58 results about "Fluorescent tag" patented technology

The invention discloses a recombinant protein and a detection reagent for detecting encephalitis of an anti-GABABR antibody, and belongs to the technical field of neuroimmunology diagnos.A GABABR1 partial sequence, a GABABR2 partial sequence and a transmembrane sequence (and a fluorescent label) are adopted to construct a cell strain for stably and highly expressing the recombinant protein by transfecting CHO cells or HEK293 cells, and the recombinant protein and the detection reagent are used for detecting the encephalitis of the anti-GABABR antibody. The sensitivity and the specificity of the kit in anti-GABABR antibody encephalitis detection can be remarkably improved, the aim of detecting the antibody by using a fixed CBA method or a living cell CBA method can be achieved, and the kit is suitable for auxiliary diagnosis of autoimmune encephalitis of cerebrospinal fluid / serum.

The invention discloses a recombinant protein for detecting a para-tumor Yo antibody through a CBA method and application, and belongs to the technical field of biomedical engineering.The amino acid sequence of the recombinant protein sequentially comprises a secretory signal peptide, a CDR2 protein partial sequence, a CDR2L protein partial sequence, a transmembrane region and a fluorescent label, and the secretory signal peptide is human albumin signal peptide ALB; the transmembrane region is a CD8a hinge; and the fluorescent label is mCherry. A novel recombinant protein which can be stably over-expressed on a cell membrane of an eukaryotic cell is constructed by intercepting specific partial sequences of CDR2 protein and CDR2L protein and redesigning and fusing a fluorescent label by using a secretory signal peptide, a transmembrane sequence and a connecting peptide, and the recombinant protein retains respective core antigen regions of the CDR2 protein and the CDR2L protein, so that the specific partial sequences of the CDR2 protein and the CDR2L protein can be stably over-expressed on the cell membrane of the eukaryotic cell. The kit can effectively overcome the defects in the aspects of sensitivity and specificity, and when a CBA method is adopted for detection, the detection rate of the para-tumor Yo antibody can be remarkably increased, and the false positive rate is reduced, so that the requirements of clinical detection are better met.

This disclosure provides a method of tagging binders in electrodes to allow for easy visualization. In particular, environmentally friendly organic molecules are used as organic fluorescent tags. The organic fluorescent tag will react with alkene moieties within the binder through cycloadditions to introduce a fluorescent tag, which is easily visualized to allow visualisation of binder distribution within the electrode.

The invention discloses a method for screening bacterial strains with an antibacterial function, which comprises the following steps: to-be-screened bacteria and indicator bacteria are screened in a microfluidic system, the indicator bacteria have a fluorescent label so as to obtain target bacteria, and the target bacteria have the activity of inhibiting the indicator bacteria. According to the method, the antagonistic activity of the strains is associated with the fluorescence signal intensity, the functional strains with the antibacterial function are screened in a high-throughput and high-precision mode, the screening throughput reaches 106-108 strains every day, and therefore the screening efficiency and the result stability are remarkably improved.

The invention provides a KASP molecular marker related to mesosulfuron resistance of wheat and application of the KASP molecular marker, and belongs to the technical field of crop molecular genetic breeding. Through correlation analysis, it is found that the KASP molecular marker is remarkably correlated with mesosulfuron-methyl resistance, T / C polymorphism exists, and the mesosulfuron-methyl resistance of CC homozygous wheat is remarkably higher than that of TT homozygous wheat. On the basis, a KASP labeling reagent or kit containing a specific primer group is developed, the primer group comprises two allele specific upstream primers and a universal downstream primer, and the 5'ends of the upstream primers are connected with FAM and HEX fluorescent labels respectively. By optimizing a PCR reaction system and a landing procedure, high-throughput genotyping can be realized. The mesosulfuron-methyl-resistant wheat variety can be rapidly screened by applying the KASP molecular marker, a technical support is provided for herbicide resistance molecular breeding, the breeding efficiency of the resistant variety is remarkably improved, and the KASP molecular marker has important significance for guaranteeing safe production of wheat.

