Bimolecular fluorescence complementation as a tool to study interactions of regulatory proteins in plant protoplasts

Sitakanta Pattanaik, Joshua R Werkman, Ling Yuan

Research output: Contribution to journalArticlepeer-review


Protein-protein interactions are an important aspect of the gene regulation process. The expression of a gene in response to certain stimuli, within a specific cell type or at a particular developmental stage, involves a complex network of interactions between different regulatory proteins and the cis-regulatory elements present in the promoter of the gene. A number of methods have been developed to study protein-protein interactions in vitro and in vivo in plant cells, one of which is bimolecular fluorescence complementation (BiFC). BiFC is a relatively simple technique based upon the reconstitution of a fluorescent protein. The interacting protein complex can be visualized directly in a living plant cell when two non-fluorescent fragments, of an otherwise fluorescent protein, are fused to proteins found within that complex. Interaction of tagged proteins brings the two non-fluorescent fragments into close proximity and reconstitutes the fluorescent protein. In addition, the subcellular location of an interacting protein complex in the cell can be simultaneously determined. Using this approach, we have successfully demonstrated a protein-protein interaction between a R2R3 MYB and a basic helix-loop-helix MYC transcription factor related to flavonoid biosynthetic pathway in tobacco protoplasts.

Original languageEnglish
Pages (from-to)185-93
Number of pages9
JournalMethods in Molecular Biology
StatePublished - 2011


  • Arabidopsis Proteins/genetics
  • Fluorescent Dyes/analysis
  • Gene Expression Regulation, Plant/genetics
  • Plant Proteins/genetics
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Interaction Domains and Motifs/genetics
  • Protein Interaction Mapping/methods
  • Proto-Oncogene Proteins c-myc
  • Protoplasts/cytology
  • Tobacco/genetics
  • Transcription Factors/genetics


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