32 results about "Luminescent Proteins" patented technology

A method to detect proteasome activity in permeabilized cells, and optionally in a multiplex assay to detect presence or amount of at least one molecule for a different enzyme-mediated reaction, is provided.

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.

The present invention describes a method for detecting biomolecular interactions said method comprising: (a) selecting an appropriate reporter molecule selected from the group consisting of a protein, a fluorescent protein, a luminescent protein and a phosphorescent protein; (b) effecting fragmentation of said reporter molecule such that said fragmentation results in reversible loss of reporter function; (c) fusing or attaching fragments of said reporter molecule separately to other molecules; followed by (d) reassociation of said reporter fragments through interactions of the molecules that are fused to said fragments; and (e) detecting said biomolecular interactions by reconstitution of activity of the reporter molecule with the proviso that said protein is not ubiquitin.

The instant invention describes a method for detecting protein-protein interactions in living organisms and / or cells, said method comprising: (a) synthesizing probe protein fragments from a protein which enables fluorescent or luminescent detection by dissecting the gene coding for the fluorescent or luminescent protein into a least two fragments; (2) constructing fusion proteins consisting of the probe protein fragments linked to protein domains that are to be tested for interactions; (3) coexpressing the fusion proteins; and (d) detecting reconstitution of the fluorescence or luminescence signal.

There is provided a novel luminescence measuring apparatus that enables measuring a luminescence amount or a luminescence intensity from plural living biological samples, such as tissues, cells, and biological individuals, into which a gene expressing a luminescent protein has been introduced, individually for the respective biological samples. The inventive luminescence measuring apparatus comprises an image acquisition portion that acquires a luminescence image of biological samples; a measurement region determination portion that determines at least one measurement region in the luminescence image; and a luminescence measurement portion that measures the luminescence amount or luminescence intensity in the determined measurement region; thereby measuring the luminescence amount or luminescence intensity individually by the biological sample(s) included in the measurement region. Irrespective of an efficiency of transfer of a luminescent protein gene into a cell, etc., it becomes possible to execute a luminescence measurement successfully while observing only biological samples having a gene expression.

A method for monitoring the differentiation state of a stem cell, includes a step (A-1) of culturing a stem cell into which a fusion gene of a promoter region of a differentiation state detection marker gene and a luminescent protein-coding gene is introduced, a step (A-2) of culturing the stem cell under a differentiation-inducing condition after the step (A-1), and a step (A-3) of capturing images of luminescence emitted by expression of the luminescent protein-coding gene in the stem cell, over at least a given period of the steps (A-1) to (A-2).

The present invention relates to fluorescent or luminescent probe reagents for measuring oxidative stress in a cell or an organism. Examples of the probe reagents include: a fluorescent or luminescent protein and a marker protein; a fluorescent or luminescent protein, a marker protein, and a regulatory factor; or a fluorescent or luminescent protein, a marker protein, a cleavage sequence, and a regulatory factor. In the probe reagents, the marker protein makes it possible to detect the oxidative stress caused by reactive oxygen species and comprises a regulatory factor-binding site and a ubiquitin-binding site; and the regulatory factor is a protein making it possible to regulate degradation of the marker protein in response to the reactive oxygen species. The present invention also relates to a method of measuring oxidative stress in a cell or an organism, or a method of screening a substance which suppresses or promotes the oxidative stress in a cell or an organism by using the probe reagent.

The instant invention describes a method for detecting protein-protein interactions in living organisms and / or cells, said method comprising: (a) synthesizing probe protein fragments from a protein which enables fluorescent or luminescent detection by dissecting the gene coding for the fluorescent or luminescent protein into a least two fragments; (2) constructing fusion proteins consisting of the probe protein fragments linked to protein domains that are to be tested for interactions; (3) coexpressing the fusion proteins; and (d) detecting reconstitution of the fluorescence or luminescence signal.