The present invention discloses a fluorescent probe and a fluorescent label for real-time non-destructive visualization monitoring of the freshness of meat food, and their preparation methods and applications. The structural formula of the fluorescent probe HBT-Bz is simple and easy to synthesize. Using the HBT fluorophore with ESIPT properties and AIE effect as the parent body and benzoxazole ester group as the recognition group for amine compounds, a fluorescence turn-on probe is designed and synthesized. The fluorescent probe HBT-Bz responds quickly and sensitively to amine compounds, with a detection limit as low as 5.1 ppm, and can be used to fabricate a solid fluorescence sensor for detecting volatile amine compounds. The synthetic route of the fluorescent probe involved in the present invention is simple, can recognize a variety of volatile amines, has a low detection limit, and the preparation process of the fluorescent label loaded with the fluorescent probe is simple. When in use, no pretreatment of the food is required. Placing the fluorescent label in the meat food packaging can realize real-time non-destructive visualization monitoring of the food freshness, and the results are accurate and reliable.

The application discloses a fluorescent label for nondestructive visual monitoring of freshness of fish and meat food and a preparation method thereof. The fluorescent label is characterized in that a key fluorescent substance JDCN specifically interacts with volatile biogenic amines generated in the meat spoilage process, color and fluorescence double optical signal changes occur, the biogenic amine content is judged according to the visual optical characteristics, and thus the freshness of the fish and meat food is monitored. The application has the characteristics of simple preparation, low cost, high sensitivity, high selectivity, fast visual response, and the like, can in-situ rapidly detect the volatile biogenic amines generated in the spoilage of the fish and meat food, provides a simple, convenient and effective means for the rapid visual detection of the freshness of the fish and meat food, is expected to become a preferred method for rapid detection of the content of the volatile biogenic amines in the fish and meat food, provides a powerful technical guarantee for self-monitoring of food manufacturers and market supervision, has great application prospect, and is of great popularization significance.

This invention relates to the field of plant molecular breeding technology, specifically to a KASP molecular marker PH5-KASP based on the SNP site of the maize Zm00001eb259660 gene and its application. It provides a KASP molecular marker targeting the SNP site at 226029364 bp on maize chromosome 5, which is a functional missense mutation c.485C>T in the coding region of the Zm00001eb259660 gene. The marker consists of two allele-specific forward primers carrying different fluorescent tags and one universal reverse primer, stably distinguishing between C:C and T:T homozygous genotypes, where C:C corresponds to higher plant height and T:T corresponds to lower plant height. This invention offers high marker genotyping accuracy, simple and low-cost detection, and is suitable for large-scale breeding sample detection. It can be used for early identification of maize plant height traits, germplasm screening, and assisted breeding, significantly improving the efficiency of ideal maize plant architecture improvement.

The present disclosure relates to a method of identifying two or more characteristics of bacterium in a testing sample comprising use of fluorescent labels and hybridization chain reaction.

The invention relates to the field of bioengineering, in particular to a recombinant human herpes simplex virus as well as a construction method and application thereof. The recombinant human herpes simplex virus lacks US1, US2, US3, US4 and US5 virulence genes at the same time. The construction method comprises the following steps: constructing a donor plasmid, and constructing a cleavage plasmid; transfecting cells with the cleavage plasmids and the donor plasmids in proportion; and after the transfected cells are incubated, infecting with an HSV-1 virus, purifying, and verifying to obtain the recombinant virus. The donor plasmid constructed by the invention retains non-coding regions among US1, US2, US3, US4 and US5 genes, also retains initiation codons and termination codons of the US1, US2, US3, US4 and US5 genes, is added with a fluorescent tag sequence, and can knock out five virulence genes of HSV-1US1-US2-US3-US4-US5 at the same time. The neutralizing antibody titer result of the recombinant human herpes simplex virus is the same as that of a wild type, and a foundation is laid for vaccine preparation.

The invention belongs to the technical field of animal reproductive biology and cell engineering, and particularly relates to a fluorescence labeling expression system for visualizing an oocyte nucleus maturation process and application of the fluorescence labeling expression system. The fluorescence labeling expression system comprises an eukaryotic expression vector pVenus-LMNB1; the eukaryotic expression vector pVenus-LMNB1 comprises a nucleotide sequence for coding the fusion protein Venus-LMNB1. According to the invention, a nuclear fiber layer protein stably expressed on a nuclear membrane is selected as a research object, a eukaryotic expression vector pVenus-LMNB1 is constructed by using a molecular cloning technology, an LMNB1 gene containing a green fluorescent label Venus is overexpressed in a porcine oocyte by using a microinjection technology, fluorescent protein labeling of a structural protein on the nuclear membrane is utilized, and the expression vector pVenus-LMNB1 is constructed. The expression positioning condition of the LMNB1 in the maturation process of the porcine oocytes is observed, and a powerful tool is provided for development of in-vitro culture related researches of the porcine oocytes.

