Abstract: The presently disclosed subject matter provides high-throughput assays, e.g., yeast three-hybrid assays, for detecting the presence of a molecule, e.g., therapeutic molecule. In certain embodiments, the presently disclosed subject matter provides genetically-engineered cells and assays for detecting tetracycline or derivatives thereof and kits for performing such assays.

Abstract: The disclosed subject matter relates to nucleic acid constructs that encode one or more fluorescent proteins, which can function as intracellular fluorescent tags, specifically for flow cytometry and fluorescence microscopy. The present disclosure further provides genetically-engineered cells that include one or more nucleic acid constructs and methods for using such genetically-engineered cells and nucleic acid constructs.

Abstract: The present disclosure provides a high-throughput fluorescence polarization microorganism screening assay for a metabolite produced by a microorganism. The metabolite is detected using competition with a fluorescent reporter for a receptor that binds both the metabolite and the reporter. The present disclosure further provides materials used in such an assay, a kit for such an assay, an isolated microorganism identified using such an assay, and culture conditions identified using such an assay.

Abstract: The present disclosure provides a high-throughput fluorescence polarization microorganism screening assay for a metabolite produced by a microorganism. The metabolite is detected using competition with a fluorescent reporter for a receptor that binds both the metabolite and the reporter. The present disclosure further provides materials used in such an assay, a kit for such an assay, an isolated microorganism identified using such an assay, and culture conditions identified using such an assay.

Abstract: The present invention relates to systems which harness the molecular biology of a living cell to direct evolution of biological entities of interest. According to the invention, a host cell is engineered to facilitate mutation of a nucleic acid target corresponding to that entity and select for desirable mutants. As applied to populations of host cells, the invention provides a means to generate and contemporaneously select mutants of interest, allowing for the production of extremely diverse libraries enriched in the most ‘fit’ mutants.

Abstract: A method for identifying a molecule that binds a known target in a cell from a pool of candidate molecules, comprising: (a) covalently bonding each molecule in the pool of candidate molecules to a methotrexate moiety to form a screening molecule; (b) introducing the screening molecule into a cell which expresses a first fusion-protein comprising a binding domain capable of binding methotrexate, a second fusion protein comprising the known target, and a reporter gene wherein expression of the reporter gene is conditioned on the proximity of the first fusion protein to the second fusion protein; (c) permitting the screening molecule to bind to the first fusion protein and to the second fusion protein so as to activate the expression of the reporter gene; (d) selecting which cell expresses the reporter gene; and (e) identifying the small molecule that binds the known target.

Abstract: The present invention relates to systems which harness the molecular biology of a living cell to direct evolution of biological entities of interest. According to the invention, a host cell is engineered to facilitate mutation of a nucleic acid target corresponding to that entity and select for desirable mutants. As applied to populations of host cells, the invention provides a means to generate and contemporaneously select mutants of interest, allowing for the production of extremely diverse libraries enriched in the most ‘fit’ mutants.

Abstract: A method for identifying a molecule as being able to bind a protein target in a cell, comprising (a) covalently bonding the molecule to trimethoprim (TMP) to form a screening molecule, (b) introducing the screening molecule into the cell which (A) expresses (i) a first fusion protein comprising a binding domain capable of binding TMP, and (ii) a second fusion protein comprising the protein target, and (B) comprises a reporter gene, wherein one of the fusion proteins comprises a DNA-binding domain and the other fusion protein comprises a transcription activation domain and wherein expression of the reporter gene is conditioned on the proximity of the DNA-binding domain to the transcription activation domain; and (c) determining whether the screening molecule binds to the first fusion protein and to the second fusion protein by determining whether the cell expresses the reporter gene, wherein expression of the reporter gene indicates that the molecule binds the protein target and wherein lack of expression of t

Abstract: A method for identifying a protein as being able to bind a ligand comprising, providing a molecule composed of a methotrexate moiety that is covalently bonded to the ligand; introducing the molecule into a cell which a) expresses a first fusion protein comprising a dihydrofolate reductase capable of binding methotrexate, expresses b) a second fusion protein comprising the protein, wherein one of the first and second fusion proteins also comprises a transcription activator domain and the other comprises a DNA-binding domain, and c) has a reporter gene wherein expression of the reporter gene is conditioned on the proximity of the first fusion protein to the second fusion protein; permitting the molecule to bind to the first fusion protein and to the second fusion protein so as to activate the expression of the reporter gene; and selecting the cell if it expresses the reporter gene.

Abstract: A transgenic bacterial cell comprising (a) a dimeric small molecule which comprises a first moiety known to bind a first receptor domain covalently linked to a second moiety known to bind a second receptor domain; (b) nucleotide sequences which upon transcription encode i) a first fusion protein comprising the first receptor domain, and ii) a second fusion protein comprising the second receptor domain; and (c) a reporter gene wherein expression of the reporter gene is conditioned on the proximity of the first fusion protein to the second fusion protein. The cell is also adapted for use in a method for identifying a molecule that binds to a known target in a bacterial cell from a pool of candidate molecules, and a method for identifying an unknown target receptor to which a molecule is capable of binding in a bacterial cell. Also described are compounds and kits for carrying out the methods.

Abstract: The subject invention provides a compound having the formula:

Abstract: A transgenic bacterial cell comprising

Abstract: A method for screening a cDNA library by identifying the expressed protein target, comprising:

Abstract: Described are compounds having the formula:

Abstract: The subject invention provides a compound having the formula: