61 results about "Photoprotein" patented technology

A Bioluminescent Paintball 10 includes a shell 12 defining an interior cavity 14, a liquefied substance 16 disposed within the interior cavity 14, a phosphorescent material 18 disbursed throughout the shell 12 for providing a visible “tracing” effect when the bioluminescent paintball 10 is ejected from a paintball discharge device, a neutralizing agent 20 disbursed throughout the liquefied substance 16 for neutralizing calcium disbursed throughout the liquefied substance 16 thereby preventing light emission before the paintball 10 impacts a target, and a photoprotein 22 disbursed throughout the liquefied substance 16 for reacting with calcium disposed upon a target after the bioluminescent paintball 10 impacts the target, thereby rupturing the shell 12 and allowing the liquefied substance 16 to engage the calcium to produce visible light.A paintball 100 includes a shell 102 defining an interior cavity 104, an insoluble coating 106 disposed upon an inner surface 108 of the shell 102, and an aqueous material 110 disposed within the cavity 104 such that the aqueous material 110 engages the insoluble coating 106, thereby preventing the aqueous material 110 from dissolving the shell 102, and promoting the marking of a target via the aqueous material 110 when the paintball 100 forcibly engages the target and ruptures the shell 102. A paintball 200 includes first and second half shell portions 202 and 204 with recesses 206 that receive respective first and second liquids 208 and 210 containing dyes or other marking pigments. The second liquid 210 becomes relatively viscous after being disposed in the second shell portion 204, thereby allowing the second shell portion 204 to be inverted with the second liquid 210 maintaining a constant position in the “up-side down” second shell portion 204 to promote the integral joining of the first and second half shell portions 202 and 204 to form a paintball 200.

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.

The invention provides a dual-color single-photon transverse super-resolution imaging method, and a device thereof. The method has the most important characteristic that fluorescence excitation can be achieved only by adopting a overlapping part of a sensitization beam and an excitation beam to simultaneously sensitize and excite samples labeled with reversible photosensitive fluorescent protein molecules. The method comprises the following steps: fluorescence signals are collected by an objective, pass a sensitization dichroic mirror, an excitation dichroic mirror, a notch light filter and a long-channel light filter, and are focused by a focusing lens to pass a pinhole aperture; fluorescence intensity is measured by an avalanche diode; fluorescence intensity values are recorded by a computer; the computer closes a shutter so as to restore reversible photosensitive fluorescent protein to an excitable state under the illumination of sensitization light; and the computer drives a 3-D translation stage to move the samples labeled with the reversible photosensitive fluorescent protein, so as to achieve 3-D scanning imaging. The method can improve the transverse resolution of super-resolution fluorescence imaging by 1.55 to 2.81 times.

A Bioluminescent Paintball 10 includes a shell 12 defining an interior cavity 14, a liquefied substance 16 disposed within the interior cavity 14, a phosphorescent material 18 disbursed throughout the shell 12 for providing a visible “tracing” effect when the bioluminescent paintball 10 is ejected from a paintball discharge device, a neutralizing agent 20 disbursed throughout the liquefied substance 16 for neutralizing calcium disbursed throughout the liquefied substance 16 thereby preventing light emission before the paintball 10 impacts a target, and a photoprotein 22 disbursed throughout the liquefied substance 16 for reacting with calcium disposed upon a target after the bioluminescent paintball 10 impacts the target, thereby rupturing the shell 12 and allowing the liquefied substance 16 to engage the calcium to produce visible light.

Methods for extending light-emitting time of a solution of a calcium-binding photoprotein that instantaneously emits light by binding to calcium ions are provided. In the light-emitting reaction system of a solution of a calcium-binding photoprotein, a light-emitting reaction is performed in the presence of an anion capable of binding to the calcium ion or the cation that can be substituted for the calcium ion and / or a cation that can bind to the calcium-binding site of the calcium-binding photoprotein with a lower affinity than the calcium ion or the cation that can be substituted for the calcium ion without activating the calcium-binding photoprotein.

The invention provides large stokes displacement fluorescent protein CyOFP and application of the fluorescent protein to microimaging and highly-sensitive living creature light-emitting imaging. The fluorescent protein is obtained through mNeptune site directed mutagenesis, the 1-230 positions of the amino acid sequence of the fluorescent protein sequentially correspond to 4-234 positions shown by the SEQ ID No: 2, or the fluorescent protein can be further obtained by conducting gene synthesizing on the fluorescent protein DNA segment and expression. The invention further provides a novel BRET system and fusion protein Antares obtained through system optimization. The CyOFP can be stimulated by blue light, and a quite high quantum yield is achived; the CyOFP and the enhanced green fluorescent protein (EGFP) are jointly stimulated by monochromatic light at the same time in a two-photon mode, and the resolution ratio of two-photon imaging is raised; a novel BRET system formed together with fluorescent protease NanoLuc greatly improves the depth and sensitivity of living creature imaging.

