Biological expression reporter gene

The yeast two-hybrid system is a powerful genetic technique that has been used successfully to study protein-protein interactions in vivo from a wide variety of biological sources, including yeast, plant, and mammalian (Fields and Song, 1989). The two-hybrid system provides a sensitive method to detect relatively weak and transient protein-protein interactions that may not be biochemically detectable. The principle of this assay is based on the reconstitution of transcriptional factors that promotes the expression reporter genes, which then allow the proliferation of yeast under restrictive conditions. Using the two-hybrid system, we successfully isolated IPCEFl and a novel kinesin motor proteins from a rat brain cDNA library as interactors of cytohesin 2 ARF GEF and centaurin-al ARF GAP, respectively (Venkateswarlu, 2003). The procedure used to map the binding domains of cytohesin 2 and IPCEFl, and to characterize the cytohesin 2 and IPCEFl selectivity is outlined here. 

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