The present disclosure is directed to a method for screening a signal peptide for efficient expression and secretion of a heterologous polypeptide in mammalian cells, the method comprising the steps of: providing a pool of viral expression vectors encoding a polypeptide of interest with various candidate signal peptides, wherein each viral expression vector of said pool comprises at least a polynucleotide encoding a fusion protein, said polynucleotide comprising: i) a promoter, ii) a sequence encoding a signal peptide, iii) a sequence encoding the polypeptide of interest, iv) optionally a sequence encoding an epitope tag, and v) a sequence encoding a C-terminal signal peptide for glycosylphosphatidylinositol, GPI, attachment or a sequence encoding a transmembrane domain; transforming host cells with said pool of viral vectors so that each host cell is preferably transformed on average by only one viral vector from said pool; expressing said fusion protein in said host cells in order to produce fusion proteins which are GPI-anchored to the host cell surface or alternatively which are anchored to the host cell surface by a transmembrane domain of said fusion protein; contacting said host cells with a first binding reagent, preferably an antibody, specifically binding to said polypeptide of interest or alternatively if said epitope tag is present in the fusion protein with a first binding reagent specifically binding to said epitope tag, wherein said binding reagent is optionally labelled with a fluorescent label or other means of detection; dividing the transformed host cells into at least two groups based on the fluorescence characteristics of each host cell; and performing next generation sequencing to the group of host cells showing the most efficient fusion protein expression in order to identify an optimal signal peptide for the polypeptide of interest in the host cell. The present disclosure is also directed to a vector, a host cell or a DNA library for use in said method.

This invention provides a KASP molecular marker associated with wheat resistance to mesosulfuron-methyl and its application, belonging to the field of crop molecular genetics and breeding technology. Association analysis revealed a significant association between this KASP molecular marker and the mesosulfuron-methyl resistance trait, exhibiting T / C polymorphism. Specifically, CC homozygous wheat showed significantly higher resistance to mesosulfuron-methyl than TT homozygous wheat. Based on this, a KASP marker reagent or kit containing a specific primer set was developed. The primer set includes two allele-specific upstream primers and one universal downstream primer, with FAM and HEX fluorescent tags attached to the 5' ends of the upstream primers, respectively. High-throughput genotyping can be achieved by optimizing the PCR reaction system and landing procedure. The application of this KASP molecular marker can rapidly screen wheat varieties resistant to mesosulfuron-methyl, providing technical support for herbicide-resistant molecular breeding, significantly improving the efficiency of resistant variety selection, and playing a vital role in ensuring safe wheat production.

The invention relates to the technical field of drug research and development, and discloses a screening model for high-throughput rapid screening of PROTAC protein degradation drugs. According to the model, a 96-well plate for cell culture is selected, the intensity of green fluorescence carried by an exogenous target protein is detected through a microplate reader, and the protein degradation activity of the PROTAC compound is rapidly screened. When the model is used for detecting the degradation activity of a PROTAC compound taking FGFR4 as a target spot, the method comprises the following steps: (1) inserting an FGFR4 gene into a Turbo GFP-Vector plasmid with a green fluorescent label; (2) instantaneously transfecting HEK293T cells for 24 hours; (3) adding a PROTAC compound to treat the cells for 24 hours; and (4) detecting a fluorescence value at the wave band of Ex482nm / Em502nm by a microplate reader so as to judge the activity of the PROATC compound for specifically degrading the expression of the exogenous FGFR4 protein. The 96-well plate for cell culture adopted by the model can achieve high-throughput live cell screening, and the exogenous target protein is expressed by using a transient transfection plasmid method, so that the method is more time-saving, simple, convenient and economic, and has strong repeatability.

The present invention provides a phenothiazine derivative, a synthesis method thereof, and its application in food freshness detection. The present invention designs and synthesizes a fluorescent molecule based on a phenothiazine derivative with protonation / deprotonation behavior. Using the protonated phenothiazine derivative as the detection target, and 1,4-butanediamine and 1,5-pentanediamine produced during food spoilage as the target, the fluorescent on-state behavior enables accurate and rapid detection of these targets. The target is then loaded onto a test strip to prepare a fluorescent label, enabling freshness detection of actual food samples at room temperature using the solution / test strip. Furthermore, for further convenience and practicality, an intelligent detection method has been constructed.

