83 results about "Glyceraldehyde phosphate" patented technology

The invention provides a reagent for detecting a Notch signal path as well as a PCR (Polymerase Chain Reaction) detecting method and application thereof. The detecting reagent comprises PCR reaction primers for detecting an ADAM10 gene, an ADAM17 gene, an AES gene, a CBL gene, a CCND1gene, a CD44 gene, and the like, and primers of corresponding internal-reference regulatory genes GAPDH (Reduced Glyceraldehyde-Phosphate Dehydrogenase), ACTB (Beta-Actin) and B2M (Beta-2-Microglobulin). The invention provides the reagent for detecting core molecule changes of the NOTCH signal channel; by virtue of the detecting reagent, genes associated with the NOTCH channel can be quickly detected on transcriptional level by virtue of real-time fluorescent quantitative PCR; on this basis, core molecules and a mechanism of action of a tumor NOTCH signal channel are analyzed and researched, so that an NOTCH regulatory channel of tumor-associated medicaments is quickly and accurately found, and therefore, a powerful tool is provided for screening anti-tumor medicaments, discussing the mechanism of a novel targeted medicament, and the like.

The invention discloses a preparation method of a periodic wuchereria malayi compound multivalent DNA (deoxyribonucleic acid) vaccine. The preparation method comprises the following steps of: designing a primer, amplifying BmGAPDH (reduced glyceraldehyde-phosphate dehydrogenase) and BmCPI genes by PCR (polymerase chain reaction), purifying segments of BmGAPDH and BmCPI genes, connecting and converting a T carrier, screening and identifying an I positive recombinant clone, building and identifying an I recombinant expression vector, extracting and purifying a mass of recombinant plasmid and the like. The preparation method is convenient in preparation and good in effect. The periodic wuchereria malayi which is popular in China is taken as a research object, and the periodic wuchereria malayi cysteine proteinase inhibitor/3-phosphoglyceraldehyde dehydrogenase eukaryotic expression recombinant plasmid is built. The test result proves that the compound multivalent DNA vaccine can induce the mouse to generate the specific immune response reaction, so that a satisfied effect can be obtained, and the periodic wuchereria malayi compound multivalent DNA vaccine can be successfully prepared.

The invention relates to an ammonia diagnosing/measuring reagent box which utilizes the technique of the enzyme contrasting color method and the enzyme jointing method, and belongs to the technical field of the medicine/food/environment test. The main components of the reagent box of the invention mainly include cushion liquid, coenzyme, glutamic acid, adentosine triphosphoric acid, glyceraldehyde-3-phosphate, glutamine synthetase, glyceraldehyde-3-phosphate dehydrogenase and stabilizing agent; the reagent box generates a series of enzymic reactions through the mixing of the samples and the reagent at a certain cubage rate, and then the reactants are positioned under an ultraviolet/visible light analyzer which tests the reducing speed of the absorbency at the main wave length of 340nm, thereby measuring the concentration size of the enzyme. The invention can absolutely get needed measuring result through the ultraviolet/visible light analyzer.

According to the present invention, an inorganic phosphoric acid is detected by a method which includes: subjecting a sample to a measurement system containing glyceraldehyde-3-phosphate, oxidized nicotinamide adenine dinucleotide or oxidized nicotinamide adenine dinucleotide phosphate, glyceraldehyde phosphate dehydrogenase, and an electron mediator; and measuring a current value in the measurement system. In the method, a pyrophosphate is quantitatively measured with high sensitivity and at a high speed through converting the pyrophosphate in a sample into an inorganic phosphoric acid. Such a measurement of a pyrophosphate allows for quantitative determination of the pyrophosphate which is produced concurrent with the extension of a DNA, thereby enabling the detection of the presence of a targeted nucleic acid, and typing of a base in a SNP site of a targeted DNA.

The invention discloses a selection method of reference genes in quantitative real-time PCR analysis of jerusalem artichoke, and relates to the field of quantitative PCR. The method comprises the following steps: taking different tissues (radicles, immature stems, leaves, stem blocks and petals) of the jerusalem artichoke as materials, and carrying out expression analysis of the reference genes of18S ribosomal RNA gene (18S rRNA), transcription elongation factor gene (Ef-1a), actin gene (Actin and beta-actin), 3-glyceraldehyde phosphate dehydrogenase (GAPDH), 25S ribosomal RNA gene (25S rRNA)and poly-ubiquitin enzyme gene (UBQ)7 by using a q PCR technology; carrying out statistical assessment on the obtained data and analyzing expression change of all housekeeping genes by utilizing GeNorm and NormFinder software, so as to screen out relatively-stable genes as the reference genes of the jerusalem artichoke, which are used for studying the gene dosage changes of the jerusalem artichoke.

