140 results about "Acetyltransferase" patented technology

A method for simply and conveniently detecting acetyltransferase and deacetylase activities of proteins by executing an acetylation reaction of a peptide substrate with an acetyltransferase, or a deacetylation reaction of an acetylated peptide substrate with a deacetylase, and after the completion of these reactions, detecting the acetyl group bound to the peptide substrate by using an anti-acetylated peptide antibody. This system for detecting acetyltransferase and deacetylase activities using the anti-acetylated peptide antibody enables screening inhibitors or enhancers of acetyltransferase and deacetylase. A system for screening deacetylase inhibitors or acetyltransferase enhancers using cultured cells is also provided.

Provided are recombinant DNA molecules that do not occur in nature encoding an O-acetyltransferase, vectors that direct expression of an O-acetyltransferase, recombinant host cells which express an O-acetyltransferase, methods for recombinant production of an O-acetyltransferase, methods for acetylating capsular polysaccharides, especially those of a Serogroup A Neisseria meningitidis using a recombinant O-acetyltransferase, and immunogenic compositions comprising the acetylated capsular polysaccharide.

The present invention provides a method for measuring diacylglycerol acetyltransferase (DGAT) activity which utilizes a novel solvent system to reduce and/or eliminate the activities of related compounds. The present invention also discloses a method for determining whether a compound is useful for modulating DGAT biological activity. The method is capable of being utilized for mass screening of compounds as modulators of the biological activity of DGAT.

The invention relates to a detection kit for ureaplasma urealyticum (UU) nucleic acid by utilizing RNA constant-temperature amplification. The detection kit comprises a urine sample preserving solution, a nucleic acid extraction solution, a UU reaction solution, a UU detection solution, an SAT (Spermidine/Spermine N1-Acetyltransferase) enzyme solution, a UU positive control and a UU negative control. The detection kit provided by the invention has the analytical sensitivity of 50 CFU/reaction for detecting UU, and has the characteristics of high specificity and high sensitivity; and an amplification product RNA is easy to degrade in a natural environment and causes low pollution.

The invention discloses an acetyl coenzyme A acetyltransferase gene RKAcaT2 and application thereof. The nucleotide sequence of the gene is shown in SEQ ID NO:1, the amino acid sequence encoded by thegene is shown in SEQ ID NO:2, the gene is a key enzyme gene synthesized by carotenoid in Rhodosporidium kratochvilovae YM25235 and has the functions of the acetyl coenzyme A acetyltransferase, and the carotenoid produced by the Rhodosporidium kratochvilovae YM25235 can be controlled; microorganisms are transformed through the genetic engineering means so as to increase the yield of carotenoid inthe microorganisms, and a foundation is laid for large-scale commercial production of the carotenoid.

The invention discloses an electrochemical method for detecting the activity of protein acetyltransferase. P2 templated silver nanoparticles and cucurbit [8] urea are added into a P1 modified gold electrode and a detection system of a p300 protein, the activity of the p300 protein is detected by electrochemical measurement, and the protein transferase activity is obtained; the amino acid sequenceof P1 is 11-mercaptoundecanoic acid (MUA)-GGGFRGKGGKGLGKGGAKA; and the amino acid sequence of P2 is FGGGASLWWSEKL. p300 is used as a model, and the silver nanoparticles are synthesized by a signal template P2, and assembled with a sensing template P1 through a cavity, so that the sensitivity of detection is ensured, high design flexibility is achieved, and meanwhile, the selectivity of HATs is enhanced. The development of the simple and practical electrochemical method capable of detecting HATs activity is expected to help early diagnosis of major diseases.

The invention discloses a construction method and an application of an electrochemical Faraday cage immunosensor for detecting the activity of histone acetyltransferase. The construction method comprises the following steps: (1) preparing peptide/Au: respectively taking acetyltransferase p300, polypeptide and acetyl-CoA, fully mixing the taken substances in PBS (0.1 M, pH 7.0), incubating the obtained solution in a constant-temperature water bath, taking and dropwise applying a catalytic reaction to the surface of a gold electrode, and incubating the gold electrode in a 4 DEG C refrigerator; (2) preparing MB&AuNPs@GO-Ab: mixing HAuCl4 and CTAB with water, adding ascorbic acid to the obtained reaction mixture, adding NaOH to obtain CTAB covered AuNPs, centrifuging and purifying the CTAB covered AuNPs, dispersing the purified CTAB covered AuNPs in an equal amount of water, adding GO to the obtained solution, performing ultrasonic dispersion, standing the dispersed solution for later use,adding an acetyl antibody, performing incubation, adding MB, and performing vibration and uniform mixing; and (3) producing the electrochemical Faraday cage immunosensor: taking and dropwise applyingthe MB&AuNPs@GO-Ab to the surface of the peptide/Au, performing incubation at room temperature, and placing the produced sensor in the PBS (0.1 M, pH 7.0) to carry out electrochemical SWV test.

