The yeast reverse two-hybrid system is a powerful technique for isolating m
utations in a protein that abolish its interaction with a known partner. Se
lection 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 truncatio
n.