The invention provides an L-lactic acid catalysis reaction system and a preparation method of L-lactic acid. The L-lactic acid catalysis reaction system contains glucose dehydrogenase, dihydroxyacid anhydrase, 2-keto-3-deoxygluconate, glyceraldehyde phosphate dehydrogenase and L-lactic dehydrogenase. The L-lactic acid can be obtained without adding ATP (Adenosine Triphosphate) by performing a cascade enzymatic catalytic reaction in a reactor by utilizing five enzymes of the reaction system and taking glucose as a raw material.

The invention provides a method for detecting salidroside synthetase gene expression activity by using dual reference genes. Specifically, a primer which is obtained through design and screening is adopted to amplify to obtain glyceraldehydes phosphate dehydrogenase and phytochelatin synthetase genes, and correct and detect the gene expression activity of three key enzymes, namely, tyrosine decarboxylase, tyrosine transaminase and phenylalanine ammonialyase of synthesized salidroside in rhodiola crenulata tissue culture seedlings by using the gene as the dual reference genes. The method comprises the following steps: extracting total RNA of a rhodiola crenulata plant; synthesizing a cDNA first chain; and by using a fluorescent quantitative PCR technique, amplifying by using a given primer so as to obtain the dual reference genes of GAPDH and PCS, and correcting and detecting the expression activity of synthesized salidroside key enzyme genes TyDC, TAT and PAL in the rhodiola crenulata tissue culture seedlings. The method has the advantages of rapidness, accuracy, practicability, stability and the like in aspects such as production quality control and product activity detection on rhodiola crenulata tissue culture seedlings, cell suspension culture, hairy root culture and the like.

The invention discloses a shikimic acid synthesis method. According to the method, fructose 6-phosphate and glyceraldehydes-3-phosphate ester serve as raw materials, and shikimic acid is generated under the catalytic function of various enzymes. The generation technology is simple in step, reagents are safe, byproducts are few in the reaction process, economic benefits are improved, the yield of the obtained shikimic acid is 50-86%, and the purity is 98.1-99.5%.

The invention relates to a zinc diagnosis/determination reagent kit which adopts the techniques of enzymatic colorimetric method and enzyme couple reaction, belonging to the technical field of medicine/food/environment test and determination. The invention also relates to the method principle of determining the zinc concentration, and the reagent composition and components. The component of the reagent kit mainly comprises buffer solution, coenzyme, phosphate monoester, glyceraldehyde-3-phosphate, alkaline phosphatase, glyceraldehyde-3-phosphate dehydrogenase and stabilizer. The method for determining the zinc concentration is as follow: mixing the sample and the reagent according to a certain volume ratio to perform a series of enzymatic reactions; placing the reactant under an ultraviolet / visible light analyzer to detect the ascending speed of the absorbency at the main wavelength of 340 nm; finally determining the zinc concentration. The invention has the advantages that the required determining result is obtained only with the ultraviolet / visible light analyzer.

The invention relates to a kit for identifying the genetype of a silky fowl skin color by detecting EDN3 (endothelin 3) gene copy number difference. The kit comprises a reference gene GAPDH (reduced glyceraldehyde-phosphate dehydrogenase) amplification primer (1), a target gene EDN3 amplification primer (2), calibration sample DNA (deoxyribonucleic acid) (3), fluorescent quantitative PCR (Polymerase Chain Reaction) reaction liquid (4) and double distilled water (5). The kit is characterized in that real-time fluorescent quantitative PCR reaction is carried out on a target gene EDN3 and a reference gene GAPDH by using the DNA of a tested sample as a template, Ct values are respectively recorded by using the calibration sample DNA as a reference, the EDN3 gene copy numbers of the tested sample are obtained by calculation, and further the Fm genetype of the silky fowl skin color is judged according to the EDN3 gene copy number difference. The operation of the kit does not need the establishment of a standard curve; the kit is simple, convenient, fast, good in repeatability and high in detection efficiency; and through chicken breed test-crossing testing, the method can accurately reflect the Fm homozygosis and heterozygosis of the skin color of the individual silky fowl. The detection method is conductive to acceleration of screening of homozygosity of the silky fowl skin color, guides target breeding of high-quality silky fowl new variety lines and provides reliable germplasm guarantee for follow-up commercialization mating hybridization.