The invention discloses a method for constructing a Dunaliella salina chloroplast transformation vector, in particular a method for constructing a dual-exchange dual-selection marker Dunaliella salina chloroplast transformation vector by using a light dependence free protochlorophyllide reductase chlN, chlB or chlL gene as a homological segment, chloramphnicol acetyltransferase (CAT) gene and a resistance gene bar of phosphinothricin (PPT) as a weedicide as screening markers and an atpA promoter and a rbcL terminator as expression elements. The Dunaliella salina is transformed through a gene gun method or electrization method and then subjected to two-step screening to realize homogenization, fixed point integration of an exogenous gene in a Dunaliella salina chloroplast genome is ensured, and finally the chloroplast transformation Dunaliella salina strain for stably expressing the exogenous gene is obtained. In the invention, the chlN, chlB or chlL gene is selected as the homological segment and can be used as an auxiliary screening marker of a transformant; and the chlorampenicol resistant CAT gene and the resistance gene bar of the PPT are selected to be used as screening marker genes together, thus the stable transformation strain can be obtained within shorter time.

The invention discloses a construction method of an engineering strain for producing tetrahydropyrimidine by a biological method and application of the engineering strain in tetrahydropyrimidine production. According to the engineering strain provided by the invention, heterologous expression of an tetrahydropyrimidine synthetic gene cluster of a marine microorganism salinicola salinia is utilized, so that aminobutyric acid acetyltransferase, diaminobutyric acid aminotransferase and tetrahydropyrimidine synthetase are highly expressed in series in escherichia coli, and then sodium aspartate is catalyzed through an enzymatic reaction to generate tetrahydropyrimidine. The bioconversion method of the tetrahydropyrimidine has the characteristics of mild reaction temperature, simple process and high conversion rate.

The invention discloses multipath composite neuraminic acid producing bacillus subtilis and an application thereof, and belongs to the field of genetic engineering. The expression levels of key enzymes, namely UDP-N-acetylglucosamine-2-epimerase, N-acetylglucosamine-6-phosphate isomerase, glucosamine-6-phosphoric acid-N-acetyltransferase, N-acetylglucosamine isomerase and N-acetylneuraminate synthase on genomes in three neuraminic acid synthesis paths are sequentially optimized by 10 promoters with different intensities, so that the protein synthesis pressure of enzyme expression on cells is reduced; and three neuraminic acids are further integrated into the same engineering bacillus subtilis, so that the bacillus subtilis with the increased N-acetylneuraminic acid yield is obtained, the neuraminic acid yield under the shake flask level reaches 10.4 g/L, and a foundation is laid for further increasing the NeuAc yield of the bacillus subtilis.

The invention discloses an enhance-like element gene ER1 for enhancing foreign protein expression in prokaryotic cells. The enhance-like element gene has a nucleotide sequence as shown in SEQ ID NO: 1 (Sequence Identity No) or a truncated sequence of the nucleotide sequence, wherein the gene sequence is screened by establishing a fusion expression reporter gene recombinant vector; the fusion reporter gene consists of major capsid protein L1 gene of human papilloma virus and chloramphnicol acetyltransferase (CAT); the gene fragments to be selected are linked with the sieving recombinant vector and is transformed so as to establish a gene library; and the recombinant bacterial strain containing the enhance-like element gene sequence is screened based on the chloramphenicol resistance, so as to obtain the enhance-like element sequence which can improve the activity of chloramphenicol of the fusion reporter gene of the strain and promotes the foreign protein expression.

