Screening for in planta protein-protein interactions combining bimolecular fluorescence complementation with flow cytometry.

ABSTRACT: Understanding protein and gene function requires identifying interaction partners using biochemical, molecular or genetic tools. In plants, searching for novel protein-protein interactions is limited to protein purification assays, heterologous in vivo systems such as the yeast-two-hybrid or mutant screens. Ideally one would be able to search for novel protein partners in living plant cells. We demonstrate that it is possible to screen for novel protein-protein interactions from a random library in protoplasted Arabidopsis plant cells and recover some of the interacting partners. Our screen is based on capturing the bi-molecular complementation of mYFP between an YN-bait fusion partner and a completely random prey YC-cDNA library with FACS. The candidate interactions were confirmed using i

SUBMITTER: Berendzen KW 

PROVIDER: S-EPMC3458939 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature