METHOD FOR SCREENING OF PROTEIN-PROTEIN INTERACTION BY DOUBLE LIBRARY TWO-HYBRID SYSTEM AND RECOMBINANT YEAST STRAINS USED THEREFOR

摘要

PURPOSE: A method for screening of protein-protein interaction by double library two-hybrid system and recombinant yeast strains used therefor are provided, thereby simultaneously and effectively screening the coupling pairs of library proteins. CONSTITUTION: A method for screening of protein-protein interaction by double library two-hybrid system comprises the steps of: (a) preparing a type yeast strain in which URA3 is destroyed, and inserting a plasmid in which URA3 is expressed by a promoter GAL1 into the a type yeast strain; (b) directly correcting the yeast strain in order for URA3 within the yeast strain to be expressed by a promoter GAL1 using a gene recombination method to prepare a type alpha yeast strain; (c) transforming the a type yeast strain and an alpha type yeast strain with bait library and prey library; (d) independently culturing the transformed a type yeast strain and the alpha type yeast strain in a medium containing 5-fluoroorotic acid(5-FOA) to selectively remove the false positive reaction; and (e) mating two types of transformed yeast strains to detect protein-protein interaction pairs.