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.

Abstract: The presence of renilla luciferase alone or both renilla luciferase and firefly luciferase is detected by adding reagent mixture(s) to a biological sample and producing glow luminescence having a duration of at least an hour. In producing luminescence from renilla luciferase alone, a reagent is added comprising coelenterazine, and dithiothreitol (DTT) and EDTA, or functional equivalents of DTT and EDTA. Luminescence from samples containing both firefly luciferase and renilla luciferase is produced by first adding a reagent comprising firefly luciferin, ATP, co-factors necessary for firefly luciferase activity (e.g., Ca+2 and Mg+2), dithiothreitol (DTT) or functional equivalents thereof, and AMP. Following measurement of the firefly luciferase, coelenterazine and EDTA, or functional equivalents of EDTA, are added, and the luminescence produced by renilla luciferase is measured.

Abstract: The present invention is one gene construct or a combination of two gene constructs or expression vectors incorporating a Cypridina luciferase gene and a copepod luciferase under the control of distinct promoters. These gene constructs and expression vectors are useful for making a mammalian cell incorporating the Cypridina luciferase gene and the copepod luciferase to be capable of stably expressed and extracellularly secreted under the control of the distinct promoters.

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: The present invention is one gene construct or a combination of two gene constructs or expression vectors incorporating a Cypridina luciferase gene and a copepod luciferase under the control of distinct promoters. These gene constructs and expression vectors are useful for making a mammalian cell incorporating the Cypridina luciferase gene and the copepod luciferase to be capable of stably expressed and extracellularly secreted under the control of the distinct promoters.

Abstract: The present invention relates to a method for producing Cypridina luciferase labeled with hydrophilic biotin, characterized in that a biotin reagent containing a polyalkylene glycol structure as a spacer is reacted with Cypridina luciferase, and biotin-labeled Cypridina luciferase wherein a sugar chain in Cypridina luciferase has been biotinylated.

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 and kit are 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 minize interference by at least about 10% relative to an assay not having tolerance enhancement agent.

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: Plasmids and methods of use for assaying translational recoding are disclosed. The plasmids contain a constitutively expressed renilla (Renilla reniformis; sea pansy) luciferase gene, a polylinker for insertion of a selected DNA segment, and an out-of-frame firefly luciferase gene. Recoding is determined by monitoring luminescence of the firefly luciferase normalized to the luminescence of the renilla luciferase.

Abstract: Inhibitors of luciferase enzymes are disclosed and find use in multiplexed assays using multiple luciferases and multiple inhibitors, in both in vitro and in vivo embodiments.

Abstract: The present invention relates to a luciferase derived from a star-worm belonging to genus Diplocladon, the luciferase inducing luminescence such that a maximum luminous wavelength falls within a range of 557 to 562 nm over the entire pH range of 5.5 to 8.0, or the luciferase inducing luminescence having 1.5 times the luminous intensity of luminescence induced by Photinus pyralis firefly luciferase.

Abstract: According to one embodiment, the present invention relates to luciferase derived from Malaysian Luciola firefly, the luciferase having a maximum luminescent wavelength of 580 nm at pH 8, or the luciferase indicating 23.3 times or more of luminescent intensity in comparison to that of Rhodamine 6G.

Abstract: A method and kit are 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: 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: Disclosed is a purified luciferase and a method for making it. The luciferase is obtained from Luciola cruciata. The luciferase has a pH range for stabililty of 6.5-9.0 and a optimum pH range of 8.0-9.5. The enzyme does not act on ADP, CTP, UTP and GTP.

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 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 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: The present invention relates to biotinated firefly luciferase comprising a biotinated peptide and firefly luciferase linked therein, biotinated firefly luciferase having a specific amino acid sequence, a biotinated firefly luciferase gene comprising a gene coding for a biotinated peptide and a firefly luciferase gene linked therein, a biotinated firefly luciferase gene comprising a biotinated peptide gene coding for a specific amino acid sequence and a firefly luciferase gene linked therein, a recombinant DNA comprising said biotinated firefly luciferase gene inserted into a vector DNA, a process for producing biotinated firefly luciferase comprising culturing a microorganism belonging to the genus Escherichia carrying said recombinant DNA and then recovering the resulting biotinated firefly luciferase from the culture, a bioluminescent analysis method comprising using said biotinated firefly luciferase, and a bioluminescent analysis method comprising quantifying a ligand by measuring a biotinated receptor b