73 results about "Glyceraldehyde 3-phosphate" patented technology

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 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.

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.

A pyrophosphoric acid sensor that in the method of measuring pyrophosphoric acid in SNP typing making use of primer extension reaction, realizes convenient detection of pyrophosphoric acid with high sensitivity. There is provided a pyrophosphoric acid sensor composed of insulating substrate (1); formed thereon, an electrode group consisting of measuring electrode (2) and counter electrode (3); and superimposed on the substrate (1), multiple reaction reagent layers consisting of pyrophosphatase, glyceraldehyde-3-phosphate dehydrogenase, diaphorase, glyceraldehyde-3-phosphate, oxidized nicotinamide adenine dinucleotide, electron mediator, magnesium salt and buffer solution components wherein reaction reagent layer (35) containing buffer solution components is separated from enzyme-containing reaction reagent layer (36), characterized in that reaction reagent layer (37) containing glyceraldehyde-3-phosphate is separated from the reaction reagent layer (35) containing buffer solution components.

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.