671 results about "Phosphorylation" patented technology

The invention provides a uracil production strain and a construction method and application thereof.According to the strain, on an E.coli UR14 genome by means of a CRIPSR / Cas9 gene editing technology, firstly, psuG genes, preTA genes, rutA genes and upp genes are knocked out, so that decomposition of uracil is blocked; then, a uridine phosphorylase gene udp and a pyrimidine-5 '-nucleotide nucleotidase gene ppnN are subjected to overexpression, and synthesis and accumulation of uracil are synergistically enhanced; and finally, overexpression of the ribose phosphate mutase gene pgm further enhances the conversion of a by-product ribose phosphate 1-precursor 5-ribose phosphate 1-pyrophosphate and improves the carbon utilization rate, and the obtained strain has good genetic stability and high fermentation yield, can stably produce uracil, and has wide application prospects.

The invention discloses a kit for jointly detecting phosphorylated Tau-217 and non-phosphorylated Tau-217. The kit comprises a magnetic microsphere reagent, a detection antibody reagent and a fluorescent microsphere reagent, the magnetic microsphere reagent comprises a magnetic microsphere compound, the detection antibody reagent comprises a first detection antibody compound and a second detection antibody compound, and the fluorescent microsphere reagent comprises a first fluorescent microsphere compound and a second fluorescent microsphere compound. The invention provides a kit for jointly detecting phosphorylated Tau-217 and non-phosphorylated Tau-217, which is based on a single molecule detection technology, and realizes accurate distinguishing and high-sensitivity detection on the phosphorylated Tau-217 and the non-phosphorylated Tau-217 at the same time through a double-target synchronous detection design and a double-color fluorescence report system.

The invention relates to isolated synthetic or recombinant peptides comprising an epitope of human tau 2N4R, wherein the tau peptide sequence comprising the epitope is not phosphorylated. The invention also relates to use of such peptides to generate binding molecules, such as antibodies, specific for the non-phosphorylated tau epitopes and to such peptides and binding molecules, such as antibodies, for use in investigation, diagnosis and treatment of tauopathies, such as Alzheimer's disease.

The invention belongs to the field of biological medicine, and particularly relates to PGK1 K17 site succinylation modification and application. Experiments show that the 17-site lysine of the PGK1 protein has succinylation modification (PGK1 K17su), and the existence of the modification is confirmed through a specific succinylation modified antibody for the first time. Further, it is found that PGK1 K17su can weaken lung cancer cell oxidative phosphorylation and promote glycolysis metabolism by inhibiting lung cancer cell mitochondrial generation, and functional experiments prove that PGK1 K17 succinylation modification promotes proliferation and migration of lung cancer cells.

The invention belongs to the field of antibodies, and particularly relates to a p-Tau217 phosphorylated protein binding antibody or an antigen binding part thereof and application thereof. The invention provides the amino acid sequences of the heavy chain CDR and the light chain CDR of the antibody specifically bound with the p-Tau217 phosphorylated protein, and the antibody can specifically recognize and bind the p-Tau217 phosphorylated protein and has the advantages of high affinity and good specificity to the protein. Based on the advantages, the antibody can be applied to diagnosis of diseases (such as nervous system degenerative diseases) related to abnormal p-Tau217 phosphorylated protein level, and especially can be applied to clinical diagnosis of Alzheimer's disease.

The invention discloses a synthesis method of 2 '-fluoro-2'-deoxyguanosine. According to the method, thymine nucleoside phosphorylase and purine nucleoside phosphorylase are used as biocatalysts, and 2 '-fluoro-2'-deoxyuridine and guanine are used as substrates to synthesize the 2 '-fluoro-2'-deoxyguanosine. Compared with a chemical method, the production cost is greatly reduced; meanwhile, the synthesis method can effectively solve the problem that the product is difficult to separate.

