Abstract: Processes are disclosed using the depolymerization of a nucleic acid hybrid to qualitatively and quantitatively analyze for the presence of a predetermined endogenous nucleic acid. Applications of those processes include the detection of single nucleotide polymorphisms, identification of single base changes, speciation, determination of viral load, genotyping, medical marker diagnostics, and the like.

Abstract: The invention provides active, non-naturally occurring mutants of beetle luciferases and DNAs which encode such mutants. A mutant luciferase of the invention differs from the corresponding wild-type luciferase by producing bioluminescence with a wavelength of peak intensity that differs by at least 1 nm from the wavelength of peak intensity of the bioluminescence produced by the wild-type enzyme. The mutant luciferases and DNAs of the invention are employed in various biosensing applications.

Abstract: Processes are disclosed using the depolymerization of a nucleic acid hybrid to qualitatively and quantitatively analyze for the presence of a predetermined nucleic acid. Applications of those processes include the detection of single nucleotide polymorphisms, identification of single base changes, speciation, determination of viral load, genotyping, medical marker diagnostics, and the like.

Abstract: Processes are disclosed using the depolymerization of a nucleic acid hybrid to qualitatively and quantitatively analyze for the presence of predetermined nucleic acid target sequences using a multiplex assay format. Applications of those processes include the detection of single nucleotide polymorphisms, identification of single base changes, speciation, genotyping, medical marker diagnostics, and the like.

Abstract: Processes are disclosed using the depolymerization of a nucleic acid hybrid to qualitatively and quantitatively analyze for the presence of a predetermined exogenous nucleic acid. Applications of those processes include the detection of single nucleotide polymorphisms, identification of single base changes, determination of viral load, genotyping, medical marker diagnostics, and the like.

Abstract: Processes are disclosed using the depolymerization of a nucleic acid hybrid to qualitatively and quantitatively analyze for the presence of a predetermined nucleic acid. Applications of those processes include the detection of single nucleotide polymorphisms, identification of single base changes, speciation, determination of viral load, genotyping, medical marker diagnostics, and the like.

Abstract: Methods and compositions are provided for improved kinetics of light production from luciferase activity in beetle luciferase-luciferin reactions utilizing a luciferase inhibitor. Thus, the invention provides methods and compositions for assaying samples for the presence of a beetle luciferase in conjunction with a luciferase inhibitor.

Abstract: The presently disclosed invention is drawn to an automatic luminometer apparatus capable of measuring two distinct luminescent reactions from within a single, non-compartmentalized sample container. The present apparatus may be dimensioned, configured, and programmed to automatically perform dual-reporter luminescent assays using multi-well sample plates, such as 96-well microtiter plates.

Abstract: The present invention relates to single and dual-reporter luminescence assays utilizing general and specific reagents to quench enzyme-mediated reactions. In one embodiment of the invention, a reagent is added to the assay which non-specifically quenches enzyme-mediated luminescent reactions. In another embodiment of the invention, a reagent is added to the assay which simultaneously quenches one enzyme-mediated luminescent reaction while activating another distinct enzyme-mediated luminescent reaction. An assay kit containing specific quench reagents, and the reagents themselves are also disclosed.

Abstract: A method is disclosed for producing a protein which expresses bioluminescence activity which involves combining two polydeoxyribonucleotides, one containing a continuous sequence of codons encoding a polypeptide which comprises a single covalently bonded molecular structure and which catalyzes the oxidation of insect luciferin to yield light and the other which causes DNA transcription, and obtaining the polypeptide by transcription and subsequent translation. The insect luciferin is derived from bioluminescent insect, preferably Diptera and Coleoptera (fireflies and beetles). Hybrid proteins are similarly formed by inclusion of an additional polydeoxyribonucleotide encoding for a second polypeptide such that their respective polypeptide-encoding reading frames form a continuous reading frame.

Abstract: A modified form of beetle luciferase, which has been engineered for improved genetic reporting, is disclosed. The modified form contains one or more new features. Chief among these is removal of the peroxisomal translocation sequence to yield a cytoplasmic form of the enzyme. Other changes include removal of potentially interfering restriction sites and genetic regulatory sites from the gene, improvement of the codon usage for mammalian cells. The modified luciferase reporter enzyme is also devoid of potential N-glycosylation targets to minimize post-translational modification and remains in the cytoplasm of host cells to optimize substrate availability.

Abstract: Methods and compositions are provided for improved kinetics of light production from luciferase activity in beetle luciferase-luciferin reactions. Thus, the invention provides methods, compositions and test kits for assaying samples for the presence of a beetle luciferase or, using a beetle luciferase-luciferin reaction, the presence of ATP. The invention also provides the complex of the coenzyme, Coenzyme A, a beetle luciferase and oxyluciferin in its exited state in the luciferase-luciferin reaction and luciferyl-CoA, the thioester of Coenzyme A and luciferin (D-(-)-2-(6'-hydroxy-2'-benzothiazolyl)-.DELTA..sup.2 -thiazoline-4-carboxylic acid).

Abstract: Methods and compositions are provided for improved kinetics of light production from luciferase activity in beetle luciferase-luciferin reactions. Thus, the invention provides methods, compositions and test kits for assaying samples for the presence of a beetle luciferase or, using a beetle luciferase-luciferin reaction, the presence of ATP. The invention also provides the complex of the coenzyme, Coenzyme A, a beetle luciferase and oxyluciferin in its exited state in the luciferase-luciferin reaction and luciferyl-CoA, the thioester of Coenzyme A and luciferin (D-(-)-2-(6'-hydroxy -2'-benzothiazolyl)-.DELTA..sup.2 -thiazoline-4-carboxylic acid).

Abstract: A method is disclosed for producing a protein which expresses bioluminescence activity which involves combining two polydeoxyribonucleotides, one containing a continuous sequence of codons encoding a polypeptide which comprises a single covalently bonded molecular structure and which catalyzes the oxidation of insect luciferin to yield light and the other which causes DNA transcription, and obtaining the polypeptide by transcription and subsequent translation. The insect luciferin is derived from bioluminescent insect, preferably Diptera and Coleoptera (fireflies and beetles). Hybrid proteins are similarly formed by inclusion of an additional polydeoxyribonucleotide encoding for a second polypeptide such that their respective polypeptide-encoding reading frames form a continuous reading frame.

Abstract: The invention provides improved methods for assaying samples for the presence of a beetle luciferase. The methods of the invention entail improved kinetics of light production from luciferase activity in beetle luciferase-luciferin reactions.