Abstract: Nucleotide and amino acid sequences of luciferase peptides that are encoded by genes within the genome of Arachnocampa (Diptera) are disclosed. Specifically provided are functional ATP-dependent luciferases that catalyze luminescence reactions with emission spectra within the blue portion of the spectrum. The present invention specifically provides isolated peptide and nucleic acid molecules, methods of identifying orthologs and active subsequences of the enzyme peptides, and methods of identifying modulators and substrates of the luciferase peptides. Methods of assays, including multiple reporter assays utilizing at least two ATP-dependent luciferases are provided.

Abstract: This invention relates to: the development of a mutant firefly luciferase in order to use dATP as a DNA polymerase substrate upon pyrosequencing, such luciferase being subjected to substrate specificity modification in a manner such that the dATP-induced activity alone is decreased while the ATP-induced activity is maintained; and a mutant firefly luciferase for which the proportion of activity induced by dATP to activity induced by ATP (dATP/ATP) is lower than that for the wild-type firefly luciferase, in which an amino acid identified based on homology analysis as corresponding with the 421st amino acid (glycine) of the amino acid sequence of the wild-type North American firefly (Photinus pyralis) luciferase has been substituted with a polar amino acid.

Abstract: The present invention relates to luciferase having resistance to a surfactant and a method for measuring intracellular ATP which is characterized in that the luciferase having resistance to a surfactant is used in this method comprising the steps of: a first step wherein ATP is extracted from cells in a sample; a second step wherein light emission is produced by adding a luminescence reagent containing luciferase to the extracted ATP solution; and a third step wherein the light emission is measured.

Abstract: The present invention provides a mutant firefly luciferase consisting of a mutant amino acid sequence derived from the amino acid sequence of a wild-type firefly luciferase by at least substitution (a), (b), or (c) below and having luminescence intensity higher than that of the wild-type firefly luciferase.

Abstract: Provided are methods and compositions useful in detecting protease activity in a sample, as well as methods of identifying agents that modulate protease activity. The methods and compositions provide a modified luciferase polynucleotide sequence and a luciferase polypeptide containing protease recognition sequences, wherein cleavage of the recognition sequence by a protease inhibits luciferase activity. Further provided are methods and compositions for detecting and modulating caspase activity and apoptosis.

Abstract: The present invention provides a polynucleotide or polynucleotides encoding Oplophorus luciferase which is composed of 19 kDa and 35 kDa proteins, or the 19 kDa photoprotein, the recombinant secretional Oplophorus luciferase or the 19 kDa photoprotein encoded by the polynucleotide(s), an expression vector containing the polynucleotide(s) and a host transformed with the vector.

Abstract: The present invention provides a mutant firefly luciferase consisting of a mutant amino acid sequence derived from the amino acid sequence of a wild-type firefly luciferase by at least substitution (a), (b), or (c) below and having luminescence intensity higher than that of the wild-type firefly luciferase.

Abstract: The present invention relates to luciferase having resistance to a surfactant and a method for measuring intracellular ATP which is characterized in that the luciferase having resistance to a surfactant is used in this method comprising the steps of: a first step wherein ATP is extracted from cells in a sample; a second step wherein light emission is produced by adding a luminescence reagent containing luciferase to the extracted ATP solution; and a third step wherein the light emission is measured.

Abstract: The present invention provides a gene construct encoding pH insensitive luciferase for visualizing intracellular information, wherein an intracellular expression activity is higher compared with a gene construct of luciferase derived from a firefly.

Abstract: The present invention provides a polynucleotide or polynucleotides encoding Oplophorus luciferase which is composed of 19 kDa and 35 kDa proteins, or the 19 kDa photoprotein, the recombinant secretional Oplophorus luciferase or the 19 kDa photoprotein encoded by the polynucleotide(s), an expression vector containing the polynucleotide(s) and a host transformed with the vector.

