the determination of protein-protein interactions in plants using bimolecular fluorescence complementation (BiFC). The yellow fluorescent protein (YFP) was split into two non-overlapping N-terminal (YN) and C-terminal (YC) fragments. Each fragment was cloned in-frame to a gene of interest, enabling expression of fusion proteins. To demonstrate the feasibility of BiFC in plants, two pairs of interacting proteins were utilized: (i) the α and β subunits of the Arabidopsis protein farnesyltransferase (PFT), and (ii) the polycomb proteins, FERTILIZATION-INDEPENDENT ENDOSPERM (FIE) and MEDEA (MEA). Members of each protein pair were transiently co-expressed in leaf epidermal cells of Nicotiana benthamiana or Arabidopsis. Reconstitution of a fluorescing YFP chromophore occurred only when the inquest proteins interacted. No fluorescence was detected following co-expression of free non-fused YN and YC or non-interacting protein pairs. Yellow fluorescence was detected in the cytoplasm of cells that expressed PFT α and β subunits, or in nuclei and cytoplasm of cells that expressed FIE and MEA. In vivo measurements of fluorescence spectra emitted from reconstituted YFPs were identical to that of a non-split YFP, confirming reconstitution of the chromophore. Expression of the inquest proteins was verified by immunoblot analysis using monoclonal antibodies directed against tags within the hybrid proteins. In addition, protein interactions were confirmed by immunoprecipitations. These results demonstrate that plant BiFC is a simple, reliable and relatively fast method for determining protein-protein interactions in plants.

Original languageEnglish
Pages (from-to)419-427
Number of pages9
JournalPlant Journal
Issue number3
StatePublished - Nov 2004


  • Bimolecular fluorescence complementation
  • Polycomb group proteins
  • Protein farnesyltransferase
  • Protein-protein interaction


Dive into the research topics of 'Detection of protein-protein interactions in plants using bimolecular fluorescence complementation'. Together they form a unique fingerprint.

Cite this