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 analyzing RNA-protein interactions in vivo. A major drawback of the use of such a transcriptional reporter-based assay in a library screen is the frequent occurrence of false-positive results due to bait RNA-independent activat...
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| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
1999-06-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/99266st01 |
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| Summary: | The yeast three-hybrid system presents a valuable tool for detecting and analyzing RNA-protein interactions in vivo. A major drawback of the use of such a transcriptional reporter-based assay in a library screen is the frequent occurrence of false-positive results due to bait RNA-independent activation of the reporter gene. To minimize the isolation of false positives in three-hybrid library screens, we incorporated a rapid and simple procedure based on differential sensitivity to 5-fluro-orotic acid. The technique effectively eliminates bait RNA-independent false positives and thus greatly enhances the efficiency of the yeast three hybrid system. |
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| ISSN: | 0736-6205 1940-9818 |