Abstract: A polynucleotide encoding a secreted form of wild type Renilla luciferase. Also provided is a polynucleotide encoding a secreted modified form of wild type Renilla luciferase. Additionally, the polypeptides encoded by the polynucleotides of the present invention and uses of the polynucleotides and polypeptides of the present invention in biological assays. Also, a stable mammalian packaging cell line which produces retroviruses carrying a polynucleotide encoding a secreted Renilla luciferase.

Abstract: Nucleic acid compositions and polypeptides encoding a red-shifted form of firefly luciferase are provided. These red-shifted luciferases are characterized by spectrum of light emission having detectable emissions at 610 nm (luc610), preferably a primary peak at 610 nm. The nucleic acid compositions find use in various systems as a reporter gene, and are of particular interest for use as a reporter with in vivo systems, because of the efficient transfer of red light through tissues. The red-shifted luciferase may be combined in such assays with luciferases emitting at other spectra, in order to monitor multiple processes simultaneously.

Abstract: Described herein is a variant of wild type Gaussia luciferase that catalyzes glow-type emission kinetics suited for high-throughput functional screening applications. Polypeptides, functional fragments, variants, and nucleic acids that encode the enhanced luciferase are further described. One such polypeptide corresponds to wild type Gaussia luciferase with a substitution mutation of I for M at position 43 of the mature peptide. Methods of use, assay systems and kits that contain the polypeptides and/or nucleic acids are further described.

Abstract: The present invention provides a polynucleotide or polynucleotides encoding Oplophorus luciferase which is composed of 19 kDa and 35 kDa proteins, or the 19 kDa photoprotein, the recombinant secretional Oplophorus luciferase or the 19 kDa photoprotein encoded by the polynucleotide(s), an expression vector containing the polynucleotide(s) and a host transformed with the vector. Further, the invention provides a method for producing the recombinant Oplophorus luciferase or the photoprotein. These proteins could be recombinantly produced by culturing the host cell or by in vitro translation system using the recombinant expression vector.

Abstract: Isolated nucleic acid molecules which code for luciferases able to produce the green bioluminescence of Phrixotrhix vivianii and red bioluminescence of Phrixothrix hirtus are described. The nucleic acid molecules and the luciferases encoded thereby can be used in applications such as diagnostic methods and molecular biology tools.

Abstract: The present invention provides a gene construct encoding pH insensitive luciferase for visualizing intracellular information, wherein an intracellular expression activity is higher compared with a gene construct of luciferase derived from a firefly.

Abstract: A modified luciferase protein which is a sensor for molecules including cAMP is provided. The modified luciferase protein includes one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with cAMP.

Abstract: The present invention relates to luciferase having resistance to a surfactant and a method for measuring intracellular ATP which is characterized in that the luciferase having resistance to a surfactant is used in this method comprising the steps of: a first step wherein ATP is extracted from cells in a sample; a second step wherein light emission is produced by adding a luminescence reagent containing luciferase to the extracted ATP solution; and a third step wherein the light emission is measured.

Abstract: Proteins are provided having luciferase activity with lower Km than wild-type luciferases by altering the amino acid residue at position 270 of the wild-type to an amino acid other than glutamate. Greater heat stability than wild-type luciferases while retaining the lower Km is provided by also replacing the glutamate equivalent to that at position 354 of Photinus pyralis luciferase or 356 of Luciola luciferases with an alternative amino acid, particularly lysine and/or the amino acid residue at 215 of Photinus pyralis and 217 of the Luciola species with a hydrophobic amino acid. DNA, vectors and cells that encode for and express the proteins are also provided as are test kits and reagents for carrying out luminescence assays using the proteins of the invention.

Abstract: Described herein is a variant of wild type Gaussia luciferase that catalyzes glow-type emission kinetics suited for high-throughput functional screening applications. Polypeptides, functional fragments, variants, and nucleic acids that encode the enhanced luciferase are further described. One such polypeptide corresponds to wild type Gaussia luciferase with a substitution mutation of I for M at position 43 of the mature peptide. Methods of use, assay systems and kits that contain the polypeptides and/or nucleic acids are further described.