31 results about "Fluorescent tag" patented technology

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.

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 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 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.

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.

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

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 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.

The invention provides a recombinant coccidiosis vector for expressing VP2 protein and a fluorescent label and a detection method of the recombinant coccidiosis vector, and belongs to the technical field of genetic engineering. The invention also provides sgRNA (small guide ribonucleic acid) targeting the ETH00009555 gene, and the sgRNA can be used for carrying out accurate gene fixed-point insertion; a gene editing system is established by sgRNA and homologous recombinant fragments, so that target fragments such as chicken infectious bursal disease virus VP2 protein and fluorescent label protein can be accurately subjected to homologous recombination on the ETH00009555 gene, and a recombinant coccidiosis vector for expressing the VP2 protein and a fluorescent label is constructed. The recombinant coccidium vector DNA is subjected to PCR (Polymerase Chain Reaction) detection and sequencing, and the result shows that the VP2 gene and the fluorescent tag gene are successfully recombined with the ETH00009555 gene; and the recombinant coccidiosis vector has an obvious fluorescence signal under a fluorescence microscope, which indicates that the recombinant coccidiosis vector successfully expresses the VP2 protein and the fluorescence tag protein.

A method for extending a depth of field of a nucleic acid sequencer may comprise optimization steps which are repeated one or more times, in which a result of passing light through an objective lens and a mask is compared with an ideal result, and any discrepancy is used to update the mask. Such a mask may be incorporated into a nucleic acid sequencer which adds fluorescent tags to nucleic acid sites and then detect light emitted from the fluorescent tags, thereby extending the sequencer's depth of field.

A method for extending a depth of field of a nucleic acid sequencer may comprise optimization steps which are repeated one or more times, in which a result of passing light through an objective lens and a mask is compared with an ideal result, and any discrepancy is used to update the mask. Such a mask may be incorporated into a nucleic acid sequencer which adds fluorescent tags to nucleic acid sites and then detect light emitted from the fluorescent tags, thereby extending the sequencer's depth of field.

The invention provides an immune MoS < 2 > atTQD fluorescent label as well as a preparation method and application thereof, and relates to the technical field of lateral chromatographic detection. The immune MoS < 2 > (at) TQD fluorescent label is a two-dimensional molybdenum sulfide quantum dot-based nano material labeled by an anti-carboxyl fluorescein antibody; the nanometer material based on the two-dimensional molybdenum sulfide quantum dots sequentially comprises a molybdenum disulfide nanosheet, a first coating layer, a second coating layer and a third coating layer from inside to outside, wherein the first coating layer, the second coating layer and the third coating layer are PEI layers for adsorbing CdSe-ZnS quantum dots. Meanwhile, the invention further provides the immune MoS2 (at) TQD fluorescent label, and the immune MoS2 (at) TQD fluorescent label is the quantum dot nanometer material based on two-dimensional molybdenum sulfide, which is marked by adopting an anti-carboxyl fluorescein antibody. The label can be used as an excellent signal label of a nucleic acid on-site rapid detection system, shows more excellent colorimetric and fluorescence signals, and is better in stability.

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.

A composition for treating pain includes an antibody or antibody fragment that specifically binds to a receptor involved in pain. In some embodiments, the receptor is a P2X family receptor such as, for example, P2X4. In some embodiments, the antibody includes a detectable marker. In some of these embodiments, the detectable marker includes a fluorescent tag. The composition may be used to treat acute or chronic pain in a subject. Generally, the methods includes administering to the subject a composition that includes an antibody or antibody fragment that specifically binds to a receptor involved in pain. The non-opioid pain relief composition can reverse pain related mechanical, cold, anxiety-like, and depression-like behaviors in neuropathic pain and chemotoxic nerve injury models.

A microfluidic reaction chamber with a reaction chamber circuit includes a microfluidic reaction chamber to contain a reaction fluid for amplification of nucleic acids, and a reaction chamber circuit disposed within the microfluidic reaction chamber. The microfluidic reaction chamber includes a base wall, a top wall parallel to the base wall and defined in part by a transparent lid, a first side wall, and a second side wall. The reaction chamber circuit is disposed within the microfluidic reaction chamber, and includes a top surface, a bottom surface, a first side wall, and a second side wall. The reaction chamber circuit is in fluidic contact with the reaction fluid and includes a photodetector to detect a fluorescence signal from a labeled fluorescent tag in the reaction fluid.

