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: 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: Reagents and compositions for use in reactions catalysed by luciferase enzymes, and in particular for use in luciferase-based gene reporter assays are described. The invention also provides methods and compositions for, inter alia, increasing the sensitivity and/or improving the kinetics of luciferase-catalysed reactions.

Abstract: A luminescence method for a luciferin/luciferase system which comprises reacting a luciferin solution containing an increased amount of dissolved carbon dioxide with luciferase; and a luciferase assay reagent comprising a luciferin solution containing an increased amount of dissolved carbon dioxide. Thus, a method of luminescent assay excellent in continuity and stability is provided as a substitute for the conventional luminescent luciferase assay featured by rapidity and high sensitivity.

Abstract: A codon optimized and stabilized luciferase gene based upon the sequence of the natural luciferase gene isolated from Luciola cruciata (Japanese firefly) and a novel recombinant DNA characterized by incorporating this new gene coding for a novel luciferase into a vector DNA for improved activities in mammalian cells, are disclosed. This new luciferase exhibits long-wavelength light emission, as well as improved thermostability and higher expression levels in mammalian cell systems, compared to native luciferase. Assays using this new enzyme for measuring various biological metabolic functions are described.

Abstract: A method and kit is provided for enhancing the tolerance of an assay reagent to compounds in an assay sample, the assay reagent including a luciferase enzyme. The method includes contacting the luciferase with a tolerance enhancement agent in an amount sufficient to substantially protect luciferase enzyme activity from interference of the compound and minimize interference by at least about 10% relative to an assay not having tolerance enhancement agent.

Abstract: Luciferase is conjugated to a chemical entity, particularly to a specific binding agent such as an antibody, antigen or a nucleic acid, and more particularly an antibody, by (a) mixing the luciferase with one or more of D-luciferin, magnesium ions and adenosine triphosphate and (b) performing a covalent coupling reaction between the luciferase and the binding reagent using a covalent coupling reagent where the amount of D-luciferin, magnesium ions and/or adenosine triphosphate is sufficient to protect the luciferase activity against inhibition by the covalent coupling reagent. Preferably, step (a) is carried out by mixing the luciferase with its substrates in solution and preferably both magnesium and adenosine triphosphate are present as magnesium adenosine triphosphate (Mg.sup.2+ ATP), optionally together with D-luciferin. Also disclosed is a labeled chemical entity comprising a chemical entity conjugated to active luciferase as formed by the method.

Abstract: Immunoassays which utilize an enzyme linked ligand or receptor wherein the enzyme is bacterial luciferase; mercantile kit useful in performing said immunoassay; and compounds utilized in performing said assay.

Abstract: The present invention provides a method of increasing the duration of detectable photon emission of a luciferase-luciferin reaction. The method provides a luciferase-luciferin reaction in which photon emission can be detected for up to and including eight hours. A method of the present invention can also be used to detect the presence of luciferase in biological samples. The present invention also provides a composition used in detecting the presence of luciferase in biological samples.

Abstract: The present invention provides a method of increasing the duration of detectable photon emission of a luciferase-luciferin reaction. The method provides a luciferase-luciferin reaction in which photon emission can be detected for up to and including eight hours. A method of the present invention can also be used to detect the presence of luciferase in biological samples. The present, invention also provides a composition used in detecting the presence of luciferase in biological samples.

Abstract: A method for detecting or quantitating a mutagenic substance in a sample includes culturing a host microorganism transformed with a recombinant gene comprising an SOS gene and genes expressing luciferase activity and optionally genes expressing an enzyme which catalyzes the production of a substrate for luciferase, positioned downstream of the SOS gene, in a medium to which the sample is added; and measuring a luminescence generated by expression of the gene expressing luciferase activity. The method is sensitive, accurate and non-time consuming; and gene systems used for said method, i.e., a recombinant gene comprising an SOS gene expressed when a DNA is damaged and a gene expressing luciferase activity positioned downstream of the SOS gene, and a host microorganism transformed with said recombinant gene. Preferably the recombinant gene further comprises a gene expressing an enzyme which catalyses the production of a substrate for the luciferase in the down stream of the SOS gene.

