Grow'n'Glow GFP Yeast One-Hybrid System

The Grow'n'Glow GFP One-Hybrid kit isolates genes for proteins that bind a specific DNA element of interest. This one-hybrid system can be used to investigate the bases and amino acids involved in specific DNA-protein interactions. The Grow'n'Glow system offers maximum sensitivity because detection of the DNA-protein interaction occurs in vivo, where proteins are more likely to be in their native conformation.

Use of the Green Fluorescent Protein (GFP) reporter eliminates the need for time-consuming beta-galactosidase lift assays. The GFP variant expressed by the GFP reporter plasmid, GFPuv, has the same excitation and emission maxima as wild-type GFP, but is 18 times brighter than the wild-type variant. Positive colonies with DNA-binding proteins bright glow green. The glowing colonies are easily detected by placing the plate with the yeast colonies on a standard 300 nm UV transilluminator or under a UV lamp in a darkroom.

Expression of fusion proteins by the prey plasmid is controlled by the GAL1 upstream-activation sequence. In yeast with an intact galactose regulatory system, the GAL1 activation sequence is induced by galactose and repressed by glucose. This regulation inhibits expression of library/activation domain fusions until the actual screening, which prevents any potentially toxic library proteins from affecting growth of the yeast. These toxic proteins would be missed during screening in a non-inducible system.