42 results about "Cytidine diphosphate" patented technology

The present disclosure relates to nutritional compositions comprising a neurologic component, wherein, the neurologic component may promote brain and nervous system development and further provide neurological protection and repair. The neurologic component may include phosphatidylethanolamine, sphingomyelin, cytidine diphosphate-choline, ceramide, uridine, at least one ganglioside, and mixtures thereof. The disclosure further relates to methods of promoting brain and nervous system health by providing said nutritional compositions to target subjects, which includes pediatric subjects.

A substantially pure culture of a microorganism having enzyme activities of pyrG and CCT and carrying a recombinant DNA composed of a DNA fragment containing genes encoding pryG and CCT and a vector.

The invention relates to a fluorescent probe, which belongs to the technical field of fine chemical engineering, and discloses a rhodamine fluorescent probe with a pseudo nucleic acid base as a recognition site and preparation thereof and application to a nucleotide image. The ring opening of rhodamine lactone is induced between amino naphthyridine and nucleic acid base through a base analogue complementary hydrogen bond effect, and the selective response of specific nucleotide is realized by using the weakness of electrostatic interaction between imine positive ions generated by a ring opening product and phosphoric acid link negative ions with proper length. The probe shows good selectivity to cytidine diphosphate (CDP) and adenosine triphosphoric acid (ATP). Hela cell copolymerization focal imaging experiments show that the probe can produce a sensitive imaging effect on the nucleotide in the cell, the cell state is good after and before imaging, and the probe has small cell toxicity. Since the probe has good bioavailability and a high-sensitivity imaging effect, the probe is widely applied to medical fluorescent imaging.

The invention discloses a method for separating purified cytidine diphosphate choline by hydrophobic chromatography. The method comprises the following steps of: (1), performing enzyme inactivation, centrifugation and ultra-filtration of a cytidine diphosphate choline conversion liquid; (2), after adjusting the pH of the pre-processed cytidine diphosphate choline conversion liquid to between 2.0 and 14.0, adding an inorganic salt to prepare column-loading liquid which contains 0.01 to 5mol / L of inorganic salt and 0.1 to 40g / L of cytidine diphosphate choline with the cytidine diphosphate choline conversion liquid, absorbing the column-loading liquid with a hydrophobic chromatography resin and eluting with pure water of which the pH value is 2.0 to 14.0; and (3), nano-filtering, desalinating and concentrating the eluate, and obtaining the cytidine diphosphate choline through crystallization. In the method, the separation process is simple; through regeneration, the resin after elution can be repeatedly used; the separation cost is low; the product is easily crystallized; and the CDP-choline product with high purity and high yield can be obtained.

The present disclosure relates to nutritional compositions comprising a neurologic component, wherein, the neurologic component may promote brain and nervous system development and further provide neurological protection and repair. The neurologic component may include phosphatidylethanolamine, sphingomyelin, cytidine diphosphate-choline, ceramide, uridine, at least one ganglioside, and mixtures thereof. The disclosure further relates to methods of promoting brain and nervous system health by providing said nutritional compositions to target subjects, which includes pediatric subjects.

The invention relates to a manufacturing method of disodium cytidine triphosphate, which comprises the following steps: proceeding enzymatic conversation with CMP material; proceeding ion exchange column chromatography; separating and purifying. The enzymatic conversation reaction principle is that: brewers' yeast gets the needed enzyme liquid of the enzymatic conversation reaction; brewers' yeast produces a plurality of biological enzyme as sugar glycolysis enzyme and phosphorylase with high sugar glycolysis ability; the biological energy produced by glycolysis glucose with sugar glycolysis enzyme is transferred to cytidine monophosphate by ATP; cytidine monophosphate and phosphate radical combines to form cytidine triphosphate with the effect of phosphorylase. The method shapes low cost, simple operation, high production, high product purity by optimization, which comprises the following steps: getting enzyme liquid; proceeding enzymatic conversation by adding the main material cytidine monophosphate, monobasic potassium phosphate of proper quantity and glucose; getting product by deposition, ion exchange column chromatography, segregation, purification, refining and vacuum drying.

The invention relates to a process for preparing cytidine diphosphate choline. According to the method, denovo synthesis of pyrimidine is utilized as a basic reaction and choline materials are subjected to a reaction at the reaction temperature ranging from 20 DEG C to 38 DEG C for 10 to 72 hours with pH controlled at 5 to 7.5 in the presence of culture solution of microorganism A or further in the presence of culture solution of microorganism B or treatment so as to accumulatively produce cytidine diphosphate choline. Simple starting materials of carbon and nitrogen sources are utilized, in culture solution containing the choline materials, precursor of the cytidine diphosphate choline, namely cytidine monophosphate is synthesized by means of metabolic process, and cytidine diphosphate choline is cumulatively produced in the reaction solution. According to the method, expensive cytidine monophosphate or the precursor materials are avoided so that the preparing cost of the cytidine diphosphate choline is greatly reduced.

