That Certain Glow

Grow 'n' Glow GFP Two Hybrid System

Many genetic and cellular processes rely on highly specific protein-protein interactions. Two-hybrid systems provide a way to systematically identify pairs of proteins (referred to as "bait" and "prey" proteins) that form stable contacts, and further, to identify which protein segments or domains are necessary to form the complex. MoBiTec's Grow 'n' Glow GFP Two-Hybrid System combines a relatively straightforward two-hybrid strategy with inducible expression, visual hybrid identification, and convenient immunoprecipitation of positives.

Developed by scientists at the Max Planck Institute, the Grow 'n' Glow system uses three vectors: pGNG1, the GFP reporter plasmid; pEG202, the bait plasmid; and pJG4-5, the prey plasmid. Screening a known protein for contacts with a library of unknown proteins first requires cloning the known protein into the bait plasmid to produce an in-frame fusion between the known protein coding sequence and pEG202's LexA DNA-binding domain. This is the...