The invention relates to multiple improved orange / red fluorescence protein. By modification of amino acid sequence of red fluorescence protein DsRed from Discosoma sp., the obtained novel fluorescence protein has different characteristics from a wild type. A brighter monomer structure orange fluorescence protein similar to mOrange2 is obtained, and the fluorescent brightness is 2.0 times higher than fluorescent brightness of the mOrange2. According to the invention, a novel purple red fluorescence protein is also obtained, and the fluorescence protein has specific maximum excitation wavelength and maximum emission wavelength. The invention also relates to research of the improved fluorescence protein applied in multi-fields. Positioning, tracing, functional display and the like of target proteins in cells, subcells and in-vivo tissues can be analyzed.

The invention belongs to the technical field of nano materials and particularly relates to a method for preparing fluorescent magnetic nanoparticles with streptavidin combination function. The method comprises the following specific steps of: respectively connecting a Strep II label gene to N ends of alpha and beta sub-gene apoprotein genes to obtain Strep II-apcA and Strep II-apcB gene segments; embedding the Strep II-apcA gene segment to the downstream of a 6*His gene, and cloning to one expression vector together with a phycobilin biosynthetic enzyme gene; embedding the Strep II-apcB gene segment to the downstream of the 6*his gene, cloning to another vector together with a chromphore lyase gene, simultaneously converting the two expression vectors into escherichia coli, screening engineering bacteria, separating and purifying the engineering bacteria by protein, and oscillating and mixing the purified double-label recombinant APC fluorescent protein and zinc ion modified superparamagnetism silicon shell nanoparticles to obtain the fluorescent magnetic nanoparticles with the streptavidin combination function. The obtained double-label recombinant protein can biologically combine streptavidin without being modified chemically.

The object of the present invention is to provide a method for screening a substance involved in a metabolic shift of skeletal muscle, and a kit for screening a substance involved in a metabolic shift of skeletal muscle.The object can be solved by a muscle stem cell or myoblast comprising at least one myosin-heavy chain fusion gene selected from the group consisting of a myosin-heavy chain I fusion gene wherein a myosin-heavy chain I gene and a fluorescent protein or photoprotein gene are fused, a myosin-heavy chain IIa fusion gene wherein a myosin-heavy chain IIa gene and a fluorescent protein or photoprotein gene are fused, a myosin-heavy chain IId / x fusion gene wherein a myosin-heavy chain IId / x gene and a fluorescent protein or photoprotein gene are fused, and a myosin-heavy chain IIb fusion gene wherein a myosin-heavy chain IIb gene and a fluorescent protein or photoprotein gene are fused.

This invention is to provide a photoprotein which binds with a ligand specific for a substance to be detected at a binding ratio of 1:1 such that the luminescence activity is not reduced by binding with the ligand, a conjugate comprising the luminescent photoprotein and ligand, and a substance detection method which employs the conjugate as a marker. A calcium-binding photoprotein is produced having cysteine residue introduced within the 4th amino acid residue from the amino-terminus. A conjugate is formed by binding a ligand specific for a substance to be detected to the calcium-binding photoprotein, in a binding ratio of 1:1, via the introduced cysteine residue. The conjugate may be utilized as a marker for a substance to be detected.

The present invention relates to a method to produce activated split-polypeptide fragments that on reconstitution immediately forms an active protein. The method relate to real-time protein complementation. Also encompassed in the invention is a method to split and produce split-fluorescent proteins in an active state which produce a fluorescent signal immediately on reconstitution. The present application also provides methods to detect nucleic acids; non-nucleic acid analytes and nucleic acid hybridization in real-time using the novel activated split-polypeptide fragments of the invention.

The present invention relates to photoproteins with enhanced bioluminescence obtained by mutagenesis of clytin, to their use as intracellular calcium indicators and in cell-based assays.

There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analogue showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analogue suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.

The invention discloses a far infrared fluorescent protein which comprises an amino acid sequence of a BDFP near infrared fluorescent protein and comprises mutants at amino acids of the 30th, 101st, 107th, 109th, 113th, 130th, 143rd, 151st and 163rd loci. The far infrared fluorescent protein does not comprise mutants at amino acids of the 35th, 120th and 122nd loci. The amino acid sequence of theBDFP near infrared fluorescent protein is as shown in SEQ ID NO: 2. The invention also discloses a fused fluorescent protein of the far infrared fluorescent protein. The invention further discloses anucleic acid which encodes the far infrared fluorescent protein or the fused fluorescent protein thereof and a carrier including the nucleic acid.

