32 results about "Guanosine triphosphate" patented technology

A process is disclosed for the production of cyclic di-guanosine monophosphate (c-di-GMP) without the use of protecting groups by means of an enzymatic synthesis. The process comprises the coupling of two guanosine triphosphate (GTP) molecules so as to form a c-di-GMP molecule. This is done under the influence of a mutant diguanylate cyclase (DGC) comprising the amino acid sequence V153M154G155G156. It has been found that the DGC is obtainable from inclusion bodies, and therewith can be made available in amounts sufficient to improve the c-di-GMP synthesis. Particularly, the latter synthesis can be conducted in a one-pot method starting from commercially available bulk chemicals and allows upscaling to a commercial production scale.

The invention relates to a reagent for detection of biological activity of glucagon-like peptide-1 (GLP-1). Each 100 microliters of the reagent mainly comprises 20 micrograms of a gene recombinant GLP-1 acceptor, a dipeptidyl peptidase-4 inhibitor of which the weight is 1% of the weight of a blood plasma sample, 480 micrograms of triphosadenine, 10 micrograms of guanosine triphosphate, 0.5 milligrams of 3-isobutyl-1-methylxanthine and 100 microliters of a membrane reaction solution. The GLP-1 acceptor is obtained by genetic recombination, is convenient for purification and reduces batch difference of the reagent. The GLP-1 acceptor is used as a probe for capturing GLP-1 so that detection sensitivity is improved, biological activity can be directly determined, sensitivity is high, and practicality is strong.

The invention relates to a reagent for detection of biological activity of glucagon-like peptide-1 (GLP-1). The reagent added into each 15 microliters of a detection sample comprises: 1, 35-45 microliters of a membrane reaction solution, 2, 15-25 microliters of a GLP-1 acceptor, 3, 10-20 microliters of a GLP-1 standard substance, 4, 5-15 microliters of sitgliptin phosphate, 5, 1-10 microliters of triphosadenine, 6, 1-10 microliters of guanosine triphosphate, 7, 5-15 microliters of 3-isobutyl-1-methylxanthine, 8, 1-10 microliters of bovine serum albumin, 9, 50-150 microliters of a biotin-labeled mouse anti-human cAMP monoclonal antibody, 10, a microwell plate coated with a goat anti-human cAMP polyclonal antibody, 11, 50-150 microliters of an avidin-labeled horseradish peroxidase, 12, 300-400 microliters of a PBS washing liquor, 13, 50-150 microliters of a chromogenic substrate, and 14, 50-150 microliters of a stop buffer. The GLP-1 acceptor is produced by genetic recombination, is convenient for purification and can reduce the batch difference of the reagent. The GLP-1 acceptor is used as a probe for capturing GLP-1 so that detection sensitivity is improved, biological activity can be directly determined, sensitivity is high, and practicality is strong.

The present invention provides a fermentation method for increasing the yield of riboflavin, the gluconic acid metal complex is added into the fermentation culture medium, fermentation nutrients are fed, the functions of carbohydrates can be diversified, absorption of thalli to metal elements is optimized, and the toxic and side effects of the metal elements to the thalli are reduced; along with the proceeding of fermentation, the nutritional ingredients in the fermentation tank are gradually reduced, the vitality of thalli is gradually weakened, and fed-batch fermentation nutrients can supplement the nutritional ingredients required by the thalli in the fermentation process and maintain the growth vitality of the thalli; due to the addition of citric acid, the EMP pathway is inhibited, and the HMP pathway is enhanced; glutamine enhances the generation of riboflavin precursor guanosine triphosphate, and is beneficial to the accumulation of riboflavin; and the bacillus subtilis is used as a growth coupling type strain for producing riboflavin, the quantity of thalli influences the yield of riboflavin to a great extent, and the fed-batch of KH2PO4 can keep the thalli in a good growth state, so that the yield of riboflavin is increased.

The invention discloses a small G-protein Rop family protein, a coding gene and an application of the hevea brasiliensis latex small G-protein Rop family protein. The protein is a protein having one of the following amino acid residue sequences: 1) protein composed of an amino acid residue sequence of SEQ ID No.1 in a sequence table; 2) SEQ ID No.1 derived protein obtained by subjecting the amino acid residue sequence of SEQ ID No.1 in the sequence table to substitution and / or deletion and / or addition of one or more amino acid residues and having GTP (guanosine triphosphate) binding function. The protein has GTP binding activity, plays a certain role in basic disease-resistant mechanism and stress resistance of plants, and has the characteristics of control for secondary xylem and phloem development of woods. The protein can improve the resistance of transgenic crop or woods against adverse situation, so as to improve the yield.

