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

Sitakanta Pattanaik, Joshua R. Werkman, Ling Yuan

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

6 Scopus citations

Abstract

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
Title of host publicationPlant Transcription Factors
Subtitle of host publicationMethods and Protocols
EditorsLing Yuan, Sharyn Perry
Pages185-193
Number of pages9
DOIs
StatePublished - 2011

Publication series

NameMethods in Molecular Biology
Volume754
ISSN (Print)1064-3745

Keywords

  • Bimolecular fluorescent complementation
  • protein-protein interaction
  • protoplast expression
  • split YFP
  • transcription factors

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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