73 results about "Glyceraldehyde 3-phosphate" patented technology

The invention provides a method for synthesizing isoprene from pyruvic acid and 3-phosphoglyceraldehyde, and a reconstituted cell. The pyruvic acid and the 3-phosphoglyceraldehyde are finally obtained from simple starting materials such as glucose.

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 provides a Ti plasmid aspergillus niger gene replacement expression vector and application thereof, and belongs to the technical field of molecular biology. The T-DNA (Triple helix Deoxyribose Nucleic Acid) region elements of the Ti plasmid aspergillus niger gene replacement expression vector are arranged in the following sequence: an aspergillus niger target gene promoter, a multiple cloning site, an aspergillus niger target gene terminator, an aspergillus nidulans 3-phosphoglyceraldehyde dehydrogenase gene promoter PgpdA, an aspergillus niger selection marker gene and an aspergillus niger target gene terminator. According to the invention, a target gene is integrated at the site of the aspergillus niger target gene through homologous recombination, and the target gene is regulated and controlled by a target gene promotor of high expression; therefore, the position effect of transgenosis is eliminated and the expression level is improved.

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 present invention relates to an integrated protein C-terminal enrichment method, which comprises: selective dimethylation labeling, DL-glyceraldehyde-3-phosphate labeling and affinity removing with titania. According to the method, an enzymolysis product of a protein sample is subjected to selective N-terminal amino dimethylation labeling on a reverse phase trapping column under an acid condition, the side chain amino of the peptide segment is labeled on a DL-glyceraldehyde-3-phosphate column, and finally a TiO2 material is adopted to carry out affinity removing to remove the phosphate-labeled peptide segment so as to obtain the protein C-terminal, wherein sample transfer and desalination lyophilization are not required to be performed during the process. The method has advantages of high labeling efficiency, high selectivity, simple treatment steps, time saving, sample consumption saving, and sample loss and contamination reducing. In addition, the method provides good compatibility with the chromatography-mass spectrometry separation and identification platform.

The application of the invention provides a construction method of an efficiently-expressed plasmid for producing lipase gene. The method comprises the following steps: cloning an obtained hygromycin-resistant expression box on an objective plasmid so as to obtain a hygromycin-resistant recombinant plasmid; the respectively amplifying lipase gene (PEL) of blue mould, aspergillus nidulans strong promoter 3-phosphoglyceraldehydedehydrogenase promoter (PgpdA) and aspergillus nidulans tryptophan synzyme terminator (TtrpC) by PCR (polymerase chain reaction) technology to obtain a PEL gene expression box driven by a strong promoter; and inserting the PEL gene expression box into the hygromycin-resistant recombinant plasmid to obtain a hygromycin selection marker-containing PEL gene overexpression vector. The efficiently-expressed plasmid for producing lipase gene is transferred to obtain the genetic engineering penicillium of the efficiently-expressed plasmid, high-efficiency and stable in expression. Compared with conventional lipase production method, the construction method has great advantages.

Aiming at the drawbacks of the prior art, the invention applies for providing a method for breeding a penicillium gene engineering bacterium having high lipase synthesis capability. The method comprises: obtaining a hygromycin-resistance expression box and cloning to a target plasmid to obtain a hygromycin-resistance plasmid; amplifying the lipase gene, namely primary effusion lymphoma (PEL), of the penicillium and the strong promoter, namely glyceraldehyde-3-phosphate dehydrogenase promoter PgpdA, of Aspergillus nidulans and tryptophan synthetase terminator TtrpC of the Aspergillus nidulans respectively to obtain a PEL gene expression box driven by the strong promoter; inserting the PEL gene expression box into the hygromycin-resistance plasmid to obtain a PEL gene super expression vector containing a hygromycin screening marker; and finally, transferring Agrobacterium rhizogenes by using the PEL gene super expression vector containing the hygromycin screening marker, and transferring the PEL gene expression box into a penicillium strain by using a Agrobacterium rhizogenes-mediated transfer method to obtain the penicillium gene engineering bacterium with high lipase expressing capability.

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 preparation method of 4-phosphoric acid erythrose, comprising the following steps: immobilizing heat-resistant transketolase at the temperature of 55-65 DEG C; and then taking 6-fructose phosphate and 3-glyceraldehyde phosphate as raw materials to prepare the 4-phosphoric acid erythrose by enzymatic reaction. In the preparation method, the heat-resistant and specifically transformed transketolase of the 4-phosphoric acid erythrose is prepared by adopting a molecular biology technology, and the 6-fructose phosphate and the 3-glyceraldehyde phosphate are taken as the raw materials to efficiently prepare the 4-phosphoric acid erythrose by adopting an immobilized enzyme technology. The preparation method is simple and practical.

