bacterial two hybrid based on λcI

It may be part of transcriptional complementation assay (MI:0232). The authors often refer it as 'bacterial two hybrid' but many are called two-hybrid (even in E. Coli) and it may be a bit confusing for curation. This one really looks like two hybrid CV as there is a DNA binding domain however the other fusion is directly a RNA polymerase so it might need another specific CV for it. The BacterioMatch II two-hybrid is name of a kit sold by stratagen so it might would need another name. 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 λ repressor protein (λcI, 237 amino acids), containing the amino-terminal DNA-binding domain and the carboxylterminal dimerization domain. The corresponding target protein is fused to the N-terminal domain of the α-subunit of RNA polymerase (248 amino acids). The bait is tethered to the λ operator sequence upstream of the reporter promoter through the DNA-binding domain of λcI. 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 (Figure 1). Due to the tendency of both the λ repressor protein and the N-terminal domain of the α-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.

used in : PMID 15792953 http://www.stratagene.com/manuals/200414.pdf

Reported by: krobbe4

HUPO-PSI / psi-mi-CV

bacterial two hybrid based on λcI #53