The invention discloses recombinant escherichia coli with high yield of N-acetylneuraminic acid and application of the recombinant escherichia coli, and relates to the technical field of biological genetic engineering. The invention relates to a recombinant escherichia coli, which is characterized in that the escherichia coli is taken as a host, and free expression of an N-acetylmannosamine epimerase gene yihS from Streptomyces xiamenensis or an N-acetylmannosamine epimerase gene ce3 from Bacteroides polymorpha and an exogenous N-acetylneuraminic acid lyase gene nano A is carried out; and carrying out recombinant expression on N-acetyl hexosamine 1-kinase nahK, a UDP-N-acetyl glucosamine pyrophosphorylase gene glmU and a UDP-N-acetyl glucosamine-2-epimerase gene neuC in the other synthetic route of the ManNAc. According to the recombinant escherichia coli with high yield of N-acetylneuraminic acid, the yield of N-acetylneuraminic acid can reach 23.08 g / L under a shake flask fermentation condition; a two-stage batch feeding strategy is adopted, the yield of N-acetylneuraminic acid in a 5L fermentation tank reaches 71.25 g / L, the molar conversion rate of GlcNAc reaches up to 57.60%, and the method has the potential of industrial application.

The invention discloses recombinant bacillus subtilis with high yield of surfactin as well as a construction method and application of the recombinant bacillus subtilis, and belongs to the technical field of genetic engineering. According to the invention, non-essential genes are determined through analysis and identification, a simplified engineering strain is obtained, and the highest fermentation yield reaches 11.2 g / L; then, a PTS part sugar phosphorylase gene obtained through screening is transferred into the simplified engineering strain, the metabolism efficiency of carbohydrates is improved, transportation and utilization of the carbohydrates are optimized, a fatty acid transmembrane transporter gene is further transferred, fatty acid uptake is assisted, synthesis of surfactin is facilitated, and the yield of the surfactin is improved. The combination of the two realizes optimization of synthesis of surfactin through a composite carbon source. Compared with a simplified engineering strain, the genetically engineered bacterium obtained by the invention has the advantages that the yield of surfactin is increased by 57.1%, the yield is increased by 2.3 times compared with that of an unsimplified engineering strain EX, and the final shake-flask fermentation yield can reach 17.6 g / L. As an industrial chassis cell of high-yield lipopeptide, the cell has a good industrial application prospect.

The invention belongs to the technical field of biological materials and biological information, and provides a space phosphorylation modification omics detection method. According to the detection method disclosed by the invention, proteomics analysis can be carried out on trace sample phosphorylation, 4259 phosphorylation sites can be identified by 5 micrograms of peptide fragments, and the credibility of 3506 sites is greater than 0.75.

The invention discloses a chemical-enzymatic coupling synthesis method of oseltamivir phosphate, which comprises the following steps: by taking monasic acid OS-01 as a substrate, sequentially carrying out S1 esterification, S2 ketalation, S3 selective ring opening, S4 TEMPO oxidation, S5 primary transaminase reaction, S6 acetylation, S7 secondary transaminase reaction and S8 phosphorylation reaction to obtain the final product oseltamivir phosphate OS-09. According to the method, the step of sodium azide required by a traditional route is avoided, the production safety and the industrialization possibility are enhanced, two different transaminases are used for respectively constructing different chiral centers, the steps of protecting groups and deprotecting groups on the traditional route are not needed, the overall route is short, the used chemical reagents are few, the yield is high, and the method is suitable for industrial production. The industrial requirements of environmental protection and low cost are met.

The invention discloses a monoclonal antibody which is specifically combined with a microtubule associated protein tau (microtubule associated protein with 217 Thrphorylation, pTau217) of which the 217 site is phosphorylated by threonine, and a preparation method of the monoclonal antibody. The invention provides an amino acid sequence of a variable domain of the monoclonal antibody, and an amino acid sequence of a complementary determining region (CDR) contained in the amino acid sequence of the variable domain of the monoclonal antibody. The monoclonal antibody disclosed by the invention has the capability of specifically recognizing and remarkably combining pTau217, and can be used for developing and producing pTau217 clinical in vitro diagnosis (IVD) medical equipment products based on an immunological detection principle as a reagent raw material.

