Ljubljana/splittevpassay

Split TEV protease

TEV protease can be split into two stable domains(N-terminal and C-terminal). These halves are inactive but can assemble and regain activity when they are brought together after dimerization of transmembrane proteins. For detecting TEV protease activity, another fusion protein must be constructed. It is composed of a membrane anchor, linker, containing a specific TEVP cleavage site and a transcription factor. When both halves of TEV protease assemble, split protease regains activity, cleaves off the substrate protein, thus releasing the transcription factor from the membrane. Further, the transcription factor is translocated to the nucleus where it induces transcription of a reporter gene.