Periplasmic membrane-bound system for detecting...

Details Periplasmic membrane-bound system for detecting... Periplasmic membrane-bound system for detecting...

C - Chemistry, Metallurgy

12

N

C12N 15/62 (2006.01) C07K 14/28 (2006.01) C07K 19/00 (2006.01) C12N 1/21 (2006.01) C12N 15/31 (2006.01) C12N 15/63 (2006.01) C12Q 1/68 (2006.01) G01N 33/53 (2006.01) C07K 14/035 (2006.01) C07K 14/16 (2006.01)

Patent

CA 2117612

The present invention relates to: (1) a fusion protein having a dimerizing domain with or without a ligand-binding region and tox R DNA-binding and hydrophobic transmembrane regions; (2) host cells comprising the fusion protein and a nucleic acid molecule having a reporter gene operatively linked to the ctx operon, wherein dimerization (ligand-dependent or -independent) is signaled by expression of the reporter gene; (3) a nucleic acid molecule coding for the fusion protein; (4) an expression vector comprising a coding region for the fusion protein; (5) a process for detecting dimer formation (ligand dependent or ligand independent) of the fusion protein, which comprises treating a culture of the host cells with a ligand, ligand mimetic, or dimerization inhibitor, and screening for expression of the reporter gene. The present invention can be used to generate a signal from a variety of ligand-binding domains, allowing ligand binding to be indicated by a simple colorimeteric test or antibiotic resistance. The fusion proteins could include therapeutically relevant domains, so that biologically pertinent interactions can be indicated by a readily measurable signal.

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