Rapid selection against truncation mutants in yeast reverse two-hybrid screens
H Puthalakath, A Strasser, DCS Huang
BIOTECHNIQUES | EATON PUBLISHING CO | Published : 2001
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.
Awarded by NCI NIH HHS
Awarded by NATIONAL CANCER INSTITUTE