The invention relates to a chemical luminescent protein chip, a kit and a detection method for detecting the fucose index of seroglycoid, and belongs to a protein detection technology. The chemical luminescent protein chip is characterized in that a substrate slide glass of the protein chip at least comprises a detection subregion, wherein one detection subregion is used for detecting one serum sample; two detection spot regions and one line of contrast spot regions are arranged in the detection subregion; a detection spot formed by a specific antibody of fixed alpha fetoprotein is arranged in one detection spot region, and a detection spot formed by fixed lens culinaris agglutinin is arranged in the other detection spot region; a contrast spot formed by fixed bovine serum albumin is arranged in each contrast spot region; and the concentrations of substances on all the detection spots in the same detection spot region are equal. The protein chip, the kit and the method provided by the invention can be used for accurately detecting the fucose index of the seroglycoid with high throughput, and have the advantages of high sensitivity, time saving, convenience, economy and the like in clinical application.

A method for photodynamic therapy treatment of cancerous cells and tissue is provided. The method comprises administering tumor-trophic cells expressing a luminescent protein to a subject. A photosensitizing agent is then separately administered to the subject, followed by an optional iron chelator. On the day of treatment, a luminogenic substrate corresponding to the luminescent protein is administered to the subject. The substrate reacts with the luminescent protein in the vicinity of the cancerous tissue to produce light which activates the photosensitizing agent resulting in the selective destruction of the cancerous tissue.

The invention provides a method for preparing a pre-staining luminescent protein marker. The method comprises the following steps: acquiring a protein A gene, a protein G gene, an MBP (Myelin Basic Protein) gene and a heavy chain constant region gene (CH gene) of a mouse and rabbit derived antibody, and cutting and combining the genes according to sizes of different proteins in a pre-staining luminescent protein marker; expressing the proteins, purifying, and mixing proteins of different molecule weights according to a certain ratio so as to obtain a luminescent protein marker; mixing the prepared luminescent protein marker with the pre-staining luminescent protein marker according to different ratios. The pre-staining luminescent protein marker prepared by using the method is capable of recognizing primary antibodies or secondary antibodies of different sources, binding sites of proteins are increased, light signals are intensified, not only are general positions of proteins indicated in the electrophoresis process and after membrane transfer, but also precise positions of the proteins can be indicated on X-ray films, no extra pre-staining marker channels are needed, no extra antibodies are needed, and the luminescent proteins can be combined with IgG or anti-rabbit and anti-mouse secondary antibodies derived from species such as human beings, rats, mice and rabbits.

A method for monitoring differentiation into cardiac muscle cells includes keeping, in an alive state, cells into which a reporter gene of luminescent protein configured to vary in luminescence intensity according to an expression of myocardial differentiation marker gene is introduced. The method includes acquiring a luminescence image as a still image by imaging light emitted from the cells in a light shielding state. The method includes acquiring sequential images with illuminating the cells. The method includes associating biological information obtained from the sequential images with biological information obtained from the still image.

This invention provides a novel ratiometric bioluminescent sensor and methods of use thereof. The bioluminescent sensor comprises two luminescent proteins that exhibit different characteristics associated a biological condition, and thereby illuminates differently in response to the biological condition. A ratio between the luminescence of the two luminescent proteins indicates a change in the biological condition. The bioluminescent sensor of the invention can be used to image an oxidative stress or its associated conditions, including a programmed cell death.

The present invention relates to a systems and methods of using expression of one or two luminescent proteins in a plant root cell to visualize plant root structure as well as to determine how stressors affect gene expression in plant roots while maintaining the natural soil habitat

The present invention relates to a process of preparing a rubber-like material containing a protein immobilized therein as well as a corresponding rubber-like material the process comprising the stepsof (a) mixing a protein a branched polymer such as trimethylolpropane ethoxylate and a linear polymer such as poly( ethylene oxide) in an aqueous solution to form a gel and (b) drying the gel to obtain a rubber-like material containing the protein immobilized therein wherein the branched polymer comprises at least three polymeric branches bound to a central branching unit. The rubber-like material allows the immobilization and stabilization of a wide range of different proteins including luminescent proteins as well as enzymes and can particularly advantageously be used as down-converting material for light-emitting diodes (LEDs) for diagnostic applications and in bioreactors.

