REVERSE TWO-HYBRID METHOD USING GALACTOSE SENSIBILITY

摘要

PROBLEM TO BE SOLVED: To provide a reverse two-hybrid method capable of detecting interaction and/or non-interactive action between proteins by a simple system. SOLUTION: The detection of interaction and/or non-interactive action between proteins X and Y is conducted by adopting a transformant where a galactose sensibility microorganism is transformed with an expression vector. The vector includes a DNA encoding protein complexes DB-X and AD-Y which are prepared by the fusion of proteins X and Y with an UAS binding domain (DB) and a transcription-activating domain(AD) in the transcription factor GAL4 of Saccharomyces yeast respectively. The gal 7 gene encoding galactose-1- phosphoric acid uridylyl transferase in the downstream of the UAS has been broken in the galactose sensibility microorganism. When non-interactive action between X and Y is found, galactode-1-phosphoric acid is not stored in the microorganism and the microorganism doesn't propagate in a medium including galactose but can propagate in a medium not including galactose. Thus, the non-interactive action between X and Y can be detected.