The invention provides a kit for detecting the expression level of ERCC1 (excision repair cross complementation 1). By adopting a fluorescent quantitative PCR (polymerase chain reaction) platform, and taking RNA (ribonucleic acid) reserved by culture cells as external calibration RNA and GapDH (reduced glyceraldehydes-phosphate dehydrogenase) as data of internal contrast gene normalization processing experiments, relative quantitative detection is carried out on the level of mRNA (messenger ribose nucleic acid) of ERCC1 of a sample tissue through a TaqMan probe method. The kit can be used for detecting the expression quantity of the ERCC1 at a level of mRNA and can reflect the gene expression level more truly; the detection data of the fluorescent quantitative PCR method is output by a machine, thus avoiding subjective identification of people and being more objective; the cultured cells are taken as the external RNA calibrator, and the whole process and the fluorescent quantitative PCR experiment are synchronous, so that the errors caused by the experiment process is reduced, the determination of the clinical cut-off value is realized, furthermore, the fluorescent quantitative PCR method for detecting the level of mRNA of ERCC1 is realized to be used in clinic.

The invention provides an application of a gene IKZF3 in preparing a lung cancer diagnosis kit and the kit. The kit is composed of five parts, including a Real-Time PCR (Polymerase Chain Reaction) primer, a DNA (Desoxvribose Nucleic Acid) standard substance, an RNA (Ribose Nucleic Acid) extraction system, an RNA reverse transcription system and a Real-Time PCR system. The kit is used for analyzing the tumor tissue of a clinical lung cancer patient, detecting the relative expression quantity of the target gene IKZF3 relative to a reference gene GAPDH (reduced glyceraldehyde-phosphate dehydrogenase) in a sample and thus judging the prognosis of the patient.

The invention discloses a stably expressed reference gene group of harmonia axyridis with different factors and application of the reference gene group. Specifically, a scheme of a stably expressed reference gene group of harmonia axyridis with different factors is that a stably expressed reference gene group under different temperature conditions comprises 18S, 28S and GAPDH (Reduced Glyceraldehyde-Phosphate Dehydrogenase), comprises 18S, 28S and HSP90 under different light cycle conditions, comprises 18S, 28S and HSP70 at different age stages, and comprises 18S, 28S and RP49 at different tissue combinations. By adopting the reference gene group, a most stable reference gene group of harmonia axyridis under four factors is confirmed for a first time, reference gene selection bases are made for study on gene expression levels of different tissues of different ages of harmonia axyridis under different temperature conditions and different light cycle conditions, and a solid base is madefor studying gene functions of the harmonia axyridis, RNAi (Ribonucleic Acid Interfere) transgenic crop environment risk estimation, and the like.

The invention discloses a kit and a method for measuring the content of 1,3-diphosphoglycerate based on a micromethod. A reagent of the kit comprises an extracting solution, a first reagent and a second reagent, wherein the extracting solution is prepared from Tris-HCl, EGTA, mercaptoethanol and glycerinum in a mixed mode, the first reagent is prepared from MgSO4.7H2O, NADH, ATP and glyceraldehyde-3-phosphate dehydrogenase in a mixed mode, and the second reagent is prepared from Tris-HCl. The method comprises the principle that the glyceraldehyde-3-phosphate dehydrogenase can reversely catalyze the 1,3-diphosphoglycerate and the NADH to generate glyceraldehyde-3-phosphate, inorganic phosphorus and NAD, the glyceraldehyde-3-phosphate dehydrogenase and the NADH are added into a reaction system, and by measuring the reduction amount of the NADH at 340 nm, the content of the 1,3-diphosphoglycerate can be reflected. The kit and the method for measuring the content of the 1,3-diphosphoglycerate have the characteristics that the application range is wide, operation is easy and convenient, the detection time is short, detection sensitivity is high, the cost is low, repeatability is good, and the accuracy rate is high.