Rapid selection against truncation mutants in yeast reverse two-hybrid screens
- Author(s)
- Puthalakath, H; Strasser, A; Huang, DCS;
- Details
- Publication Year 2001-05-01,Volume 30,Issue #5,Page 984-+
- Journal Title
- BIOTECHNIQUES
- Publication Type
- Journal Article
- Abstract
- 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.
- Publisher
- EATON PUBLISHING CO
- Keywords
- PROTEIN-PROTEIN INTERACTIONS; SYSTEMS
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2001-05-01 12:00:00