Protein-protein interaction is a key concept in the fields of signal transduction and gene regulation. Investigators can make educated guesses as to whether two proteins actually interact, but verifying these interactions is not a trivial task. Classical biochemical techniques are fraught with pitfalls that make data acquisition and interpretation difficult. BioSignal Packard, a division of Packard Instruments, has introduced the BRET2 (Bioluminescence Resonance Energy Transfer) system to assess these protein-protein interactions in vivo.
Components of the BRET2 system This technology uses Renilla luciferase (Rluc) as the donor and a modified Green Fluorescent Protein (GFP2) as the acceptor molecule in an assay analogous to fluorescence resonance energy transfer (FRET), but without the need for, or problems associated with, an excitation light source. The system consists of two eukaryotic expression constructs and a proprietary Rluc substrate. The two proteins of interest are cloned into vectors to produce fusions with Rluc and GFP2. ...