The invention discloses a formula for reducing phlegm and resolving masses, and a preparation method and application of freeze-dried powder of the formula for reducing phlegm and resolving masses, and belongs to the technical field of traditional Chinese medicines. The traditional Chinese medicine composition inhibits triple negative breast cancer adhesion-metastasis by regulating an FAK / SRC pathway, and comprises the following components by weight: 10-20 g of poria cocos, 10-20 g of rhizoma bolbostemmae, 10-20 g of radix trichosanthis, 5-12 g of curcuma zedoary, 10-20 g of selfheal, 10-20 g of oyster (smashed and decocted first), and 5-12 g of uniflower swisscentaury root. In-vitro experiments show that HTSJF can inhibit TNBC cell invasion, migration and adhesion (P is less than 0.01) in a dose-dependent manner, and along with reduction of the phosphorylation level of an FAK / SRC pathway, it is prompted that HTSJF inhibits TNBC transfer by regulating the pathway. In-vivo experiments prove that the HTSJF can obviously inhibit the reduction of TNBC pulmonary metastasis focus (P is less than 0.05) and recover part of normal pulmonary alveolar structures, and the inhibition effect of the HTSJF on TNBC pulmonary metastasis is further proved.

The embodiment of the specification provides a modification identification method of a thio-phosphorylated nucleic acid sequence, which comprises: performing enzymolysis on a to-be-identified nucleic acid sequence by using a nucleic acid mixed enzyme to obtain an enzymolysis product; performing secondary mass spectrum analysis on the enzymolysis product based on first mass spectrum information; and determining whether a target sequence fragment exists in the to-be-identified nucleic acid sequence based on the first mass spectrum information and the second mass spectrum information. At least the to-be-identified nucleic acid sequence modified by thio-phosphorylation is actually obtained based on a preset modification rule. The first mass spectrum information comprises a theoretical analysis result generated by analyzing a theoretical sequence fragment, and the theoretical sequence fragment is determined based on the preset modification rule, the to-be-identified nucleic acid sequence and the nucleic acid mixed enzyme. The modification identification method can accurately and efficiently identify the chemical modification of the thio-phosphorylated nucleic acid sequence.

The invention discloses a primer probe set for qualitative detection of mycoplasma pneumoniae, an RAA-CHA detection kit and application, and belongs to the technical field of microbiological detection.The primer probe set comprises an upstream primer, a downstream primer, an upstream probe, a downstream probe and a downstream probe, the nucleotide sequence of the upstream primer is as shown in SEQ ID NO: 1, and the nucleotide sequence of the downstream primer is as shown in SEQ ID NO: 2; the nucleotide sequence of the downstream primer is as shown in SEQ ID NO: 2, and the 5'end of the downstream primer is modified by phosphorylation; the nucleotide sequence of the H1 hairpin probe is as shown in SEQ ID NO: 3, and two ends of the H1 hairpin probe are respectively modified with a fluorescence reporter group and a fluorescence quenching group; the nucleotide sequence of the H2 hairpin probe is as shown in SEQ ID NO: 4. According to the application, efficient, accurate and sensitive qualitative detection of the mycoplasma pneumoniae gene is realized through effective amplification and specific recognition of the mycoplasma pneumoniae gene.

Provided herein are novel materials and methods for site-specific incorporation of phosphotyrosines into proteins. The novel methods of the invention encompass the use of a novel aminoacyl tRNA synthetase capable of charging compatible tRNAs with a phosphotyrosine precursor. The phosphotyrosine precursor is then incorporated, site-specifically, into a protein at sites where phosphotyrosine residues are desired. The phosphotyrosine precursors are subsequently treated to convert them into phosphotyrosine residues, yielding proteins with phosphotyrosines at selected sites. The scope of the invention encompasses novel aminoacyl tRNA synthetases, novel phosphotyrosine precursors, and methods of using these materials to create site-specific phosphorylated tyrosine residues in a protein.