The invention discloses a preparation method of a gene DNA (Deoxyribose Nucleic Acid) sequence capture probe. According to the technical scheme, the method comprises the following steps: 1) designing two partially complementary primers and carrying out annealing treatment so as to form a partially double-chain structured split joint, and carrying out a coupled reaction between the split joint and an ordinary PCR (Polymerase Chain Reaction) amplification product (A1) of a target gene so as to obtain a target gene (A2) two ends of which are provided with joints; 2) designing corresponding primers according to the sequences of the joints and amplifying the target gene (A2) so as to obtain an amplification product (A3) with a T7 promoter; and 3) quantifying the product (A3), and adding NTP (Nucleotide Triphosphate) with biotin labels (wherein biotins are labeled on the NTP is labeled with biotins but ATP (Adenosine Triphosphate), CTP (Cytidine Triphosphate) and DTP (Deoxyadenosine Triphosphate) are not labeled with biotins) into the product (A3) under a certain condition so as to carry out an in-vitro transcription reaction, thereby finally obtaining an RNA (Ribose Nucleic Acid) probe with biotin labels (gene DNA sequence capture probe).

The invention relates to an artemisia apiacea4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol synthase coding sequence in the technical field of gene engineering. The sequence is a sequence generated after one or a plurality of codons in nucleotide are replaced by degenerate codons of amino acid with same coding, as shown in 18-position to 926-position of SEQ ID NO: 3 or shown in 18-position to926-position of SEQ ID NO: 3. The method for improving the content of plant artemisinin by using the coding sequence includes the steps of: connecting the coding sequence to a plant expression regulatory sequence and constructing a plant expression carrier containing the coding sequence; transferring the expression carrier in step 1 to agrobacterium rhizogenes, then transferring agrobacterium rhizogenes to artemisia apiacea; through antibiotic screening, obtaining a transformation cell containing the coding sequence and regenerating a transgenosis plant. The invention relates to a polypeptide, and the sequence is shown in SEQ ID NO: 4. The coding sequence can improve the content of artemisinin in artemisia apiacea and the content of isoprene compound of other plants.

The invention discloses a preparation method of uridine-5'-monophosphate disodium. The method comprises the following steps: (1) mixing cytidine monophosphate (CMP) or sodium salt, sodium nitrite and deionized water, dropwise adding acid and / or acid anhydride, continuously reacting for 0-6 hours after completing dropwise adding to obtain reaction solution 1; (2) if residual quantity of CMP is lower than 1%, adjusting the pH value of the reaction solution 1 to 6.4-7.2 to obtain reaction solution 2; (3) mixing the reaction solution 2 and 95v / v% of ethyl alcohol, crystallizing and filtering to obtain a uridine-5'-monophosphate disodium crude product; (4) mixing the uridine-5'-monophosphate disodium crude product and water, and adjusting the pH value to 7.0-8.5 to obtain reaction solution 3; and (5) mixing the reaction solution 3 and an organic solvent, recrystalizing and filtering to obtain the uridine-5'-monophosphate disodium.

The invention discloses an amphipathic Eu (III) complex as well as a preparation method and application thereof in detection for cytidine triphosphate. The structural formula of the complex is as shown in the specification. The amphipathic Eu (III) complex is formed by matching a ligand with Eu (III), wherein a compound which takes octadecyl as hydrophobic tail part and takes 1,4,7,10-tetraazacyclododecane as a hydrophobic head part serves as the ligand. The preparation method is simple and the reaction condition is mild. The amphipathic Eu (III) complex provided by the invention has the characteristics of long fluorescent lifetime, large stokes shift and unsaturated coordination; a super-molecule sensing interface is assembled and formed in a water phase; the energy donor alpha-naphthyl phosphate sodium is introduced into the system and the amphipathic Eu (III) complex serves as an energy receptor for receiving energy and emitting Eu (III) characteristic light; when the cytidine triphosphate is contained in a to-be-detected sample, the alpha-naphthyl phosphate sodium on the surface of the self-assembly body is replaced by the cytidine triphosphate, so that the fluorescence is quenched and the selective identification for the cytidine triphosphate is realized.

The invention relates to a medicinal preparation of a compound, and especially relates to a cytidine disodium triphosphate injection liquid, and a preparation method thereof. The preparation comprises cytidine disodium triphosphate, propylene glycol, and injection water. The preparation method comprises steps that: a small amount of injection water (with a temperature cooled to below 20 DEG C), propylene glycol and cytidine disodium triphosphate are fetched; cytidine disodium triphosphate is added to injection water, and is completely dissolved; propylene glycol is added to the solution, and is well-mixed by stirring; a 4% sodium hydroxide solution is slowly dropped into the solution, until the pH value of the solution is 8.6-.8.9; injection-use active carbon which takes 0.1% (g / ml) of the solution by volume is added to the solution, and the solution is stirred for 15 minutes; the solution is filtered, and injection water is added until a full dose is reached; and the solution is then processed through filtering, filling, sealing, lamp inspecting, and packaging. The injection liquid provided by the invention has advantages of simple formulation, simple technology, high production efficiency, strong medicine liquid stability, and the like.