The invention provides a phenothiazine derivative as well as a synthesis method and application thereof in food freshness detection. The invention designs and synthesizes a fluorescent molecule based on a phenothiazine derivative with proton / deprotonation behavior, the protonated phenothiazine derivative is taken as a detection object, 1, 4-butanediamine and 1, 5-pentamethylene diamine generated in a food spoilage process are taken as detected objects, accurate and rapid detection is realized through a fluorescence opening behavior, and the detection sensitivity is high. And then the fluorescent label is prepared by loading the fluorescent label on a test strip, so that the freshness of an actual food sample is detected by the solution / test paper at normal temperature. In addition, in order to be convenient and practical, an intelligent detection method is constructed.

The present invention belongs to the field of biomedicine, and relates to an endogenous nanoprobe, a preparation method and an application thereof. The nanoprobe is an outer membrane vesicle carrying a non-oxygen-dependent controllable and adjustable fluorescent label, has a natural bacterial outer membrane structure, and can visually identify, track or inhibit targets in an aerobic or anaerobic environment. The size of the nanoprobe is 40-100nm. The endogenous nanoprobe has strong specificity and high efficiency, and its fluorescence has the flexibility of on-demand switching and reversible switching of emission bands, which can be used for intelligent and dual-color imaging, thereby significantly improving the sensitivity and accuracy of detection, and is not harsh on the required detection equipment and sites, and has low cost, and has important research significance and application value. The application of the endogenous nanoprobe includes: the use of the nanoprobe in the preparation of drugs for preventing, slowing down or treating intestinal diseases; a biosensor system comprising at least the nanoprobe of the present invention; and a bioimaging system comprising at least the nanoprobe of the present invention.

The present invention relates to substituted-diazenylanilines of the formula I and their nucleotide conjugates, complexes, salts which may be used potentially as fluorescent quenchers in chemical and biological sciences such as cell imaging applications, diagnostics, fluorescent and non-fluorescent tags, pharmaceuticals and other useful applications, and a process of preparing said new compounds. More particularly, the present invention relates to 2,2′-((4-((2,5-disubstituted-4-((4-nitrophenyl)diazenyl)phenyl)diazenyl)-2 / 3-substituted-phenyl)azanediyl)dialkanol, processes for preparing said compounds and their use as fluorescent quenchers in cell imaging applications, diagnostics, fluorescent and non-fluorescent tags, pharmaceuticals and other useful applications.

Disclosed are fluorescent tags comprising a cannabinoid fluorophore and compositions comprising the fluorescent tags.

The invention discloses an intercellular interaction quantitative detection method based on a monolayer cell adsorption experiment, which comprises the following steps: constructing an experiment system conforming to a Langmuir model, and respectively taking cells transfected with different fluorescent label interaction proteins as monolayer recipient cells and ligand cells, after incubation, observing under a fluorescence microscope, counting the number of adsorbed cells, and fitting a corresponding formula to obtain a dissociation constant Kd of the interaction between the cells, so as to realize quantitative evaluation on the affinity. The method provides a new visual angle and tool for studying the interaction between cells and understanding the function exertion of the cells and the interaction of membrane proteins on the surfaces of the cells, and solves the problems that the interaction between the cells cannot be effectively and quantitatively evaluated due to the technical limitation of high requirements on equipment and data, and the membrane proteins are low in content and insoluble in water; and a conventional protein purification method is difficult to obtain a high-purity and high-activity membrane protein sample and is difficult to accurately describe the binding affinity between membrane proteins.

The present application relates to dyes containing a 4,5-substituted naphthalimide core and their uses as fluorescent labels. These dyes may be used as fluorescent labels for nucleotides in nucleic acid sequencing applications.

The purpose of the present invention is to provide: a water-insoluble pigment composition in which a natural pigment and a dehydration condensation compound of a fatty acid and an amino acid are compounded; and a food, a cosmetic, a lipstick cosmetic, an eye cosmetic, a nail cosmetic, a base makeup cosmetic, a hair cosmetic, a pharmaceutical, an agrochemical coating material, a printing marker, stationery, a writing instrument, a printing ink, an inkjet ink, a metal ink, a coating material, a plastic coloring agent, a color toner, a fluorescent labeling agent, a fluorescent probe, or a chemical sensor containing the water-insoluble pigment composition.　The present invention was completed when it was discovered that compounding a natural pigment and a dehydration condensation compound of a fatty acid and an amino acid yields a water-insoluble pigment composition.

