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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| Format: | Article |
| Language: | English |
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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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| _version_ | 1850151672941117440 |
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| author | Young Woo Park Seng-Lai Tan Michael G. Katze |
| author_facet | Young Woo Park Seng-Lai Tan Michael G. Katze |
| author_sort | Young Woo Park |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-fb21f8fd688547c9b113a7424efbd1ce |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 1999-06-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-fb21f8fd688547c9b113a7424efbd1ce2025-08-20T02:26:09ZengTaylor & Francis GroupBioTechniques0736-62051940-98181999-06-012661102110610.2144/99266st01Differential Sensitivity to 5-fluoro-orotic Acid as a Screen for Bait RNA Independent False Positives in a Yeast Three-Hybrid SystemYoung Woo Park0Seng-Lai Tan1Michael G. Katze21University of Washington, Seattle, WA, USA1University of Washington, Seattle, WA, USA1University of Washington, Seattle, WA, USAThe 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.https://www.future-science.com/doi/10.2144/99266st01 |
| spellingShingle | Young Woo Park Seng-Lai Tan Michael G. Katze Differential Sensitivity to 5-fluoro-orotic Acid as a Screen for Bait RNA Independent False Positives in a Yeast Three-Hybrid System BioTechniques |
| title | Differential Sensitivity to 5-fluoro-orotic Acid as a Screen for Bait RNA Independent False Positives in a Yeast Three-Hybrid System |
| title_full | Differential Sensitivity to 5-fluoro-orotic Acid as a Screen for Bait RNA Independent False Positives in a Yeast Three-Hybrid System |
| title_fullStr | Differential Sensitivity to 5-fluoro-orotic Acid as a Screen for Bait RNA Independent False Positives in a Yeast Three-Hybrid System |
| title_full_unstemmed | Differential Sensitivity to 5-fluoro-orotic Acid as a Screen for Bait RNA Independent False Positives in a Yeast Three-Hybrid System |
| title_short | Differential Sensitivity to 5-fluoro-orotic Acid as a Screen for Bait RNA Independent False Positives in a Yeast Three-Hybrid System |
| title_sort | differential sensitivity to 5 fluoro orotic acid as a screen for bait rna independent false positives in a yeast three hybrid system |
| url | https://www.future-science.com/doi/10.2144/99266st01 |
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