The invention relates to methods for the quantitative optical analysis of the intracellular concentration of the cyclic nucleotides cGMP and cAMP, said methods using cell lines which express a combination of certain CNG channels, a calcium-sensitive photoprotein, and different target proteins for which modulators are to be found, in a recombinant manner. The cell lines modified in this way are suitable for high-throughput screening (HTS and uHTS) and can be used to identify medicaments which influence the activity of receptors or enzymes participating in the composition or decomposition of the cyclic nucleotides cGMP and cAMP.

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.

The invention discloses a plasmid for detecting and screening circulating tumor cells and a detecting and screening method using the same, and relates to the technical field of detecting and screening of cells. By the detecting and screening method, the plasmid for detecting and screening the circulating tumor cells can screen surface antigens of the circulating tumor cells in a targeted manner, can screen nickel magnetic beads in combination with a his tag, and is good in specificity and high in sensitivity. By the circulating tumor cell detecting and screening method, a single-chain variable fragment scFvHer2 of an antibody is connected with a biological light-emitting protein sequence Rluc extracted from sea pansy, the plasmid with the his tag transfects 293ft cells to obtain fusion protein, and the fusion protein can target the surface antigens of the circulating tumor cells and is transformed into an amplified signal for detecting and screening the circulating tumor cells; the detecting and screening method is safe in operation, low in cost, high in specificity and high in sensitivity; the detecting and screening method can be applied to detection and screening of the CTCs (circulating tumor cells) in peripheral blood of a patient suffering from breast cancer, and the screened CTCs are applied to primary culture and gene or protein detection.

A modified bioluminescent system comprising a fluorescent molecule covalently linked with a photoprotein, wherein said link between the two proteins has the function to stabilize the modified bioluminescent system and allowing the transfer of the energy by Chemiluminescence Resonance Energy Transfer (CRET).

The present invention provides a fluorescent protein, and a preparation method and an application thereof, and relates to the technical field of biology. The novel red large Stokes shift fluorescent protein is screened out, can be combined with a green fluorescent protein in a single photon and multi-photon microscopy imaging together to be effectively stimulated by monochromatic light for multi-color imaging, has good monomerity, and also has greatly improved light stability. At the same time, the fluorescent protein can be used as a donor to be combined with an acceptor luciferase to form anew highly sensitive bioluminescence resonance energy transfer (BRET) system in organisms. When detecting intramolecular interactions in cells, a dynamic range of the bioluminescence resonance energytransfer system is 2.27 times higher than that of a traditional bioluminescence resonance energy transfer system. The application of the provided protein improves efficiency of the BRET system and improves the dynamic range of the bioluminescence resonance energy transfer system for detecting the intramolecular interactions.

There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analogue showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analogue suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.

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.

The present invention relates to photoproteins with enhanced bioluminescence obtained by mutagenesis of clytin, to their use as intracellular calcium indicators and in cell-based assays.

The invention relates to an application of a fluorescent dye with an intramolecular switch in super-resolution imaging. The dye is pentamethine cyanine with the intramolecular switch. The switch is in a closed-loop state in most organic solvents, in a solid state and in a wide pH range from acidity to alkalinity of an aqueous solution, and molecules are protected from photobleaching. Under an imaging condition, a small amount of molecules can be subjected to ring opening under the action of protons, salt ions, laser, proteins and other substances in cells, and closed-loop molecules still account for most of the molecules. The molecules are low in ring-opening proportion, high in ring-opening molecule brightness, high in light stability and suitable for super-resolution imaging.

A photoresist stripping composition comprising an organic amine and a method is provided. The photoresist stripping composition comprising an organic amine having the following formula (1).

The invention discloses a molecular probe for tracking oxytocin. A fusion protein comprises oxytocin, a connecting ring, fluorescent protein 1, an enzyme cleavage site, neurophysin and fluorescent protein 2. The molecular probe of the invention can track the oxytocin and the neurophysin, and the molecular probe can also explore the molecular structure relationship between the oxytocin and the neurophysin.

A fluorescent protein (bFP) having chemiluminescence activity is a complex composed of the apoprotein of a calcium-binding photoprotein, coelenteramid or an analog thereof, and calcium ions or divalent or trivalent ions that can be substituted for the calcium ions. In the complex, the ratio of the number of molecules of the apoprotein to that of the coelenteramid is 1:1 and the ratio of the number of molecules of the apoprotein to that of the divalent or trivalent ions is 1:1 to 1:4. The fluorescent protein is used as a marker because it catalyzes luminescence of coelenterazine and has fluorescence capability. Removal of calcium ions etc. from this fluorescent protein (bFP) having luminescence activity provides a novel fluorescent protein (gFP). Mixing this gFP with the coelenterazine provides a calcium-binding photoprotein, which emit light instantaneously, enabling use as a marker.