A YL1-tiRNA for promoting mouse muscle cell injury repair belongs to the technical field of molecular biology, the YL1-tiRNA is formed by cutting mature mt-Tv, a phosphate group is added at the 5th terminal, phosphorylation modification is carried out on the sequence, and a section of sequence is added at the 5th terminal without changing the function of the sequence, the YL1-tiRNA is characterized in that the nucleotide sequence of the YL1-tiRNA is shown as Seq ID No: 1, and the nucleotide sequence of the YL1-tiRNA is shown as Seq ID No: 1. Experiments prove that the YL1-tiRNA can promote mouse muscle injury repair by regulating proliferation of C2C12 cells.

The invention belongs to the technical field of organic synthesis, and relates to a preparation method of N-palmitoyl-O-phosphorylcholine serine and a deuterated substance thereof. The N-palmitoyl-O-phosphorylcholine serine and the deuterated substance thereof are obtained by taking L-serine methyl ester hydrochloride and palmitic acid as raw materials through amidation reaction, phosphate esterification reaction, quaternization reaction, oxidation reaction, reduction reaction and the like. The invention provides an approach for industrial production of the N-palmitoyl-O-phosphorylcholine serine and the deuterated substance thereof, and is very important for standardization of the detection method of the N-palmitoyl-O-phosphorylcholine serine.

The invention relates to a process for dephosphorylation of skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI) comprising (i) acidification of a skim milk, UF milk or MCI preferably not lower than pH 6.0, preferably between 6.0 and 6.7, (ii) cooling the acidified skim milk, UF milk or MCI to the temperature between 0° C. and 15° C., (iii) adding gluconate and / or maleate to the cooled skim milk, UF milk or MCI and, (iv) washing the skim milk, UF milk or MCI, to remove phosphorus, thus dephosphorylating the skim milk, UF milk or MCI, preferably to an extent that the total phosphorous content of the skim milk, UF milk or MCI is reduced with at least 20%, preferably 20-40%, more preferably 30-40% compared to the material provided to step (i). The invention also relates to dephosphorylated MPC, MPI or MCI obtainable by the process of the invention, and to a liquid heat-sterilized enteral nutritional composition comprising 2.0-3.0 kcal / ml wherein 16-35 en % is provided by protein, the combination of caloric content and relative protein caloric content selected such that there is 10-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein in the composition, wherein the protein comprises micellar casein (MC), whey protein (WP) and optionally caseinate (CAS), wherein there is at least 70 wt % MC and less than 15 wt % WP, based on total protein content, and wherein the composition has a total amount of phosphorous less than 192 mg / 100 ml and / or 30-80 mg / 100 kcal, preferably at least 30-80 mg / 100 kcal, most preferably less than 192 mg / 100 ml and 30-80 mg / 100 kcal.

The present invention provides a therapeutic agent for sepsis and / or septic shock, comprising, as an active ingredient, a compound capable of suppressing phosphorylation of threonine at position 749 in human STAT1; a method for screening for a candidate compound serving as an active ingredient of a therapeutic agent for sepsis and / or septic shock, the method comprising selecting a compound capable of suppressing phosphorylation of threonine at position 749 in human STAT1; a therapeutic agent for colitis, comprising, as an active ingredient, a compound capable of promoting phosphorylation of threonine at position 749 in human STAT1; a method for screening for a candidate compound serving as an active ingredient of a therapeutic agent for colitis, the method comprising selecting a compound capable of promoting phosphorylation of threonine at position 749 in human STAT1; a therapeutic agent for systemic lupus erythematosus, comprising, as an active ingredient, a compound capable of inhibiting human STAT1; and a method for screening for a candidate compound serving as an active ingredient of a therapeutic agent for systemic lupus erythematosus, the method comprising selecting a compound capable of inhibiting human STAT1.

The invention relates to the technical field of biological medicines, and particularly discloses synthesis and application of oligonucleotide with a spacer group embedded between 5-terminal oxygen and thiophosphoric acid. The chemical modification strategy comprises the following steps: preparing a modified nucleotide phosphoramidite monomer in which a spacer group is embedded between O5'and thiophosphoric acid P (V); performing solid-phase synthesis on the modified nucleotide phosphoramidite monomer to construct a target oligonucleotide molecule; a spacer group is embedded between oxygen at the 5 '-terminal of the oligonucleotide and thiophosphoric acid P (V) to form a 5'-terminal thiophosphoric acid structure; as the terminal modified thiophosphoric acid belongs to a non-phosphatase substrate, the terminal modified thiophosphoric acid can resist exonuclease degradation and improve the biological activity of siRNA after being modified.

