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.
|Title of host publication||Plant Transcription Factors|
|Subtitle of host publication||Methods and Protocols|
|Number of pages||9|
|State||Published - 2011|
|Name||Methods in Molecular Biology|
Bibliographical noteFunding Information:
We thank Dr. E. Grotewold of the Ohio State University for the split YFP vectors. We also express our appreciation to Dr. S. Wessler of the University of Georgia for the Lc cDNA. This work is supported by a grant from the Kentucky Tobacco Research and Development Center to L.Y.
This work is supported by a grant from the Kentucky Tobacco Research and Development Center to L.Y.
© 2011, Springer Science+Business Media, LLC.
- Bimolecular fluorescent complementation
- protein–protein interaction
- protoplast expression
- split YFP
- transcription factors
ASJC Scopus subject areas
- Molecular Biology