223 results about "Yellow fluorescent protein" patented technology

The disclosure provides proteins that can be used to determine the redox status of an environment (such as the environment within a cell or subcellular compartment). These proteins are green fluorescent protein (GFP) variants (also referred to as redox sensitive GFP (rosGFP) mutants), which have been engineered to have two cysteine amino acids near the chromophore and within disulfide bonding distance of each other. Also provided are nucleic acid molecules that encode rosGFPs, vectors containing such encoding molecules, and cells transformed therewith. The disclosure further provides methods of using the rosGFPs (and encoding molecules) to analyze the redox status of an environment, such as a cell, or a subcellular compartment within a cell. In certain embodiments, both redox status and pH are analyzed concurrently.

Nucleic acid compositions encoding non-aggregating chromo / fluoroproteins and mutants thereof, as well as the encoded proteins, are provided. The proteins of interest are polypeptides that are non-aggregating colored and / or fluorescent proteins, where the non-aggregating feature arises from the modulation of residues in the N-terminus of the protein and the chromo and / or fluorescent feature arises from the interaction of two or more residues of the protein. Also provided are fragments of the subject nucleic acids and the peptides encoded thereby, as well as antibodies to the subject proteins and transgenic cells and organisms. The subject protein and nucleic acid compositions find use in a variety of different applications. Finally, kits for use in such applications, e.g., that include the subject nucleic acid compositions, are provided.

This invention provides tandem fluorescent protein construct including a donor fluorescent protein moiety, an acceptor fluorescent protein moiety and a linker moiety that couples the donor and acceptor moieties. The donor and acceptor moieties exhibit fluorescence resonance energy transfer which is eliminated upon cleavage. The constructs are useful in enzymatic assays.

A method of screening multiply transformed / transfected cells to identify those cells expressing at least two peptides or proteins of interest. The method comprising: 1. Simultaneously or sequentially transforming a cell with at least two different expression cassettes in which the gene of interest is linked via an IRES to a fluorescent marker gene. Each marker gene is different. 2. Providing conditions in which expression of the genes will occur. 3. Identifying cells expressing proteins by detecting the different fluorescent signals.

Isolated and purified nucleic acids encoding green fluorescent proteins from Renilla reniformis and the green fluorescent protein encoded thereby are also provided. Mutants of the nucleic acid molecules and the modified encoded proteins are also provided. Compositions and combinations comprising the green fluorescent proteins and / or the luciferase are further provided.

The invention provides protein subcellular localization assays using split fluorescent protein systems. The assays are conducted in living cells, do not require fixation and washing steps inherent in existing immunostaining and related techniques, and permit rapid, non-invasive, direct visualization of protein localization in living cells. The split fluorescent protein systems used in the practice of the invention generally comprise two or more self-complementing fragments of a fluorescent protein, such as GFP, wherein one or more of the fragments correspond to one or more beta-strand microdomains and are used to “tag” proteins of interest, and a complementary “assay” fragment of the fluorescent protein. Either or both of the fragments may be functionalized with a subcellular targeting sequence enabling it to be expressed in or directed to a particular subcellular compartment (i.e., the nucleus).

The invention relates to proteins or polypeptides that comprise intramolecular dimers of fluorescent protein monomers. More specifically, the invention relates to recombinant polypeptides comprising a monomer of a fluorescent polypeptide, a linker peptide, and a second monomer of that fluorescent polypeptide, where the monomers form an intramolecular dimer. The invention also relates to nucleic acids encoding Intramolecular Dimer Fluorescent Proteins (IDFPs) and vectors comprising such nucleic acids. The invention further relates to methods of making IDFPs and methods of using them. IDFPs are, useful in any application suited for fluorescent proteins and are particularly useful in applications in which more than one fluorescent protein sharing complementary dimerization interfaces is present in the same mixture or is expressed in the same cell, because IDFPs do not form heterodimers.

