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US 2011/0237451 A1Published application
Competitive n-hybrid system
AssigneeSignalomics GmbH
InventorsClaudia Arntz, Danielle Meinders, Christoph Block
Life of this patent
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AssigneeSignalomics GmbH
InventorsClaudia Arntz, Danielle Meinders, Christoph Block
Estimating the term…
A method of identifying high-affinity ligands, comprising the following steps: a) generating a library for a mutagenized first hybrid protein, comprising a multiplicity of mutants; b) expressing said first hybrid protein in a host with a second hybrid protein, with one of said hybrid proteins comprising the DNA binding domain of a transcription factor and a bait protein, and the other hybrid protein comprising the activating domain for a transcription factor and a prey protein; c) enabling said first and second hybrid proteins to bind to one another to give a complex containing a functional transcription factor in the host cell under reaction conditions chosen so as to shift the equilibrium of the binding reaction toward the side of the hybrid proteins; d) detecting the binding reaction by detecting a reporter gene expressed via the functional transcription factor; e) optionally repeating one or more steps from a) to d); f) selecting a mutant.
A method of identifying high-affinity ligands, comprising the following steps:
A host cell coding for a first hybrid protein and a second hybrid protein, it being possible for said hybrid proteins to form together a functional ligand complex, and a protein which is a competitor of either of said hybrid proteins.
A system of a host cell coding for a first hybrid protein and a second hybrid protein, it being possible for said hybrid proteins to form together a functional ligand complex, and of a protein which is a competitor of either of said hybrid proteins.
A plasmid coding for a first hybrid protein which, together with a second hybrid protein, forms a ligand complex, and for another protein which is a competitor of said first or second hybrid protein.
The use of a fluorophore whose maximum emission is between 550 and 700 nm, preferably between 580 and 650, particularly preferably between 600 and 620, especially preferably between 600 and 610 nm as readout in yeasts in which at least two hybrid proteins are coexpressed.
A fluorophore encoded by any of the following sequences:
Dependencies are inferred from the claim text.