SYSTEM FOR SCREENING PROTEIN OR PEPTIDE LOCALIZED IN SPECIFIC AREA IN CELL

摘要

By integrating a tetracycline repressor (TetR)/ operator (TetOP) system into a yeast, detection and screening are conducted with the use of a change in the intracellular localization of a tetracycline repressor-fused protein as an indication. A TetR-fused protein (11) expressed from a TetR expression plasmid library migrates into a nucleus (14) under the action of a nuclear transport signal. The TetR-fused protein (11) binds to TetOP (12) inserted into a HIS3 reporter gene promoter and thus regulates the HIS3 reporter gene (13). In the case where the protein having been fused with TetR has an activity of controlling the transport toward a specific area (15) in a cell, The TetR-fused protein (11) is discharged from the nucleoplasm and thus the expression of the HIS3 reporter gene (13) is induced. Accordingly, an activity of transporting toward a specific area in a cell can be detected by selecting a clone capable of growing a minimum medium deficient in histidine.