The present invention provides nucleic acid molecules encoding a fluorescent and chromo-proteins and mutants, variants and derivatives thereof, as well as proteins and peptides encoded by these nucleic acids. The nucleic acid molecules and proteins of interest are isolated from non-Aequorea Hydrozoa species. The proteins of interest include yellow fluorescent protein, phiYFP, from Phialidium sp., green fluorescent protein hydr1GFP and purple chromoprotein, hm2CP from hydroid medusae of sub-order Anthomedusae. Also of interest are proteins that are substantially similar to, or derivatives, or homologues, or mutants of, the above-referenced specific proteins. Also provided are fragments of the nucleic acids and the peptides encoded thereby, as well as antibodies specific to the proteins and peptides of the invention. In addition, host-cells, stable cell lines and transgenic organisms comprising above-referenced nucleic acid molecules are provided. The subject protein and nucleic acid compositions find use in a variety of different applications and methods, particularly for labeling of biomolecules, cell or cell organelles. Finally, kits for use in such methods and applications are provided.

The disclosure provides proteins that can be used to determine the redox status of an environment (such as the environment within a cell or subcellular compartment). These proteins are green fluorescent protein (GFP) variants (also referred to as redox sensitive GFP (rosGFP) mutants), which have been engineering to have two cysteine amino acids near the chromophore and within disulfide bonding distance of each other. Also provided are nucleic acid molecules that encode rosGFPs, vectors containing such encoding molecules, and cells transformed therewith. The disclosure further provides methods of using the rosGFPs (and encoding molecules) to analyze the redox status of an environment, such as a cell, or a subcellular compartment within a cell. In certain embodiments, both redox status and pH are analyzed concurrently.

A system for optical tomography includes an apparent light source adapted to project excitation light toward a specimen having fluorescent proteins therein, wherein the excitation light enters the specimen becoming intrinsic light within the specimen, wherein the intrinsic light is adapted to excite fluorescent light from the fluorescent proteins, and wherein the intrinsic light and the fluorescent light are diffuse. A method of optical tomography includes generating the excitation light with the apparent light source, wherein the intrinsic light and the fluorescent light are diffuse.

The invention discloses a resin embedding method for biological tissues marked by fluorescent protein. The resin embedding method comprises the following steps: (1), fixing the biological tissues marked by the fluorescent protein in a chemical fixing manner to obtain the fixed biological tissues marked by the fluorescent protein; (2), replacing the fixed biological tissues marked by the fluorescent protein with an organic solvent, so that the biologic tissues are dewatered to obtain dewatered biological tissues marked by the fluorescent protein; (3), performing embedding agent permeation treatment on the dewatered biological tissues marked by the fluorescent protein to obtain embedding-agent monomer-filled biological tissues marked by the fluorescent protein; (4), enabling the embedding agent to generate a polymerization reaction to obtain a resin-embedding sample of the biological tissues marked by the fluorescent protein; (5), soaking the resin-embedding sample in an alkaline solution, and adjusting the pH value to be 8-12. The resin embedding method for the biological tissues marked by the fluorescent protein can be suitable for biologic tissue samples of various sizes, the fluorescence intensity is high, the imaging effect is good, and the samples have excellent cutting performance.

The invention relates to a white light LED (light emitting diode) with high color rendering index, high light efficiency and low color temperature and a preparation method thereof. The white light LED comprises a support, an LED blue wafer and a fluorescent powder layer, wherein the LED blue wafer is fixed on the support; the fluorescent powder layer is coated on the LED blue wafer; and the fluorescent powder is a mixture of yellow fluorescent powder, red fluorescent powder, green fluorescent powder and an adhesive. The white light LED provided by the invention has the advantages of high color rendering index, high light efficiency and low color temperature.