The invention discloses nearly 480 novel phosphorylation sites identified in signal transduction proteins and pathways underlying human Leukemia, and provides phosphorylation site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: adaptor/scaffold proteins, acetyltransferases, actin binding proteins, adhesion proteins, apoptosis proteins, calcium-binding proteins, cell cycle regulation proteins, cell surface proteins, channel proteins, chaperone proteins, contractile proteins, cytokine proteins, cytoskeletal proteins, G protein regulators and GTPase activating proteins, guanine nucleotide exchange factors, helicase proteins, immunoglobulin superfamily proteins, inhibitor proteins, protein kinases, lipid kinases, ligases, lipid binding proteins, methytransferases, motor proteins, oxidoreductases, phosphotases, phosphodiesterases, phospholipases, proteases, receptor proteins, trascription factors, transferases, translation/transporter proteins, and ubiquitin conjugating system proteins.

A bivalent plant expression carrier for culturing high anti-roundup transfer gene plant contains G2-aroA and gat. It has better anti-roundup performance and can be used to culture various high anti-roundup transfer gene plants.

The invention discloses a kit for screening glyphosate N-acetyltransferase antiserum. The invention provides an application of glyphosate N-acetyltransferase in preparation of the kit for preparing and screening glyphosate N-acetyltransferase antiserum. The amino acid sequence of the glyphosate N-acetyltransferase is shown in the table 1), namely amino acid residue of the 114-260th locus from the N-end in the sequence 2 in the sequence table, or table 2), namely the sequence 2 in the sequence table. Experiment shows that by utilizing the method in which the glyphosate N-acetyltransferase antiserum is prepared from prokaryotic expression protein for the first time, the antiserum of GAT (Glyphosate N-acetyltransferase) is successfully obtained and the antiserum can be also applied to specific detection on target protein of transgenic plant.

The invention discloses construction of a three-gene co-expression vector for efficiently synthesizing tetrahydropyrimidine and application of the co-expression vector. The construction method comprises the following steps: sequentially inserting coded L-diaminobutyrate aminotransferase (ectA) gene,coded L-diaminobutyric acid acetyltransferase (ectB) gene and coded tetrahydropyrimidine synthase (ectC) gene in pseudo-bacillus firmus into a transformed plasmid pET-22bNS according to an isocaudarner principle, and constructing the three-gene tandem co-expression vector pET-22bNS-EctA/B/C. When the constructed three-gene co-expression vector is transferred into Escherichia coli BL21 (DE3) and is subjected to IPTG induction, after a recombinant strain whole-cell catalysis reaction for 3 hours, the highest yield of tetrahydropyrimidine is 9.8mg/mL, and the synthetic efficiency is 78.4mg/mL/d and is higher than a synthesizing level in similar researches. The three-gene co-expression vector constructed by the invention has the ability of efficiently synthesizing the tetrahydropyrimidine and has an excellent application value.

This document describes biochemical pathways for producing 6- hydroxyhexanoate methyl ester and hexanoic acid hexyl ester using one or more of a fatty acid O-methyltransferase, an alcohol O-acetyltransferase and a monooxygenase, as well as recombinant hosts expressing one or more of such enzymes. 6-hydroxyhexanoate methyl esters and hexanoic acid hexyl ester can be enzymatically converted to adipic acid, adipate semialdehyde, 6-aminohexanoate, 6-hydroxyhexanoate, hexamethylenediamine, and 1,6-hexanediol.

The invention provides a method for constructing an aspergillus oryzae transgenic system taking phleomycin as a screening marker/GFP (Green fluorescent protein) as a reporter gene. First, codon optimization is performed on an encoded phleomycin resistance gene acetyltransferase blmB; a blmB gene expression cassette is constructed by taking gpdA as a promoter of gene expression and trpc as a terminator; then a pEX1 carrier is transformed into a binary recombinant carrier blmB-pEX1 carrying a blmB gene complete expression nucleus and the fluorescent reporter gene GFP; the recombinant carrier blmB-pEX1 sequentially transforms GV3101 agrobacterium competent cells; then spores of aspergillus oryzae and a bacteria solution of agrobacterium are co-cultured; agrobacterium-mediated aspergillus oryzae gene transfer is performed; after co-culture, screening is performed through a phleomycin resistance screening culture medium; finally, resistant transformants are picked for expanded culture, anda fluorescent microscope is used to observe the successful expression of the GFP. The false positive rate of the transformants is effectively reduced, the screening workload is reduced, and the problem of insufficient resistance screening markers is solved.