The invention provides methods to inhibit the replication of a flavivirus, methods of inhibiting the guanosine triphosphate (GTP)-binding and guanylyltransferase activity of a flavivirus RNA capping enzyme, and methods of treating a subject infected with a flavivirus. The methods can include contacting a flavivirus with an effective amount of a thioxothiazolidine compound described herein, or a derivative thereof, such as a compound of Formula (I).

The invention discloses a method for testing BNP (Brain Natriuretic Peptide) activity by using a GCA stable transfection cell line and establishes a novel BNP bioactivity testing method. GCA which is a main acceptor of BNP is an adenylate cyclase precursor and can catalyze GTP (guanosine triphosphate) to generate cGMP after being activated by the BNP and then exerts a biological effect. Wield 293 cell has low GCA expression level and low reactivity to BNP. In the method disclosed by the invention, a stable cell line 293GCAC3 with BNP acceptor (GCA) overexpression level is structured by transgenosis, the content change of cGMP secreted into cell supernatant by the 293GCAC3 stimulated by the BNP is detected to quantify the activity of the BNP. The method comprises the following specific testing steps of: laying cells on a 96-pore plate and culturing for 16-18 hours; adding the BNP for stimulating for 90 minutes; and sucking cell supernatant and detecting the content of cGMP with competitive ELISA (enzyme-linked immuno sorbent assay). The method is simple to operate, has sensitive response and small variability and is significant for the quality control and clinical application of the BNP.

The invention discloses a detection kit based on an aptamer. The detection kit comprises stem-loop structure nucleic acid H1, DNA (deoxyribonucleic acid) 1, DNA2 and DNA3, the stem-loop structure nucleic acid comprises the aptamer divided into two sections, the aptamer serves as a molecular recognition element, specific binding of molecules to be detected is achieved, catalytic activity of DNA enzymes is activated, substrate chains are circularly cut, high fluorescent detection signals are generated, and high-sensitivity detection is achieved. Detection limit of the detection kit for ATP (adenosine triphosphate) is 0.2 nanometer, detected linear range is wide and is 1 nanometer to 10 micrometers, a detection system has excellent specificity, common analogues comprising CTP (cytidine triphosphate), GTP (guanosine triphosphate) and UTP (uridine triphosphate) do not interfere detection results, separation and purification processes are omitted in the detecting process, detection can be achieved by simple blending, the detection kit has the advantages of simplicity in operation, low cost, quick response and the like, and the detection kit can be used for rapidly detecting and analyzingthe content of the ATP in environments or food samples.

The invention belongs to the technical field of scale production of candidate medicines for treating glucose-lipid metabolism disorder diseases. In order to solve the problems that existing S-AMP is difficultly produced, the price of the S-AMP is expensive, and raw material sources of the druggability of the S-AMP is difficulty ensured, the invention provides a preparation method of adenylosuccinic acid or adenylosuccinate. PhAdSS of which the N terminal is provided with an His-tag sequence is expressed by utilizing a pET-28-a carrier; with the PhAdSS as a catalyst, inosinic acid IMP, L-aspartic acid and guanosine triphosphate (GTP) are catalyzed and subjected to reaction to synthesize S-AMP; after the reaction is carried out, components in a product are analyzed by adopting silica gel thin-layer chromatography, and separation and purification on the S-AMP are carried out by adopting silica gel coluumn chromatography. The content of the s-AMP in a synthetic product is 61 percent, and arecycling ratio is about 20 percent. A preparation process is simple, the cost is low, and a basis is laid for building an industrialized S-AMP synthetic process; monopoly sale on S-AMP by foreign sellers is expected to be broken through, and the S-AMP druggability research cost can be greatly reduced.

The invention relates to application of guanosine triphosphate cyclohydrolase 1 (GCH1) and a drug. It is found that GCH1 overexpression has the prevention and treatment effect on radiation injuries ofhuman lung cells and animal model lung tissues, and can enhance oxidation resistance of the cells and cell proliferation capacity after radiation. A GCH1 overexpression adenovirus or the drug containing the GCH1 overexpression adenovirus can be used for preventing and treating human acute lung radiation injuries, and has the important significance and application value in coping with nuclear andradiation accidents and nuclear emergencies, reducing the lung radiation injury effects on tumor radiation patients and the like.