A protein biosensor having increased signal intensity to sugar concentration and the use thereof. A fluorescent indicator protein is described, in which fluorescent proteins of different emission ranges are fused to both ends of a binding protein undergoing a conformational change due to binding of sugars, such that a change in the concentration of various sugars involved in intracellular metabolism, e.g., maltose, can be detected by measuring a change in emission intensities, caused by FRET (fluorescence resonance energy transfer). A method for detecting a change in the concentration of sugars using such fluorescent indicator protein is also described. The fluorescent indicator protein has excellent signal intensity enabling precision measurement of intracellular concentration of various sugars. In one implementation, high signal intensity fluorescent indicator proteins can be prepared from proteins otherwise used as biosensors, to enable them to be more widely useful.

The invention relates to a fluorescent agent color wheel and a light source system using the fluorescent agent color wheel. The fluorescent agent color wheel at least comprises glass, a first fluorescent agent and a first light filtering sheet. The glass is provided with a first segment. The first fluorescent agent coats the first segment. First wave band light is converted into second wave band light in terms of structure. The first light filtering sheet is arranged to be adjacent to the first segment and used for filtering part of first light beams of the first wave band light and the second wave band light so that a second light beam of the second wave band light emits out by penetrating through the first light filtering sheet. The second wave band light is yellow and the first fluorescent agent is a yellow fluorescent agent. The first filtering sheet is a red light filtering sheet. Thus, the manufacturing cost is effectively reduced, the luminance of red light is improved along with boosting of drive currents, therefore, the overall luminance of the light source system is improved and image quality is modified.

The present invention is directed to a rapid, quantitative screening procedure of γ-aminobutyric acid (GABA)-mediated halide transport in cells with the use of a conventional 384-well fluorescence plate reader (FLIPR). The halide sensor is a novel yellow fluorescent protein (YFP) with mutations at positions 46 / 148 / 152 that exhibits bright fluorescence at 37° C.

The invention discloses a method for separating and converting rape protoplast. The method comprises the following steps: 1, selecting rape cotyledons or true leaves, and removing lower epidermis; 2, putting into enzymolysis liquid for carrying out enzymolysis, wherein the enzymolysis liquid is prepared from the following formula components: 1wt% of cellulase, 0.3wt% of macerozyme, 0.5M of mannitol, 20mM of KCl, 20nM of MES with a pH value of 5.7, 10mM of CaCl2 and 0.1wt% of BSA; 3, adding an isovolumetric W5 solution into a mixed solution obtained by means of enzymolysis, carrying out low speed centrifugation for removing supernatant, adding W5 for washing and resuspending, adding resuspended liquid into 20wt% of a sucrose solution for purifying, horizontally centrifuging, and taking supernatant green protoplast liquid; 4, centrifuging for removing the W5 solution, and resuspending protoplast precipitate by using an MMG solution; 5, converting the protoplast. The protoplast separating and converting method is established in rape for the first time; the rape protoplast is used as a receptor for converting, so that yellow fluorescent protein (YFP), luciferase (Luc) and renilla reniformis luciferase (Ren) are successfully expressed.

The invention is an LED device able to regulate color temperature, mainly additionally providing at least a red LED cell and at least a green LED cell outside a white LED device comprising blue LED cell and yellow fluorescent powder coating, to change white light color temperature, where the yellow fluorescent powder coating can only cover the blue LED cell, or additionally cover the red and green LED cells. Additionally, the blue, red and green LED cells can be separately driven to further improve color temperature.

A system for optical tomography includes an apparent light source adapted to project excitation light toward a specimen having fluorescent proteins therein, wherein the excitation light enters the specimen becoming intrinsic light within the specimen, wherein the intrinsic light is adapted to excite fluorescent light from the fluorescent proteins, and wherein the intrinsic light and the fluorescent light are diffuse. A method of optical tomography includes generating the excitation light with the apparent light source, wherein the intrinsic light and the fluorescent light are diffuse.