The invention relates to a method for synthesizing acrylic acid from cyanobacteria. The method comprises the following steps: 1), firstly, based on an NSI gene of cyanobacteria Syn7942, constructing an integrated vector NSI, and recombining to obtain a vector NSI-ceaS2; 2), transforming the recombinant vector NSI-ceaS2 into cyanobacteria Syn7942 cells, and then preliminarily screening out monoclonal transgenic cyanobacteria Syn7942 through a chloramphenicol solid culture medium; 3), transferring the screened monoclonal transgenic cyanobacteria Syn7942 to a liquid culture medium with chloramphenicol resistance, and after the cyanobacteria grow out, extracting a cyanobacteria genome for PCR verification of a target gene; 4), transferring the successfully verified monoclonal transgenic cyanobacteria Syn7942 to the liquid culture medium for culture, and when the cyanobacteria grow till OD730 is greater than or equal to 1, adding IPTG for inducing expression of a gene ceaS2, and catalytically synthesizing the acrylic acid by an enzyme by using glyceraldehyde-3-phosphate (G3P) or dihydroxyacetone phosphate (DHAP) produced by photosynthesis of the cyanobacteria as a substrate; 5), performing separation and purification. By the method, when the acrylic acid is synthesized through transformation of the cyanobacteria as substrate organisms, only sunlight and moisture and the like are required as production raw materials, production equipment and a production environment are easy to construct, and high electricity consumption is not required, so that the production cost of the methodprovided by the invention is greatly lower than those of other production methods.

The invention discloses p pichia yeast bacterium producing aspergillus-niger glucose oxidase, a construction method and application thereof, and belongs to the technical field of gene engineering. A recombined pichia yeast GS1168-GOD capable of producing glucose oxidase with relatively high activity is obtained by using a recombination DNA technology to clone and link glucose oxidase (GOD) gene of aspergillus niger accc30161 is cloned to pichia-yeast secretion-type expression vector pGAPZalphaA containing glyceraldehydes-3-phosphate dehydrogenase gene promoter, transforming pichia pastoris GS1168 and performing screening and identification. The enzyme activity of glucose oxidase expressed by the bacterial strain under the regulation and control of glyceraldehydes-3-phosphate dehydrogenase gene promoter reaches 107 U/mL, and a good base is established for large-scale production and application of glucose oxidase.

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.

The invention relates to a kit for diagnosing/mensurating carbon dioxide by utilizing the technologies of the enzymic colorimetric method and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for mensurating the concentration of the carbon dioxide, and belongs to the technology field of medical/food/environmental inspection and measurement. The main components of the kit include a buffer solution, coenzyme, phosphoenolpyruvic acid, glyceraldehyde-3-phosphate, glyceraldehyde-3-phosphate dehydrogenase, phosphoenol pyruvate carboxylase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, an enzymatic reactions occurs, then the reactant is placed under an ultraviolet/visible light analyzer, and the degree/velocity of the increase in absorbance at 340nm of the dominant wavelength is detected, thereby mensurating the concentration of the carbon dioxide.

This invention provides an anti-allergic substance that effectively inhibits allergic symptoms by inhibiting production of the IgE antibody associated with the development of allergies. Anti-allergic substances, pharmaceutical products (i.e., anti-allergic agents), and food (i.e., food with health-promoting benefits) containing strawberry-derived glyceraldehyde-3-phosphate do not cause side effects such as those caused by steroids, and such substances can be consumed through daily meals and can alleviate allergic symptoms.

The invention discloses p pichia yeast bacterium producing aspergillus-niger glucose oxidase, a construction method and application thereof, and belongs to the technical field of gene engineering. A recombined pichia yeast KM71-GOD capable of producing glucose oxidase with relatively high activity is obtained by using a recombination DNA technology to clone and link glucose oxidase (GOD) gene of aspergillus niger accc30161 is cloned to pichia-yeast secretion-type expression vector pGAPZalphaA containing glyceraldehydes-3-phosphate dehydrogenase gene promoter, transforming pichia pastoris KM71 and performing screening and identification. The enzyme activity of glucose oxidase expressed by the bacterial strain under the regulation and control of glyceraldehydes-3-phosphate dehydrogenase gene promoter reaches 107 U/mL, and a good base is established for large-scale production and application of glucose oxidase.

The invention relates to a homocysteine diagnosing/measuring reagent (kit) using the technologies of an enzyme colorimetric method and an enzyme linkage method. At the same time, the invention also relates to a homocysteine concentration measuring method, the reagent composition and reagent ingredients, which belong to the technical field of medical detection and measurement. The reagent (kit) mainly comprises the following ingredients: a buffer solution, cozymase, 5-methyl tetrahydrofolate, formic acid, adenyl pyrophosphate, glyceraldehyde-3-phosphate, methionine synthesis enzyme, formic acid-tetrahydrofolic acid ligase, glyceraldehyde-3-phosphate dehydrogenase and stabilizing agents. The method has the following steps: mixing samples and the reagent by the certain volume percent for making the samples and the reagent take a series of enzymatic reaction, and then, placing reactants under an ultraviolet/visible light analyzer to detect the rising degree of the light absorbancy in the position with the main wave length of 340 nm, so the concentration of the homocysteine can be measured and calculated.

The invention relates to a reagent (kit) for diagnosing/determining amino acid by using an enzyme multiplication method, an enzyme colorimetric method and an enzyme link method, and also discloses a method for determining the concentration of the amino acid, a composition and components of the reagent, belonging to the technical field of medicine/food/environmental test determination. The reagent (kit) comprises the main components: a buffer solution, a coenzyme, sodium bicarbonate (carbon dioxide), adenosine triphosphoric acid, glyceraldehyde-3-phosphate, an amino acid dehydrogenase, a carbamyl phosphate synthetase, a glyceraldehyde-3-phosphate dehydrogenase and a stabilizing agent. A sample and the reagent are mixed according to a certain volume ratio to generate a series of enzymatic reactions, and reactants are placed under an ultraviolet/visible light analyzer for detecting the degree of absorbance rise at the dominant wavelength of 340 nm, thereby measuring and calculating the concentration of the amino acid.