The invention discloses a CBA method reagent for detecting a GFAP antibody through fusion expression of GFAP alpha and GFAP epsilon and a detection method, and belongs to the technical field of neural immunodiagnosis. The fusion protein sequentially comprises the following functional domains: secretion signal peptide, a GFAP alpha partial sequence, a GFAP epsilon partial sequence, a transmembrane sequence and a fluorescent label. The core of the invention lies in that a fusion cell is constructed by using partial sequences of GFAP alpha and GFAP epsilon, and co-expression is realized in a host cell through a co-transfection technology, so that the sensitivity and specificity of detecting a CFAP antibody in a CBA method (cellular immunofluorescence method) can be remarkably improved.

The present invention addresses the problem of providing: a nerve cell analysis method with which the localized amount of TDP-43 in nerve cells can be analyzed without labeling TDP-43 with a fluorescent tag or the like and without carrying out gene transfer; a kit for carrying out the nerve cell analysis method; and a screening method for a prophylactic and / or therapeutic agent for a neurodegenerative disease. The present invention provides a nerve cell analysis method involving: a staining step for immunofluorescent staining of nerve cells using an anti-TDP-43 antibody and an antibody that recognizes a stress granule marker; a cell region identification step for identifying the cytoplasm and nuclei of the nerve cells; and an analysis step for analyzing a TDP-43-derived fluorescence signal and a stress granule marker-derived fluorescence signal in the cytoplasm, and analyzing the localized amount of TDP-43 in the nerve cells on the basis of the presence or absence of colocalization of a granular TDP-43 signal and a granular stress granule marker signal in the cytoplasm.

The invention provides a method for detecting a low-arsenic accumulation genotype of wheat grains based on a KASP marker, and relates to the technical field of wheat molecular breeding. Arsenic is a metalloid element with extremely high toxicity, and can enter wheat grains through soil pollution to threaten food safety, so that cultivation of wheat varieties with low arsenic accumulation is of great significance. According to the method, specific KASP primers are designed aiming at six stable genetic SNP sites for controlling arsenic accumulation of wheat grains, and the specific KASP primers comprise two allele specific forward primers carrying FAM / HEX fluorescent labels and a universal reverse primer; wheat genome DNA is used as a template, a 10 [mu] L reaction system is adopted for PCR amplification, and the procedure comprises a pre-denaturation stage, a Touchdown amplification stage and a fluorescence collection stage; the genotype is judged through a fluorescence signal, and the dominant haplotype related to low-arsenic accumulation is determined. The method has the advantages of strong specificity, high accuracy, realization of rapid detection in the wheat seedling stage, suitableness for high-throughput screening, provision of an accurate basis for molecular marker-assisted breeding, and acceleration of the cultivation of low-arsenic accumulation wheat varieties.

The invention relates to the technical field of immunological detection of small molecules in agricultural products, in particular to a GO-based double-color film fluorescent label and application of the GO-based double-color film fluorescent label in immunochromatography detection of pesticides, discloses an advanced double-color gradient fluorescence immunochromatography (DCG-FIC) system, and develops a double-color fluorescent nano-label. The sensitivity and the specificity of small molecule pollutant detection are obviously improved. The DCG-FIC method provided by the invention is substantially improved in detection performance, the detection limit of imidacloprid (IMI) is 0.611 pg / mL, the detection limit of azoxystrobin (AZO) is 2.16 pg / mL, and the detection limit is increased by 46 times compared with that of a traditional gold nanoparticle method. According to the method, rapid detection is conducted for 15 min, the adding standard recovery rate ranges from 85.13% to 105.14%, the relative standard deviation is smaller than 8.45%, and a promising solution is provided for screening of multiple pesticide residues of commercial crops.

This invention relates to the field of biotechnology and provides a gene-modified mouse model for tracking migratory organisms, its construction method, and its applications. The invention achieves precise labeling of endogenous migratory organisms by fusing the key structural protein TSPAN4 of the migratory organism with the fluorescent tag RFP and the modification tag Avi-tag, and site-specifically integrating it into the H11 site of mice, thus obtaining a TSPAN4-overexpressing transgenic mouse model. By introducing an LSL structure, the fusion gene is kept in a state of expression shutdown, achieving temporal and spatial specific activation only after mating with tissue-specific Cre tool mice. Multidimensional validation has shown that this model can accurately track migratory organisms in placental trophoblast cells, providing a standardized tool for assessing disease progression under physiological and pathological conditions through differences in migratory organism trajectory, inclusion packaging, and assembly, and can be directly applied to mechanistic studies and drug screening for diseases such as adverse pregnancy outcomes.