An excellent protein stabilizer is provided, which has the following effects: (1) low probability with contamination of pathogens, (2) the effect of stabilization on photoproteins, and (3) minimization of loss of activity under lyophilizing conditions. A peptide from fish is used as the active ingredient for the protein stabilizer.

Provided are a procedurally simple and safe method and kit for measuring the concentration in a biological sample of a physiologically active substance which binds to a receptor that causes a change in intracellular cAMP concentration. By providing a composition including genetically-modified cells which show co-expression of a receptor that causes a change in intracellular cAMP concentration, a cyclic nucleotide-responsive calcium channel, and aequorin luminescent protein, it is possible to measure how much of a physiologically active substance which binds to said receptor is included in a biological sample.

The present invention relates to fluorescent or luminescent probe reagents for measuring oxidative stress in a cell or an organism. Examples of the probe reagents include: a fluorescent or luminescent protein and a marker protein; a fluorescent or luminescent protein, a marker protein, and a regulatory factor; or a fluorescent or luminescent protein, a marker protein, a cleavage sequence, and a regulatory factor. In the probe reagents, the marker protein makes it possible to detect the oxidative stress caused by reactive oxygen species and comprises a regulatory factor-binding site and a ubiquitin-binding site; and the regulatory factor is a protein making it possible to regulate degradation of the marker protein in response to the reactive oxygen species. The present invention also relates to a method of measuring oxidative stress in a cell or an organism, or a method of screening a substance which suppresses or promotes the oxidative stress in a cell or an organism by using the probe reagent.

Animal food comprises material which improves and / or optimizes the expression of luminescent proteins in animals. The animals may naturally express luminescent proteins or they may be transgenic animals transfected with a non-endogenous luminescent gene. The animal has a predetermined proteome and the luminescent protein has a predetermined composition of amino acids. The animal food has a base food that is fortified with one or more amino acids which are over-represented in the luminescent protein compared to the amino acids present in the animal proteome. The animal food may further comprise caloric material which increases the cellular metabolism of the animal thereby increasing the expression of the luminescent protein. The animal food may also include inducing material which induces expression of an inducible luminescent gene present in the animal. The animal food may comprise amino acids which are present in, or improve production of, at least one animal tissue in which the luminescent protein is present. The animal food may also comprise a carotinoid and / or a luminescent material.

The invention discloses a mechanical hybrid rice seed production method utilizing the transgenic technology of chloroplasts. The method is characterized by transforming the luminescent protein genes of the rice into the chloroplasts of restorer lines of the hybrid rice, wherein the restorer lines have the characteristic of maternal inheritance of fluorescent proteins and the pollens do not carry transgenes, therefore, the hybrids mated by the restorer lines are non-transgenic, proportionally sowing the male and female parents in a mixed manner during hybrid rice seed production, after becoming mature through field regular management, mechanically reaping the seeds and sorting the reaped seeds into non-fluorescing hybrids and fluorescing restorer lines through photoelectric sorting. The mechanical seed production technology not only greatly lightens the labor intensity of the farmers, reduces the operation procedures and lowers the production cost of the seeds but also ensures that the non-transgenic hybrid rice seeds are produced by utilizing the transgenic technology of chloroplasts.

