BacterioMatch II Two-Hybrid System - Details & Specifications

The BacterioMatch II two-hybrid system detects protein-protein interactions based on transcriptional activation. A protein of interest (the bait) is fused to the full-length bacteriophage l repressor protein (lcI, 237 amino acids), containing the amino-terminal DNA-binding domain and the carboxyl-terminal dimerization domain. The corresponding target protein is fused to the N-terminal domain of the a-subunit of RNA polymerase (248 amino acids). The bait is tethered to the l operator sequence upstream of the reporter promoter through the DNA-binding domain of lcI.

When the bait and target interact, they recruit and stabilize the binding of RNA polymerase at the promoter and activate the transcription of the HIS3 reporter gene. A second reporter gene, aadA, encoding a protein that confers streptomycin resistance, provides an additional mechanism to validate the bait and target interaction.

Due to the tendency of both the  repressor protein and the N-terminal domain of the a-subunit of RNA polymerase to dimerize, this system might not be optimal for the analysis of proteins that self-associate unless their interaction with other protein partners depends on the oligomerization.

For Research Use Only. Not for use in diagnostic procedures