62 results about "Luciferin" patented technology

A method to detect the presence or amount of at least one molecule in a sample which employs a derivative of luciferin or a derivative of a fluorophore is provided.

A method for evaluating sterilization effect of disinfectant by means of ATP bioluminescence includes preparing standard bacterial suspension, processing it with cell ATP release agent Ec and adding luciferase - luciferin reagent to detect out luminous value or filtering it with microhole film in combination with ATP scavenger and detecting out luminous value of filtering film, then carrying out equation calculation and utilizing conversion coefficient K to derive out live bacteria number in suspension for evaluation.

A compound represented by the following general formula (I) or a salt thereof:

[wherein R¹ and R² represent hydrogen atom, a C_1-6 alkyl group, or a group represented by the following formula (A):

[wherein X¹ and X² represent hydrogen atom, or a group represented as —N(R³)(R⁴) (R³ and R⁴ represent hydrogen atom, a C_1-6 alkyl group, a C_1-6 alkylcarbonyl group, or a C_1-6 alkyloxycarbonyl group); and n represents an integer of 1 to 6], provided that R¹ and R² do not simultaneously represent hydrogen atom), which is a novel luciferin derivative that serves as a luciferase substrate.

Methods, devices and systems are provided for detection of adenosine triphosphate (ATP) in samples using the luciferin-luciferase reaction. An aspect of the invention includes a low molarity and low pH composition for use in detecting the presence of ATP. The low molarity and low pH composition can be used in combination with methods for reading, calculating and interpreting luminescence generated by the ATP-luciferin-luciferase reaction. Both the low molarity, low pH composition and the methods for reading, calculating and interpreting luminescence can be used with a single service hygiene monitoring format.

A reagent system comprises a first reagent which includes a high pH phosphate buffer, and a second reagent which includes luciferase, luciferin, a magnesium salt and an enzyme stabilizer. The second reagent has a low pH and a buffer with a pK which is near the optimum pH for activity of luciferase. The reagent system may be used in a process for measuring total adenosine triphosphate (ATP) and/or dissolved extracellular ATP, in a fluid containing microorganisms. The reagent system may also be used in a microbiological remediation or production process.

The present invention relates to:

an isolated or synthesized gene, which encodes a protein comprising the amino acid sequence represented by SEQ ID NO: 2,

an isolated or synthesized gene, which encodes a protein comprising an amino acid sequence comprising a deletion, substitution or addition of one or more amino acids with respect to the amino acid sequence represented by SEQ ID NO: 2 and being capable of regenerating luciferin,

an isolated or synthesized gene, which hybridizes with the complementary strand sequence of a DNA comprising the nucleotide sequence represented by SEQ ID NO: 1 under stringent conditions and encodes a protein capable of regenerating luciferin,

a recombinant DNA, which is characterized in that the above-described isolated or synthesized gene is inserted into a vector DNA,

a transformant or transductant comprising the above-described recombinant DNA, and

a process for producing a protein capable of regenerating luciferin, which is characterized in that the method comprises culturing the above-described transformant or transductant in a medium and collecting therefrom the protein capable of regenerating luciferin.

The invention discloses a DNA fluorescent detection process and its agent box. In the method, target direction DNA is captured through cross-fertilizing between DNA after the capturing detecting probe is coupled to the magnetic particle surface, another nucleotide sequence of target direction DNA is cross-fertilized with semaphore detecting probe modified by luciferin to form sandwich filler structure of magnetic particle-target direction DNA-fluorescence labeling semaphore detecting probe marked, meanwhile competitiveness detecting probe of above capture detecting probe or semaphore detecting probe is added to combine with target direction DNA, finally, water-solubility conjugate high molecule with semaphore multiply function is added to amplify and detect the signal in order to detect the target direction DNA. The invention can improve greatly the selectivity and sensitivity of gene and gene mutation detection without amplifying and increasing nucleic acid and with quick and easy operation, has important meaning in biological detection, morbid early diagnosis and treatment.

The present invention concerns methods of quantifying nucleic acids using a bioluminescence regenerative cycle (BRC). In BRC, steady state levels of bioluminescence result from processes that produce pyrophosphate. Pyrophosphate reacts with APS in the presence of ATP sulfurylase to produce ATP. The ATP reacts with luciferin in a luciferase-catalyzed reaction, producing light and regenerating pyrophosphate. The pyrophosphate is recycled to produce ATP and the regenerative cycle continues. Because the kinetic properties of ATP sulfurylase are much faster than luciferase, a steady state results wherein concentrations of ATP and pyrophosphate and the rate of light production remain relatively constant. Photons are counted over a time interval to determine the number of target molecules present in the initial sample. The BRC process has a controllable dynamic range up to seven orders of magnitude and is sensitive enough to detect a few thousand molecules of target nucleic acid.

A fast detection method of microbe anti- interference includes using large sample method or MPN method to enrich bacteria for microbe culture media sample, processing prepared sample with cell ATP release agent Ec and adding luciferase - luciferin reagent and deinhibitor for ATP luminous detection, confirming that sample is bacteria carrier positive if sample luminous pulse counts CPMs is greater than control sample solution CPMck, checking MPN table to obtain bacteria content in sample. The method can raise detection sensitivity by 1000 times comparing to normal method.