The invention discloses recombinant escherichia coli for producing melatonin, and a construction method and application of the recombinant escherichia coli. The invention first discloses the recombinant escherichia coli, and compared with receptor escherichia coli, the recombinant escherichia coli has the advantages that the expression amount of a gene of a melatonin biosynthesis-related protein is increased, and/or the content of the melatonin biosynthesis-related protein is increased, and/or activity of the melatonin biosynthesis-related protein is increased, wherein the melatonin is selected from at least one of phenylalanine hydroxylase, dihydromonophosphate reductase, 4alpha-hydroxytetrahydrobiopterin dehydratase, aspartate transaminase family protein, N-acetyltransferase and caffeicacid-O-methyltransferase.and further discloses a construction method and application of the recombinant escherichia coli. The recombinant escherichia coli strain for producing melatonin has melatoninsynthesis efficiency is obviously higher than other existing strains, and has great significance for industrial production and large-scale application of melatonin.

The invention discloses 432 novel acetylation sites identified in signal transduction proteins and pathways underlying human protein acetylation signaling pathways, and provides acetylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these acetylated sites/proteins, as well as methods of using the reagents for such purpose. Among the acetylation sites identified are sites occurring in the following protein types: Acetyltransferases, Adaptor/Scaffold proteins, Actin binding proteins, Adhesion proteins, Apoptosis proteins, Calcium-binding proteins, Cell Cycle Regulation proteins, Cell Surface proteins, DNA binding proteins, DNA replication proteins, Channel proteins, Chaperone proteins, Cellular Metabolism enzymes, Cytoskeletal proteins, DNA repair proteins, Endoplasmic reticulum proteins, Enzyme proteins, G protein and GTPase Activating proteins, Guanine Nucleotide Exchange Factors, Helicase proteins, Isomerase proteins, Extracelluar matrix proteins, Hydrolases, Ligase proteins, Lipid kinases, Inhibtor proteins, Lipid Binding proteins and Lyases.

The invention relates to an indolinone derivative having excellent lecithin-cholesterol acetyltransferase (LCAT), preferably reversible LCAT activation effect as well as its pharmaceutically acceptable salt. The compound comprises a compound shown in a formula I or its pharmaceutically acceptable salt. The compound has excellent excitement effect on LCAT, has obvious increase effect for HDL level, and is used for preventing and treating cardio cerebrovascular diseases such as coronary heart disease and arteriosclerosis.

The invention relates to a naphthalimide derivative using an ester group for modifying an amonafide naphthalene ring and a preparation method and application of the naphthalimide derivative. The naphthalimide derivative using the ester group for modifying the amonafide naphthalene ring and the preparation method and application of the naphthalimide derivative have the advantages that a carboxy group is introduced into the fourth position of the amonafide naphthalene ring to be coupled with fatty alcohol in different chain length, a 3-position amino group in an amonafide parent ring is removed, and accordingly acetylation of in-vivo acetyltransferase on the amino group on an amonafide ring is stopped, the possibility that N-acetylated amonafide causes unpredictable toxicity in vivo is avoided, and excellent in-vitro tumor cell inhibiting capability is achieved to obtain a new high-efficiency low-toxicity medicine.

The invention discloses a full-biosynthesis method of pimelic acid, and belongs to the field of bioengineering. Escherichia coli BL21(DE3) with a lactate dehydrogenase gene (ldhA), an acetyl-CoA acetyltransferase gene (atoB) and a succinyl-CoA synthetase alpha subunit gene (sucD) knocked out is used as a host, genes of beta-ketothiolase, 3-hydroxyacyl-coenzyme A dehydrogenase, 3-hydroxyadipyl dehydrogenase, 5-carboxyl-2-pentenoyl coenzyme A reductase and adipyl coenzyme A synthetase from thermopsis fusca are overexpressed in modules, and the yield of pimelic acid can reach 252.60mg.L < -1 > through fermentation optimization of added inorganic salt in a culture medium. A new route is provided for synthesis of pimelic acid in the Escherichia coli host, and a new idea is provided for industrial fermentation production of pimelic acid.

The invention belongs to the technical field of bioengineering, and particularly relates to an ester catechin synthase, a coding gene and application of the ester catechin synthase. A recombinant protein is obtained by constructing a recombinant engineered bacteria, with the help of an enzyme reaction method, a recombinant protein of a genetically engineered bacteria is used as a catalytic enzyme,and genes, encoding the serine carboxypeptidase acylase CsSCPL11-IA, CsSCPL13-IA and CsSCPL14-IA, in tea plants are verified. Three acetyltransferase genes that can catalyze the synthesis of ester catechins and the application values in the breeding of excellent tea varieties and tea processing are described for the first time.