Differential sensitivity to 5-fluoro-orotic acid as a screen for bait RNA-independent false positives in a yeast three-hybrid system

The yeast three-hybrid system presents a valuable tool for detecting and an alyzing RNA-protein interactions in vivo. A major drawback of the use of su ch a transcriptional reporter-based assay in a library screen is the freque nt occurrence of false-positive results due to bait RNA-independent activat ion of the reporter gene, To minimize the isolation of false positives in t hree-hybrid library screens, we incorporated a rapid and simple procedure b ased on differential sensitivity to 5-fluro-orotic acid. The technique effe ctively eliminates-bait RNA-independent false positives and thus greatly en hances the efficiency of the yeast three-hybrid system.