The invention relates to the technical field of molecular biology and the technical field of gene editing, in particular to pegRNA of a specific target EGFR gene tyrosine 1068 site, a pilot editing system and application. The pegRNA comprises sgRNA (small guide RNA), sgRNA scaffold, an RT (reverse transcription) template and PBS (phosphate buffer solution); wherein the sequence of the pegRNA is as shown in SEQ ID NO. 1; the sequence of the sgRNA is as shown in SEQ ID NO. 2; the sequence of the sgRNA scaffold is as shown in SEQ ID NO. 3; the sequence of the RT template is as shown in SEQ ID NO. 4; the sequence of the PBS is as shown in SEQ ID NO. 5. The tyrosine 1068 site of the EGFR gene is mutated into phenylalanine, so that phosphorylation of the site is blocked, and a protein structure is maintained. The tyrosine 1068 site point mutant with the EGFR gene can be applied to research on preparation of drugs for treating cancers, research on drug resistance and research on screening of compounds of targeted EGFR signal channels.

The invention relates to the technical field of medicines, in particular to a composition with ingenane diterpenoid compounds as chemotherapeutic drug sensitizers and application of the composition. The chemotherapeutic drug sensitizer and the composition thereof provided by the invention can be used as the chemotherapeutic drug sensitizer, especially the sensitizer of breast cancer treatment drugs, and can be used for increasing the sensitivity of breast cancer cells to the chemotherapeutic drugs by inhibiting CHK1 (Ser345) phosphorylation so as to realize sensitization and provide a new treatment strategy for treating breast cancer.

The application provides a combination drug of a PLA2G7 inhibitor and a chemotherapeutic drug and its use in resisting triple-negative breast cancer, and belongs to the technical field of medicines. The combination drug is a PLA2G7 inhibitor and a chemotherapeutic drug in the same or different specifications of unit preparations for simultaneous or separate administration, and a pharmaceutically acceptable carrier. The application first discovers and proves that the PLA2G7 inhibitor Darapladib can significantly enhance the anti-tumor activity of paclitaxel, 5-fluorouracil or cisplatin on triple-negative breast cancer, and meanwhile, the combination of the PLA2G7 inhibitor and paclitaxel has a synergistic effect in down-regulating the phosphorylation level of STAT3 protein in cancer cells and inhibiting the growth and proliferation of cancer cells; the combination of the PLA2G7 inhibitor and 5-fluorouracil also shows a synergistic effect in inhibiting the growth and proliferation of cancer cells. The application provides a new and effective combination drug strategy for improving the treatment effect of the chemotherapeutic drug on triple-negative breast cancer.

The present invention refers to a method for manufacturing a phosphorylated lignin which is soluble in water at a pH range of 2 to 14. Said method comprises the efficient phosphorylation of lignin preventing the formation of undesired species, which leads to an outstanding water solubility at neutral and acidic conditions, and, preferably, the separation of the water-soluble phosphorylated lignin from the reaction medium, wherein said separation comprises acid precipitation and / or ultrafiltration and drying under specific conditions. The invention also refers to a phosphorylated lignin that is water- soluble at a pH of 2 to 14, preferably obtained by the method of the invention.

The invention discloses an application of a pyruvate dehydrogenase kinase (PDKS) inhibitor in preparation of a composition for treating Alzheimer's disease (AD), and particularly discloses an application of the PDKs inhibitor in preparation of the composition for treating the AD. According to the invention, the level of oxidative phosphorylation (OXPHOS) of microglial cells is obviously enhanced, the inflammatory response of the microglial cells is effectively reduced, the migration and aggregation capabilities of the microglial cells are obviously improved, the capability of the microglial cells to phagocytize and degrade extracellular beta-amyloid protein (A beta) plaque is obviously enhanced, the cognitive function defect of AD model mice is effectively improved, and the application of the microglial cells in the treatment of the AD model mice is promoted. The preparation is simple, and the application is wide.