The invention discloses a yellow fluorescent dye, a method for synthesizing the same and application. The fluorescent dye has a structure shown as the general formula (I), wherein R1 and R2 are selected from chlorine, fluorine, bromine or hydrogen; and n is a positive integer between 3 and 7. The fluorescence emission spectrum of the yellow fluorescent dye is positioned in a yellow light region of visible spectrum; and the yellow fluorescent dye has high fluorescence quantum yield and good fluorescence labeling capacity, and can be used for protein labeling.

The invention discloses a yellow fluorescent powder and a preparation method thereof and a light emitting device prepared from the fluorescent powder. The fluorescent powder has the chemical formula of A[x]Z[y]Ce[3-x-y]E[m]D[5-m]O[12], wherein A is one or more elements of La, Y, Lu and Tb, Z is one or more elements of Zn, Mg, Ca, Sr and Ba, E is one or more elements of Al, In and Ga, D is one or two of Si and Ge, 0<x<3, 0<y<=0.2, x+y<3, 4.8<=m<5, and y+m=5. The yellow fluorescent powder has the advantages of low production cost, simple process and higher light emitting intensity.

The invention relates to a semiconductor LED fluorescent packaging structure. The semiconductor LED fluorescent packaging structure comprises a packaging substrate, wherein an LED element is attached to the surface of the packaging substrate and arranged in a transparent diffuse reflection layer on the packaging substrate, a first fluorescent layer is arranged on the outer surface of the transparent diffuse reflection layer, a second fluorescent layer is arranged on the outer surface of the first fluorescent layer, and a fluorescent protective layer is arranged on the outer surface of the second fluorescent layer. According to the semiconductor LED fluorescent packaging structure, yellow fluorescent powder and red fluorescent powder are arranged on the diffuse reflection layer, the problem of nonuniformity caused by mixed use of the yellow fluorescent powder and the red fluorescent powder is avoided, attenuation of the fluorescent powder is relieved, and the lighting effect of LEDs is improved. Meanwhile, total reflection in the packaging structure is reduced, and luminous efficiency is improved.

The invention discloses a packing technique for LED lamp beads; the technique comprises the following steps: A. LED chips are bonded on a bracket by insulation paste and are dried; B. gold threads are welded between electrodes and pins of the LED chips; C. red fluorescent powder, green fluorescent powder and yellow fluorescent powder are evenly mixed in part by weight: 20 to 25 parts of red fluorescent powder, 35 to 40 parts of green fluorescent powder and 40 to 45 parts of yellow fluorescent powder; D. 0.1 to 3 parts of rare earth raw material by weight are added in the mixed fluorescent powder materials of the step C and are evenly stirred; E. the mixed fluorescent powder materials prepared in the step D are coated on the LED chips to form fluorescent powder layers; and F. the glue injection and the formation are carried out for the LED lamp beads. According to the LED lamp beads produced by the packing technique disclosed by the invention, the light thereof can achieve the requirements of warm-color light.

A composition includes an artificial construct having (a) a reporter-containing portion chemically coupled to (b) a cleavage site. The cleavage site interacts with an investigational substance in a manner that cleaves the reporter-containing portion from a remainder of the construct. The cleaved portion is destroyed or otherwise degraded by the local environment, and presence of an investigational substance is evidenced by reduction in signal from the reporter. The investigational substance is preferably a Botulinum toxin (BoTN), and the cleavage sequence is all or part of a SNARE protein. The cleavable reporter-containing portion is preferably Yellow Fluorescent Protein (YFP), Citrine, Venus, or a YPet protein.

The invention provides a transformation method utilizing a red fluorescent protein as a selection marker. The method comprises the following steps of: synthesizing a fluorescent protein (FP) gene and modifying according to the bias of a rice codon, wherein the modified FP gene is used as a selective marker gene of rice transformation and plant regeneration; driving the FP gene by a callus / seed coat-specific promoter, closely connecting with a target gene, and transferring to rice embryonic calli under the mediation of Agrobacterium tumefaciens; screening for three times by a fluorescence microscope in 30 days after coculture; and further verifying an obtained transgenic seedling by utilizing a polymerase chain reaction (PCR) and Southern hybridization. The result shows that the positive rate of the transgenic seedling is up to 80 percent, which proves that FP serving as the selection marker of plant transformation is completely feasible. By the method, the potential hazard of the traditional selection markers such as antibiotic and herbicide resistance genes and the like to environment and food safety is effectively reduced. The invention provides a novel safe plant genetic transformation screening system.

