Invitation to accurate BiFC assay : imaging of protein-protein interaction using fluorescent protein

Abstract

Protein-protein interaction plays a fundamental role to maintain biological process. Bimolecular fluorescence complementation （BiFC） assay is an imaging technique to visualize protein-protein interaction in living cells. BiFC assay is based on structural complementation between two non-fluorescent N-terminal and C-terminal fragments derived from a fluorescent protein. Over the past decade, BiFC assay has been widely used in plant science fields, due to its technical simplicity. However, designing appropriate control experiment is quite important for BiFC assay, because non-specific self-assembly of the non-fluorescent fragments induces background fluorescence, which may lead to misinterpretation of BiFC results. In this technical note, we describe information regarding fluorescent protein and BiFC assay in plants, and introduce “BiFC competition assay” as a control experiment.