Bioluminescent endoscopy methods and compounds, wherein an anatomical object is examined by means of filling, perfusing, intubating, injecting, or otherwise administering a solution containing a bioluminescent substance or a mixture of luciferin and luciferase which produces bioluminescence, wherein a color or monochrome image of the object is constituted using the images and information based on bioluminescent emitted by the bioluminescent substance. Procedures are demonstrated which allow bioluminescent solutions to be perfused into certain tissue regions, such as but not limited to the common biliary duct, genitourinary tract, gastro-intestinal tract, cardiovascular system and lymphatic system wherein said structures may be conveniently visualized during surgery to avoid damage to these structures. Such images may also be combined with visual light images. Methods of detection of cancer cells using bioluminescence are provided. Preferred embodiments disclosed include membrane permeant coelenterazine analogs.

The present invention provides methods and systems that uses transgenic zebrafish with an easily assessable reporter gene under the control of pollutant-inducible DNA response elements. Transgenic zebrafish, carrying pollution-inducible response elements, are placed in the water to be tested, and the contaminants become bioconcentrated (generally 1,000- to 40,000-fold, relative to the water) in the tissues of the fish thereby activating specific response elements, which up-regulate the LUC reporter gene. Fish are then removed from the test water and placed immediately in a luminometer cuvette and incubated with luciferin. Luciferin is rapidly taken up into the tissues of the fish, oxidized by luciferase, and light is produced. The luminescence is proportional to the environmental concentration of the pollutant (to which the fish had been exposed), which drives the expression of the LUC gene by means of the various DNA motifs. The luminescence is quantitated in the luminometer. In each response element-containing construct, a specific class of polluting chemicals, allowing for differential identification of pollutants in a complex mixture activates the expression of the LUC gene. This assay does not require killing the fish and allows for repeated analysis of the same site with the same fish. The sensitivity of the system can be manipulated by varying the sequence of the response element.

The invention concerns the use of bioluminescence dependent on the reaction (1): luciferin+ATP+O₂+Mg²⁺+luciferase→oxyluciferin+photons for detecting and counting living cells of a given species potentially present in a liquid sample, said use being characterized in that it consists in measuring the total free intracellular adenyl nucleotides (AN) content, expressed in ATP form, of living cells of a given non-viral species, taking into account the fact that the sum of free intracellular ATP, ADP and AMP of said family is constant according to the relationship (2): [AN]=[ATP]+[ADP]+[AMP]=Cte after transforming the free intracellular ATP, ADP and AMP by using myokinase and pyruvate kinase, said measurement being performed (i) without adding ATP and (ii) after adding a known amount of ATP. The invention also concerns a method for detecting and counting cells by ATP-metry.

Provided herein are imaging methods for detecting, diagnosing and imaging pathogenic bacteria or a pathophysiological condition associated therewith using fluorescent, luminescent or colorimetric detection agents, e.g., fluorogenic substrates for bacterial enzymes, caged luciferins and fluorescent proteins, luciferases and enzymes expressed by recombinant bacteria. Signals emitted by the fluorescent, luminescent or colorimetric detection agents in the presence of the bacteria are compared to controls to detect and locate the pathogenic bacteria. Also provided is a method for screening therapeutic agents to treat the pathophysiological conditions by measuring fluorescence or luminescence emitted from the detection agents in the presence and absence of the potential therapeutic agent. In addition, a method for detecting a pathogenic bacteria via PET or SPECT imaging using a positron-emitting or gamma-emitting substrate for a beta-lactamase or other enzyme or protein of the pathogenic bacteria. Further provided are the fluorogenic substrates CNIR-7 or CNIR7-TAT or the radiolabeled substrates.

Transgenic plants are created having incorporated into them a luciferase enzyme gene and a corresponding luciferin substrate gene. These genes are regulated such that for a certain amount of time after dark, these genes are expressed resulting in bioluminescence. Different luciferin/luciferase combinations may be utilized for these transgenic plants, depending on the desired wavelength and the plant species transfected.

Micro-organisms which over-produce ATPase, especially mycoplasmas in a sample of a cell culture, are detected by making use of the ATPase activity to convert cellular or externally added ATP to ADP. The decrease in ATP externally added or the increase or absolute level of ADP produced from cellular ATP can then be detected or measured. For example, if a reagent such as phosphenol pyruvate/pyruvate kinase is added to convert that ADP back to ATP, the resulting high level of ATP can be detected bioluminescently using the reaction: Luciferin+luciferase enzyme+ATP→hv+products.

The invention relates to methods and compositions which utilize the emission of light to monitor changes in microenvironments involving cells. The invention is especially useful for monitoring exocytotic activity such as detecting quantal release of synaptic vesicles. Fusion proteins of Cypridina luciferase and synaptotagmin-I or VAMP/synaptobrevin-2 were targeted to synaptic vesicles and, upon exocytosis, formed light-emitting complexes with luciferin present in the extracellular medium. Photon emissions in the presence of a depolarizing stimulus can be observed with these systems. pH-sensitive mutants of green fluorescent protein are also provided, which are useful for visualizing exocytosis and for imaging and measuring the pH of intracellular compartments.

Provided herein are isolated polynucleotide encoding modified click beetle luciferase polypeptides that have enhanced luminescence and longer wavelength near-infrared signals. The disclosure also relates to near-infrared bioluminescence systems that include said modified click beetle luciferase polypeptides and novel luciferin derivatives, as well as methods of using said modified click beetle luciferase polypeptides and bioluminescence systems.

The invention relates to non-human transgenic animals expressing bioluminescent markers

The invention provides a transgenic mouse whose genome comprises a nucleotide sequence encoding lucif erase, operably linked to murine control sequences whereby the lucif erase is expressed in all cells. Administration of a luciferin substrate to the mouse results in bioluminescence. The control sequence is the Rosa 26 promoter. This mouse is useful to monitor in situ cell growth, differentiation or proliferation