60 results about "Photoprotein" patented technology

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 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 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.

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.

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.

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.

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.

The invention discloses a class of triplet-state intersystem crossing cyanine dyes, a preparation method and application thereof, belongs to the technical field of high-efficiency triplet photosensitizers, and discloses a class of pentamethylcyanine dyes which do not depend on heavy atoms and have high-efficiency intersystem crossing, wherein the dyes have a good application prospect in the fields of photocatalysis, photodynamic therapy and phosphorescence imaging. According to the triplet dye, active groups with rich compatible types and groups with large steric hindrance are constructed on a pentamethylcyanine dye matrix, so that non-radiative transition is limited, and the singlet-triplet energy level difference is smaller. The molecule can stably exist in most of organic solvents, solid states and aqueous solutions, and has excellent active oxygen generation capacity in salt ions, visible light, protein and cells. The molecules can absorb near-infrared light to reach a triplet state, so that electron energy, phosphorescence signals and active oxygen are generated by utilizing the triplet state.