The invention belongs to the technical field of soil detection, and discloses a method for detecting ecotoxicity of heavy metal contaminated soil by using luminous earthworms. The method comprises thefollowing steps: detecting a soil sample by using a soil heavy metal detection device; obtaining the types of the heavy metals contained in the soil of the to-be-detected area; acquiring eisenia fetida from alternanthera philoxeroides, carrying out purification treatment on the eisenia fetida, and removing impurities in the body; measuring an initial fluorescence value of an earthworm body; putting the earthworms into the soil sample for cultivation; collecting the eisenia fetida, secreting mucus, and performing fluorescence value detection to obtain an actual fluorescence value; and calculating to obtain the ecotoxicity of the heavy metal contaminated soil. A principle that the heavy metal inhibits luminescent proteins in the luminous earthworms is utilized, fluorescence values of the mucus generated before and after activity of the luminous earthworms in soil are detected, an average light inhibition rate is calculated, the consumed time is shorter, and the accuracy is higher.

An excellent protein stabilizer is provided, which has the following effects: (1) low probability with contamination of pathogens, (2) the effect of stabilization on photoproteins, and (3) minimization of loss of activity under lyophilizing conditions. A peptide from fish is used as the active ingredient for the protein stabilizer.

Provided are a procedurally simple and safe method and kit for measuring the concentration in a biological sample of a physiologically active substance which binds to a receptor that causes a change in intracellular cAMP concentration. By providing a composition including genetically-modified cells which show co-expression of a receptor that causes a change in intracellular cAMP concentration, a cyclic nucleotide-responsive calcium channel, and aequorin luminescent protein, it is possible to measure how much of a physiologically active substance which binds to said receptor is included in a biological sample.

The present invention provides a method of bioluminescence-driven optogenetic control of excitable cells. The excitable cell expresses a light-gated ion channel, and a luminescent protein can be expressed either in the excitable cell or in another cell proximal to the excitable cell. The methods of the invention can be used to desynchronize local activity of excitable cells in a mammalian tissue. The methods of the invention can be used to treat a disease or condition in a mammal, the disease or condition being related to bursting. The disease or condition can be Parkinson's disease, epilepsy, a sleep disorder, or a sensory-related disease or condition (e.g., attention deficit disorder or pain). The invention also provides a conjugate of containing a voltage-gated ion channel and a luminescent protein.

The invention belongs to the technical field of soil detection, and discloses a method for detecting the ecological toxicity of heavy metal-contaminated soil by using luminescent earthworms. The method for detecting the ecological toxicity of heavy metal-contaminated soil by using luminescent earthworms includes: using a soil heavy metal detection device to detect soil samples , to obtain the types of heavy metals contained in the soil of the area to be detected; obtain Eisenia chinensis from water peanuts, purify Eisenia chinensis to remove impurities in the body; measure the initial fluorescence value of the earthworm body; put the earthworm into the soil sample Cultivate in medium; collect Eisenia chinensis, and detect the fluorescence value after secreting mucus to obtain the actual fluorescence value; calculate the ecological toxicity of heavy metal polluted soil. The present invention utilizes the principle that heavy metals suppress photoproteins in the body of luminescent earthworms, detects the fluorescence value of the mucus produced before and after activities in the soil, and calculates the average light suppression rate, which takes less time and has higher accuracy.

The present invention provides a method of bioluminescence-driven optogenetic control of excitable cells. The excitable cell expresses a light-gated ion channel, and a luminescent protein can be expressed either in the excitable cell or in another cell proximal to the excitable cell. The methods of the invention can be used to desynchronize local activity of excitable cells in a mammalian tissue. The methods of the invention can be used to treat a disease or condition in a mammal, the disease or condition being related to bursting. The disease or condition can be Parkinson's disease, epilepsy, a sleep disorder, or a sensory-related disease or condition (e.g., attention deficit disorder or pain). The invention also provides a conjugate of containing a voltage-gated ion channel and a luminescent protein.

The invention relates to luminescent proteins, nucleic acids encoding same, compositions and combinations comprising the proteins, and methods using the proteins, nucleic acids, compositions and combinations. In particular, a luminescent protein is provided comprising oncomodulin in which a salt bridge has been introduced to provide greater stability. The protein may be used as a luminescent marker in, for example, luminescent items, immunoassays, and fluorescent energy transfer assays.