The invention relates to novel caging-group-free photoactivatable fluorescent dyes having the structural formula I:as well as to the corresponding photoactivated fluorescent dyes having the structural formula II:The invention further relates to the use of the photoactivatable compounds as such or after photoactivation, in particular as fluorescent tags, analytical reagents and labels in optical microscopy, imaging techniques, protein tracking, nucleic acid labeling, glycan analysis, capillary electrophoresis, flow cytometry or as a component of biosensors, or as analytical tools or reporters in microfluidic devices or nanofluidic circuitry.

The application relates to the field of agricultural biotechnology, and particularly discloses a KASP molecular marker for identifying the resistance of soybeans to phytophthora root rot and application. The KASP molecular marker primer sequence is shown in the upstream primer SEQ ID No. 1 (connected with a FAM fluorescent tag), the upstream primer SEQ ID No. 2 (connected with a VIC fluorescent tag) and the universal downstream primer SEQ ID No. 3. The KASP molecular marker can realize efficient, accurate and low-cost identification and genotyping of the resistance of soybeans to phytophthora root rot, and the effective identification and detection reaches 98.55%, and the accuracy reaches 84.80%.

The invention discloses a construction method and application of an acetylcholin esterase inhibitor high-throughput screening cell line. The invention belongs to the technical field of biology, and particularly relates to a construction method and application of an acetylcholin esterase inhibitor high-throughput screening cell line. The recombinant cell constructed by the invention contains a fluorescent label expression module and an acetylcholin esterase gene expression cassette, wherein the fluorescent label expression module contains a cpGFP-inserted muscarinic-like receptor (M3R) encoding gene and a red fluorescent protein mcherry encoding gene carrying a CAAX membrane positioning sequence. When the recombinant cell is used for screening the acetylcholin esterase inhibitor, acetylcholin esterase does not need to be additionally added, the reaction step of acetylcholin esterase hydrolysate and auxiliaries is reduced, the use of toxic and unstable DTNB and other reagents is avoided, the system is simpler, and the time cost and the economic cost are lower.

The disclosure provides cartridges that are pre-loaded with reagents for performing antimicrobial susceptibility testing (AST) and FISH testing. Cartridges of the disclosure include various incubation wells loaded with different antimicrobial agents for differential growth analysis. Imaging wells with species-specific microbial probes, fluorescent tags, magnetic particles and dye-cushion layers allow for tagging and imaging of target microbes for differential growth analysis.

The invention relates to a method for detecting and / or screening a target protein expression cell line based on fluorescence indication, and aims to realize rapid detection of specific protein conformation in chassis cells and efficiently screen chassis cells with different functional states, activities and target protein expression levels. The essential difference between the technology and a conventional protein fusion fluorescent label for indicating protein is that the conformation change of the protein is reflected through a cross-linking agent, and the functional state corresponding to the conformation of the protein is reflected based on the change of the fluorescence color of a cross-linking agent skeleton under different protein conformations. The technology provides a convenient, efficient and non-invasive method for the field of biological research, and can be widely applied to the fields of cell activity research, drug screening, protein function research and the like.

The present application relates to dyes containing a 4,5-substituted naphthalimide core and their uses as fluorescent labels. These dyes may be used as fluorescent labels for nucleotides in nucleic acid sequencing applications.

This disclosure provides a slurry for forming an electrode comprising a tagged binder. The tagged binder comprises a polymer which is bound to an organic fluorescent tag, which introduces a fluorescent moiety to the binder. During or after battery fabrication, said fluorescent moiety allows for easy visualization of binder migration within the electrode.

The present application relates to a method for detecting and / or screening a target protein expression cell line based on fluorescence indication, aiming to realize rapid detection of specific protein conformation in a chassis cell, and at the same time, to efficiently screen chassis cells with different functional states, activities and target protein expression levels. The essence of the present technology is different from the conventional protein fusion fluorescent tag used to indicate the protein. The present method first reflects the conformational change of the protein through a cross-linking agent, and reflects the functional state corresponding to the conformation of the protein based on the change of the fluorescence color of the cross-linking agent skeleton under different protein conformations. This technology provides a convenient, efficient and non-invasive method for the field of biological research, which can be widely used in the fields of cell activity research, drug screening and protein function research.