The application discloses a Ustilaginoidea virens dominant strain HEB07 and construction of a Ustilaginoidea virens pathogenic bacterium and millet interaction space-time atlas, relates to the technical field of biology, and the strain is screened and verified, the strain has the infection breadth of different millet varieties and the infection pathogenic sensitivity and survival rate of any millet variety, and has high affinity of recombination transformation of a GFP fluorescent label, so as to constitute a gene recombination strain using a green fluorescent protein GFP and explore the Ustilaginoidea virens pathogenic bacterium and millet interaction dominant strain. The application can provide important technical support for cloning research of millet disease-resistant genes, and provide strong scientific basis and technical means for cultivating millet varieties with high resistance to Ustilaginoidea virens.

The application discloses a recombinant protein for detecting paraneoplastic Yo antibody by CBA method and application, and belongs to the technical field of biological medicine and engineering. The amino acid sequence of the recombinant protein comprises, in sequence, a secretion signal peptide, a CDR2 protein partial sequence, a CDR2L protein partial sequence, a transmembrane region and a fluorescent label. The secretion signal peptide is a human albumin signal peptide ALB. The transmembrane region is CD8a hinge. The fluorescent label is mCherry. The application constructs a novel recombinant protein which can be stably overexpressed on the cell membrane of eukaryotic cells by intercepting specific partial sequences of CDR2 protein and CDR2L protein, and redesigning and fusing a fluorescent label by using a secretion signal peptide, a transmembrane sequence and a connecting peptide. The recombinant protein retains the core antigen regions of the two, can effectively overcome the insufficiency in sensitivity and specificity, can significantly improve the detection rate of paraneoplastic Yo antibody when CBA method is used for detection, and can reduce the false positive rate, so that the requirement of clinical detection can be better met.

Fluorophore-containing compounds for use as fluorescent tags for the detection or analysis of carbohydrates are provided. Also described are linkers for the preparation of the fluorescent compounds, methods of preparing and using the fluorescent compounds, and carbohydrate conjugates formed from the compounds.

The invention belongs to the field of food packaging materials, and provides a preparation method of a fluorescent label for specific detection of putrescine, tyramine and cadaverine, and the preparation method comprises the following steps: dispersing rhodamine B into an ethanol solution to prepare an indicator I; dissolving polymethylacrylic acid and hydroxytetraphenyl ethylene in tetrahydrofuran, and preserving heat to obtain an indicator II; dissolving zein in an ethanol solution, heating and stirring, cooling, and adding a plasticizer to obtain a loaded matrix solution; uniformly mixing the indicator I and the indicator II, slowly adding the loaded matrix solution to obtain a film-forming solution, degassing, casting, spreading, drying, and balancing temperature and humidity to obtain the fluorescent label. The fluorescent label prepared by the invention is wide in concentration response range to putrescine, tyramine and cadaverine solutions, low in detection limit, sensitive in response and high in color discrimination, and can accurately indicate the freshness of fresh chicken.

The application discloses a CBA method reagent for detecting GFAP antibody by fusing expression of GFAP alpha and GFAP epsilon and a detection method, and belongs to the technical field of neuroimmunological diagnosis. The reagent comprises a host cell which is recombined to express a fusion protein. The fusion protein comprises the following functional domains in sequence: a secretion signal peptide, a GFAP alpha partial sequence, a GFAP epsilon partial sequence, a transmembrane sequence and a fluorescent label. The core of the application is that the fusion cell is constructed by using the partial sequences of GFAP alpha and GFAP epsilon, and the co-expression is realized in the host cell by using the co-transfection technology. The sensitivity and the specificity of the detection of the CFAP antibody in the CBA method (cellular immunofluorescence method) can be significantly improved.

Disclosed probes comprise metal nanoparticle cores associated with magnetic particles that allow probes associated with targets to be concentrated by an applied magnetic field to increase detection sensitivity and provide sufficient spacing between concentrated probes to avoid signal quenching. The probe may comprise at least one recognition receptor, and may further comprise at least one reporter molecule, such as a fluorescent tag, a Raman reporter, or combinations thereof. Concentrating probe-target composites substantially enhances a sensing signal, such as from 5 to 10 times, compared to detection without concentrating the probes. The method may be used to detect, for example, interleukins at concentrations at least as low as 25 pg / ml in sputum or blood from a subject for early and precise profiling of viral infections, such as SARS-CoV-2 infections.