The invention discloses a fluorescent protein, a fusion protein, an isolated nucleic acid, a vector and application thereof, belonging to the technical field of biomedical optics and molecular imaging. The fluorescent protein disclosed in the invention further comprises the amino acid mutation site C158D compared with the conventional fluorescent protein mCrimson. The mutation site enables the fluorescence spectral properties of the fluorescent protein of the invention to be changed, and the fluorescent protein has substantial Stokes shift characteristics, is applicable to single-photon or multi-photon microscopic multicolor imaging, can effectively reduce crosstalk between the fluorescence spectra of different fluorescent proteins, enhances the sensitivity of imaging, ensures simultaneousfluorescent imaging of the fluorescent proteins with different spectra and reduces high requirements on optical devices.

The present invention provides variants of fluorescent proteins, which are improved with regard to their properties for use as reporter proteins and / or in analytics. In particular, variants of fluorescent proteins are provided, which fluoresce brighter, show improved quantum yield and / or have shifted excitation or emission spectra. The fluorescent proteins according to the invention comprise in their LOV domain besides the substitution of a cysteine with an amino acid that does not covalently bind FMN at least one further point mutation.

The invention discloses a preparation method of a tumor detection nanoprobe. The method comprises the following steps of: (1) constructing expression plasmids for expressing a ferritin heavy chain, expressing with colon bacillus, purifying ferritins and verifying and representing a cage structure with an electron microscope; (2) fusing a green fluorescent protein at an amino terminal of each ferritin and performing expression and purification to obtain a ferritin cage structure of which the surface expresses the fluorescent protein; (3) fusing a segment of tumor targeting small peptides RGD (Arginine-Glycine-Aspartate) at the amino terminal of a ferritin H chain of which the amino terminal is fused with the green fluorescent protein to prepare a ferritin cage structure of which the surface expresses the fluorescent protein and the tumor targeting peptides; and (4) synthesizing Fe3O5 nanoparticles in the cavity of the ferritin cage structure of which the surface expresses the fluorescent protein and the tumor targeting peptides simultaneously to obtain a multifunctional ferritin probe with tumor targeting, fluorescence and magnetism. In the nanoprobe, a tumor targeting function, a fluorescent imaging function and a magnetic resonance imaging function are fused integrally, so that synchronous detection in multiple modes can be realized.

Provided are four new fluorescent proteins. The proteins were derived from two wild-type fluorescent proteins: a red fluorescent protein (RFP) that was isolated from Actinodiscus or Discosoma sp. 1 and a green fluorescent protein (GFP) isolated from Montastraea cavernosa. Two mutant forms were generated from each wild-type protein. Each of the mutated forms has a higher fluorescence intensity than the respective wild-type form. The mutant forms of the fluorescent proteins allow for more sensitive detection of the fluorescence emitted by the proteins. Additionally, one of the mutant proteins is more resistant to photobleaching than its wild-type protein. The invention also encompasses isolated nucleic acids encoding the mutant forms of the wild-type RFP and GFP.

The biomedicine PTD4-GFP-Apoptin fusion protein for treating tumor has its synthesized PTD4 utilized to introduce Apoptin protein into cell, and its Apoptin further utilized to induce tumor apoptosis, so as to inhibit tumor growth. At the same time, green fluorescent protein (GFP) is utilized for observing the intracellular distribution of the fusion protein and showing the aggregation of the medicine in the target position, realizing the effective monitoring and timely regulation of the treating process. Experiment shows that the fusion protein can penetrate biomembrane to kill tumor cell effectively while generating no damage to health cell. The present invention is applied in treating tumor.