Overview of our system

Vector information

Protocols

Protoplast isolation

Gene transfection

Luciferase assay

Western blotting

Results

Transfect efficiency

Gene expression levels of dual expression vectors

Protein interaction

Acknowledgements

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Arabidopsis
Fig. Complemented Renilla luciferase activities in Arabidopsis protoplasts expressing histone H2A and H2B

PPI interaction

Assay
PPI signal
Split luciferase
18
BRET*
1.04-1.3
FRET**
1.14-1.27

*Subramanian et al. 2006 Nat Biotechnol , 19;769-772
**Cheng et al. 2003 Genes&Development 13;334-344, Churchman et al., 2006 Plant Cell, 18;3145-3157

Split luciferase can detect protein-protein interaction at 10-fold higher than FRET and BRET assays

Rice
Fig. Complemented Renilla luciferase activities in Rice protoplasts expressing histone proteins and SYP proteins

Rice results

Protein-protein interactions detected by this method were published in the follwoing papers.

 

 

Last Update 11/28/09