121 results about "Glyceraldehyde-3-Phosphate Dehydrogenases" patented technology

The present invention relates to a method for analyzing and rapidly determining the quality of a test RNA of unknown quality utilizing a novel quantitative reverse transcription-polymerase chain reaction (RT-PCR) based method. The present invention is based on normalizing the 3′ end of the house-keeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to the relative abundance of the 5′ end of GAPDH MRNA in the test RNA. The present invention is particularly useful in pre-screening postmortem tissue samples for microarray experiments and for evaluating large quantities of samples which would be time consuming and expensive to analyze by methods currently in use.

The regulatory sequences associated with the Yarrowia lipolytica glyceraldehyde-3-phosphate dehydrogenase (gpd) and phosphoglycerate mutase (gpm) genes have been found to be particularly effective for the expression of heterologous genes in oleaginous yeast. The promoter regions of the invention, intron and enhancer have been shown to drive high-level expression of genes involved in the production of ω-3 and ω-6 fatty acids.

Genetically engineered Streptomyces clavuligerus strains with improved capabilities to produce clavulanic acid are provided. The strains are genetically engineered by disrupting newly identified glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes. This results in an increased intracelluar pool of the clavulanic acid precursor D-glyceraldehyde-3-phosphate (D-G3P), and increased clavulanic acid production. Clavulanic acid production may be further increased by supplying arginine to the medium in which the S. clavuligerus is grown.

The invention discloses a preparation method of a Trichoderma reesei gene engineering bacterium for highly yielding heat stability xylanase. The method comprises the following steps: synthesizing an expression cassette sequence sequentially comprising a Trichoderma reesei glyceraldehyde-3-phosphate dehydrogenase promoter, a Trichoderma reesei cbh1 signal peptide sequence, an xylanase xyn-LVK gene sequence and a Trichoderma reesei glyceraldehyde-3-phosphate dehydrogenase terminator; cloning the expression cassette sequence into pUC19 in order to obtain a recombinant expression plasmid; and lightly uniformly mixing the recombinant expression plasmid with a pAN7-1 vector, co-transforming Trichoderma reesei protoplasts, and screening the Trichoderma reesei protoplasts to obtain the Trichoderma reesei gene engineering bacterium for highly yielding heat stability xylanase xyn-LVK. The content of the xylanase xyn-LVK expressed in the Trichoderma reesei is about 40% higher than the content of the xylanase xyn-LVK expressed in Pichia yeast.

The invention relates to the field of genetic engineering, particularly to a method for realizing high expression of CP4 (cysteine proteinase 4)-EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) fusion protein gene in a constitutive manner in Hansenula polymorpha cells by using a composite enhancer. The method comprises the following steps: (1) improving the yield of codon-optimized CP4-EPSPS fusion protein gene in Hansenula polymorpha by using the composite enhancer; (2) regulating and controlling CP4-EPSPS fusion protein gene to express in a constitutive manner in polytype Hansenula by adopting Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase promoter; (3) fermenting optimized Hansenula polymorpha; and (4) rapidly purifying the recombinant heterologous protein by employing nanofiltration and nickel column affinity chromatography. The CP4-EPSPS recombinant fusion protein prepared by the method can serve as an alternative protein standard substance in the field of study on the origin of protein and the like.

According to the present invention, a series of genes are identified in Group B Streptococcus, the products of which may be located on the outer surface of the organism. The genes, or functional fragments thereof, may be useful in the preparation of therapeutics, e.g. vaccines for the immunization of a patient against microbial infection.

A sensor of pyrophosphate which can detect pyrophosphate conveniently with high sensitivity in a method for measuring pyrophosphate in SNP typing utilizing a primer extension reaction is provided.A sensor of pyrophosphate which is characterized by including: an insulative substrate 1; an electrode system that is formed thereon and has a measurement electrode 2 and a counter electrode 3; and a plurality of reaction reagent layers that are provided on the substrate 1 and include pyrophosphatase, glyceraldehyde-3-phosphate dehydrogenase, diaphorase, glyceraldehyde-3-phosphate, oxidized nicotineamide adenine dinucleotide, an electronic mediator, a magnesium salt and a buffer component, reaction reagent layer 36 including the enzyme being separated from reaction reagent layer 35 including the buffer component, and reaction reagent layer 37 including glyceraldehyde-3-phosphate being separated from the reaction reagent layer 35 including the buffer component.

The invention relates to a triple fluorescence PCR method for detecting bovine-derived, sheep-derived and porcine-derived ingredients. The method comprises four elements, namely a sample DNA, a pair of universal primers and three probes, fluorescence PCR premixed liquid as well as a positive control. The method comprises the following steps: designing primers and probes according to glyceraldehyde-3-phosphate dehydrogenase genes of cows, sheep and goats as well as pigs, amplifying a target sequence, and exciting and quenching a fluorescence signal, wherein a non-target sequence is not amplified and has no fluorescence signal; and preparing a kit according to a formula of a reagent used in fluorescence PCR amplification carried out on a to-be-detected sample DNA and amplification conditions. Concrete actions are as follows: a triple fluorescence PCR primer system used for detecting the bovine-derived, sheep-derived and porcine-derived ingredients is established; a triple fluorescence PCR kit used for detecting the bovine-derived, sheep-derived and porcine-derived ingredients is established; and a triple fluorescence PCR identification method used for detecting the bovine-derived, sheep-derived and porcine-derived ingredients is determined. The method provided by the invention has the advantages that the standard error is small, the detection time is short, multiple target genes can be detected at the same time, and the reagent cost is saved; and the method is applicable to true and false identification of an animal product.

The invention found that partial deletion of the glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter can enhance gene expression (even heterologous gene expression) in basidiomycetous fungi. With the discovery of these gpd promoters, an expression system can be constructed for the expression of a heterologous gene in mushroom. Accordingly, the invention provides a truncated glyceraldehyde-3-phosphate dehydrogenase promoter and a construct comprising the promoter of the invention operably linked to a heterologous transcribable polynucleotide molecule and a mushroom comprising the construct.

The invention relates to discovery of an isolated glyceraldehyde-3-phosphate dehydrogenase promoter in Pleurotus and a construct comprising the promoter of the invention operably linked to a heterologous transcribable polynucleotide molecule.

A sensor of pyrophosphate which can detect pyrophosphate conveniently with high sensitivity in a method for measuring pyrophosphate in SNP typing utilizing a primer extension reaction is provided.A sensor of pyrophosphate which is characterized by including: an insulative substrate 1; an electrode system that is formed thereon and has a measurement electrode 2 and a counter electrode 3; and a plurality of reaction reagent layers that are provided on the substrate 1 and include pyrophosphatase, glyceraldehyde-3-phosphate dehydrogenase, diaphorase, glyceraldehyde-3-phosphate, oxidized nicotineamide adenine dinucleotide, an electronic mediator, a magnesium salt and a buffer component, reaction reagent layer 36 including the enzyme being separated from reaction reagent layer 35 including the buffer component, and reaction reagent layer 37 including glyceraldehyde-3-phosphate being separated from the reaction reagent layer 35 including the buffer component.