Rapid selection against truncation mutants in yeast reverse two-hybrid screens
The yeast reverse two-hybrid system is a powerful technique for isolating mutations in a protein that abolish its interaction with a known partner. Selection is based on abrogation of growth suppression imposed when wild-type interactions confer 5-fluoroorotic acid (5-FOA) sensitivity to yeast cells...
| Veröffentlicht in: | BioTechniques. - 1993. - 30(2001), 5 vom: 01. Mai, Seite 984-8 |
|---|---|
| 1. Verfasser: | |
| Weitere Verfasser: | , |
| Format: | Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2001
|
| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Technical Report Journal Article Apoptosis Regulatory Proteins Bcl-2-Like Protein 11 Carrier Proteins Drosophila Proteins Luminescent Proteins Membrane Proteins mehr... |
| Zusammenfassung: | The yeast reverse two-hybrid system is a powerful technique for isolating mutations in a protein that abolish its interaction with a known partner. Selection is based on abrogation of growth suppression imposed when wild-type interactions confer 5-fluoroorotic acid (5-FOA) sensitivity to yeast cells. A laborious component of this system is to eliminate those mutations that cause protein truncation. By fusing the green fluorescent protein (GFP) to the C-terminus of a protein of interest, dynein light chain (LC8), we were able to rapidly isolate mutations that did not result in protein truncation |
|---|---|
| Beschreibung: | Date Completed 04.12.2001 Date Revised 28.09.2018 published: Print ErratumIn: Biotechniques 2001 Sep;31(3):488 Citation Status MEDLINE |
| ISSN: | 0736-6205 |