A device for activating the mitochondrion, wherein negative charge and reactive oxygen are delivered from outside to the mitochondrion via a negative charge and reactive oxygen delivery unit to serve as material for the mitochondrial redox reaction; and, wherein a calcium ion channel stimulation unit can stimulate an in vivo calcium ion to transport a protein across a calcium ion on an inner membrane of the mitochondrion into a matrix space of the mitochondrion, whereby an influx of the calcium ion promotes the depolarization of the mitochondrion and increases the activity of mitochondrial oxidative phosphorylation.

The application relates to the field of myocardial ischemia-reperfusion injury drugs, and discloses application of Gualou Xiebai Baijiu decoction in preparation of myocardial ischemia-reperfusion injury drugs. The myocardial ischemia-reperfusion injury drugs inhibit the GSK-3beta / Drp1-mediated mitochondrial fission pathway by targeting Akt protein, maintain the dynamic balance of mitochondrial dynamics, and play a myocardial protection role. The drugs can promote Akt phosphorylation activation, inhibit GSK-3beta activity, down-regulate Drp1 protein Ser616 site phosphorylation, up-regulate Ser637 site phosphorylation, reduce Drp1 translocation and aggregation to the outer membrane of mitochondria, and simultaneously up-regulate the expression of mitochondrial fusion proteins Mfn1 and Mfn2. The drugs can maintain the structural integrity of mitochondria, improve the mitochondrial membrane potential and ATP content, improve myocardial cell energy metabolism, regulate the expression of apoptosis-related proteins to inhibit myocardial cell apoptosis, improve the cardiac systolic function after ischemia-reperfusion, reduce the myocardial infarction area, reduce the serum myocardial injury marker level, and regulate the oxidative stress state.

This application relates to the field of biomedical technology and discloses the application of juniperol or its derivatives in the preparation of drugs that inhibit Drp1-mediated excessive mitochondrial division. This application verifies through in vitro and in vivo experiments that juniperol can inhibit phosphorylation at the Ser616 site of Drp1 and Drp1 mitochondrial translocation, thereby inhibiting Drp1-mediated excessive mitochondrial division. This demonstrates a good therapeutic effect on mitochondrial excessive division-related diseases such as inflammatory bowel disease, providing a new therapeutic strategy for the treatment of these diseases and possessing significant clinical implications.

The invention discloses application of jellyfish and snow rabbits in inhibition of ERK phosphorylation level and MAPK pathway and an evaluation method of the jellyfish and snow rabbits, and relates to the technical field of biological medicines.The method comprises the following steps that 1, a treated jellyfish and snow rabbits breast cancer cell group and a control breast cancer cell group are established; and step 2, respectively dividing the two groups of samples in the step 1 into two parts, fixing one part in paraformaldehyde with the mass concentration of 10%, treating, embedding in paraffin for immunohistochemical analysis, and quickly freezing the other part for transcriptomics analysis and the like. According to the invention, UPLC-MS is adopted to determine the main components of SMM, the influence of SMM on breast cancer (BC) is evaluated through in-vivo experiments, in order to deeply clarify the mechanism, network pharmacology and transcriptomics analysis are integrated, finally, immunohistochemistry (IHC) is adopted to verify and explore the exact breast cancer resisting mechanism of SMM, and a scientific basis is provided for potential clinical application of SMM.

The present application relates to a kind of 2'-fucosyllactose producing recombinant escherichia coli and its construction method and application, belong to the field of bioengineering technology.The present application expresses 2'-fucosyllactose remediation synthesis pathway related gene in recombinant escherichia coli, including the gene fkp of coding GDP-fucose pyrophosphorylase FKP and the gene futC of coding fucosyltransferase FutC, respectively in the upstream of fkp gene and futC gene expression macromolecular coagulation material protein FUSLCD after the modification of coagulation tag GCN4, and replaces replicon, constructs a strain of high-yield 2'-fucosyllactose recombinant escherichia coli, yield and production intensity reach 38.68g / L and 0.35g / L / h respectively.It has good application prospect, has the potential of industrial application.