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Protein-fragment complementation assay, or PCA, is a method for the identification of protein–protein interactions in biological systems. It is used in the field of proteomics. In the PCA, the proteins of interest ("Bait" and "Prey") are each covalently linked to incomplete fragments of a third protein (e.g. DHFR, which acts as a "reporter"). Interaction between the "bait" and the "prey" proteins brings the fragments of the "reporter" protein in close enough proximity to allow them to form a functional reporter protein whose activity can be measured. This principle can be applied to many different "reporter" proteins and is also the basis for the yeast two-hybrid system, an archetypical PCA assay. ==Split protein assays== Any protein that can be split into two parts and reconstituted non-covalently may be used in a PCA. The two parts just have to be brought together by other interacting proteins fused to them (typically called "bait" and "prey" (see figure). The protein that produces a detectable readout is called "reporter". Usually enzymes which confer resistance to antibiotics, such as Dihydrofolate reductase or Beta-lactamase, or proteins that give colorimetric or fluorescent signals are used as reporters. When fluorescent proteins are reconstituted the PCA is called Bimolecular fluorescence complementation assay. The following proteins have been used in split protein PCAs: *Dihydrofolate reductase (DHFR) *Beta-lactamase *Yeast Gal4 (as in the classical yeast two-hybrid system) *Split TEV (Tobacco etch virus protease) *Luciferase,〔Fujikawa, Y. et al. (2014) (Split luciferase complementation assay to detect regulated protein-protein interactions in rice protoplasts in a large-scale format ). Rice 7:11〕 including ReBiL (recombinase enhanced bimolecular luciferase) *Ubiquitin *GFP (split-GFP), e.g. EGFP (enhanced green fluorescent protein) *LacZ (beta-galactosidase) * Infrared fluorescent protein IFP1.4, an engineered chromophore-binding domain (CBD) of a bacteriophytochrome from ''Deinococcus radiodurans'' * Focal adhesion kinase (FAK)〔Ma, Y. et al. (2014) (Split focal adhesion kinase for probing protein–protein interactions ). Biochemical Engineering Journal, 〕 抄文引用元・出典: フリー百科事典『 ウィキペディア（Wikipedia）』 ■ウィキペディアで「Protein-fragment complementation assay」の詳細全文を読む スポンサード リンク
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