Abstract: A method of measuring the enzymatic activity of a luciferase includes contacting a luminogenic protein, such as a luciferase, with a protected luminophore to form a composition; and detecting light produced from the composition. The protected luminophore provides increased stability and improved signal-to-background ratios relative to the corresponding unmodified coelenterazine.

Abstract: A process for rapidly determining the bacterial count level in whey or whey constituents by adding a bacteria lysing reagent and then a luciferin-luciferase reagent to a sample to be analyzed at a pH from 7.0 to 7.9 and then measuring the bioluminescence light emitted from the luciferase-luciferin reaction, and an analysis kit for carrying out the process.

Abstract: A test kit and a rapid and sensitive method are provided for the multiple detection of organophosphate and carbamate pesticides. The test method employs an insect brain material that hydrolyzes a 6-substituted D luciferin ester, preferably a 6-substituted acetyl D luciferin ester, to produce D luciferin. Hydrolysis is inhibited by the pesticides in a test sample. Luciferase and adenosine triphosphate react with the D luciferin to produce oxyluciferin and emitted bioluminescence. The brain material is initially incubated with a test sample followed by the addition of the ester, luciferase and adenosine triphosphate. The kit contains the brain material, the ester and optionally luciferase and adenosine triphosphate, and may also contain means for incubating, and means for measuring emitted bioluminescence, and a control standard.

Abstract: Method for the bioluminescent dosing of enzymes, substrates or enzymatic inhibitors, characterized in that there is used a transparent support made of synthetic plastic material previously treated with a charged hydrophilic substance for the Fixation by adsorption of an enzymatic system containing at least a luciferase. The invention extends to a support of the polystyrene type treated by a charged hydrophilic substance on which an enzymatic system containing at least a luciferase is fixed by adsorption.

Abstract: Provided is a method of detecting infection in a wound caused by an infecting organism at a wound site. Also provided is a system for detecting an infection in a wound at a wound site. Additionally, a porous pad comprising luciferase is provided.

Abstract: An improved method for determining a viable microbial cell count in a sample wherein the method comprises the steps of filtering a microbe-containing sample solution through a hydrophilic membrane, thereby trapping viable microbes thereon, then spraying an extracting reagent onto the viable microbes trapped on the membrane, thereby lysing the microbial cells. The lysed cells are then sprayed with a solution of luciferin-luciferase reagent, thereby inducing a luminesent signal. The signal is then quantified using a CCD-based device. The improvement comprises the use of a volatile alcohol extracting reagent for lysing the viable microbes, then evaporating the alcohol prior to spraying the luciferin-luciferase solution, leaving a concentrated ATP,containing solution for enhanced luminescent detection.

Abstract: The number of bacteria in a cell population comprising bacteria and somatic cells is determined by concentrating the bacteria and the somatic cells on the surface of a substrate of a filter material, such as a membrane filter, rupturing the somatic cells by treatment with a medium which is not in osmotic balance with the medium inside the cells, such as water which here constitutes a hypotonic medium, and removing or inactivating their ATP, establishing an extraction chamber having an inner wall which is partly defined by the bacteria-bearing surface of the filter material and therein extracting the ATP from the bacteria with a medium containing a surfactant, such as a quaternary ammonium compound, capable of perforating the cell walls of the bacteria, adding luciferin and luciferase and transferring the medium to a measuring chamber, determining the amount of ATP by measuring the light emitted as a result of the luciferin/luciferase reaction and expressing the amount of ATP as the number of bacteria present

Abstract: Reagents and compositions for use in reactions catalysed by luciferase enzymes, and in particular for use in luciferase-based gene reporter assays are described. The invention also provides methods and compositions for, inter alia, increasing the sensitivity and/or improving the kinetics of luciferase-catalysed reactions.