The invention discloses a genetic engineering preparation method of cytidine triphosphate. In the method, a culture or a treatment substance of single genetic engineering strain is used as an enzyme source; during catalysis, ammonium chloride, saratin and the like are used for reaction, so that the cytidine triphosphate is generated in a reaction solution and accumulated and is extracted from the reaction solution. The invention has the benefits that: (1) the reaction is easier to control due to the adoption of the microbial catalytic reaction ; (2) the cost of a substrate is lower and the cytidine triphosphate can be produced at low cost; and (3) the reaction is quick and the conversion rate is higher. The genetic engineering preparation method can be widely applied to preparation of the cytidine triphosphat.

The invention belongs to a preparation method of poly i-c and belongs to the technical field of biomedicine. The preparation method includes: utilizing inosine diphosphate to prepare polyinosinic acid, and using cytidine diphosphate to prepare polycytidylic acid; utilizing the prepared polyinosinic acid to react with the polycytidylic acid to obtain a high-purity poly i-c product. Poly i-c is prepared through self-developed high-purity polyinosinic acid and polycytidylic acid, so that prepared poly i-c is high in purity, and treatment difficulty in the process of preparing is lowered.

The invention discloses a preparation method of a gene DNA (Deoxyribose Nucleic Acid) sequence capture probe. According to the technical scheme, the method comprises the following steps: 1) designing two partially complementary primers and carrying out annealing treatment so as to form a partially double-chain structured split joint, and carrying out a coupled reaction between the split joint and an ordinary PCR (Polymerase Chain Reaction) amplification product (A1) of a target gene so as to obtain a target gene (A2) two ends of which are provided with joints; 2) designing corresponding primers according to the sequences of the joints and amplifying the target gene (A2) so as to obtain an amplification product (A3) with a T7 promoter; and 3) quantifying the product (A3), and adding NTP (Nucleotide Triphosphate) with biotin labels (wherein biotins are labeled on the NTP is labeled with biotins but ATP (Adenosine Triphosphate), CTP (Cytidine Triphosphate) and DTP (Deoxyadenosine Triphosphate) are not labeled with biotins) into the product (A3) under a certain condition so as to carry out an in-vitro transcription reaction, thereby finally obtaining an RNA (Ribose Nucleic Acid) probe with biotin labels (gene DNA sequence capture probe).

The invention relates to an artemisia apiacea4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol kinase coding sequence in the technical field of gene engineering. The sequence is a sequence generated after one or a plurality of codons in nucleotide are replaced by degenerate codons of amino acid with same coding, as shown in 15-pisition to 1214-position of SEQ ID NO: 3 or shown in 15-position-1214-position of SEQ ID NO: 3. The method for improving the content of plant artemisinin by using the coding sequence includes the steps of: connecting the coding sequence to a plant expression regulatory sequence and constructing a plant expression carrier containing the artemisia apiacea4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol kinase coding sequence; transferring the expression carrier in step 1 to agrobacterium rhizogenes, then transferring agrobacterium rhizogenes to artemisia apiacea; through antibiotic screening, obtaining a transformation cell containing the artemisia apiacea4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol kinase coding sequence and regenerating a transgenosis plant. The coding sequence can improve the content of artemisinin in artemisia apiacea and the content of isoprene compound of other plants.

The invention relates to a manufacturing method of disodium cytidine triphosphate, which comprises the following steps: proceeding enzymatic conversation with CMP material; proceeding ion exchange column chromatography; separating and purifying. The enzymatic conversation reaction principle is that: brewers' yeast gets the needed enzyme liquid of the enzymatic conversation reaction; brewers' yeast produces a plurality of biological enzyme as sugar glycolysis enzyme and phosphorylase with high sugar glycolysis ability; the biological energy produced by glycolysis glucose with sugar glycolysis enzyme is transferred to cytidine monophosphate by ATP; cytidine monophosphate and phosphate radical combines to form cytidine triphosphate with the effect of phosphorylase. The method shapes low cost, simple operation, high production, high product purity by optimization, which comprises the following steps: getting enzyme liquid; proceeding enzymatic conversation by adding the main material cytidine monophosphate, monobasic potassium phosphate of proper quantity and glucose; getting product by deposition, ion exchange column chromatography, segregation, purification, refining and vacuum drying.

The invention discloses a preparation method of powdery cytidine triphosphate or a sodium salt of cytidine triphosphate. The preparation method comprises the following steps: mixing and precipitating an acid solution containing cytidine triphosphate or the sodium salt of cytidine triphosphate with methanol of which the volume is 5-20 times of that of the acid solution so as to obtain powdery cytidine triphosphate or the sodium salt of powdery cytidine triphosphate, or the preparation method comprises the following steps: (1) mixing the acid solution containing cytidine triphosphate or the sodium salt of cytidine triphosphate with a first organic solvent of which the volume is 2-20 times of that of the acid solution so as to obtain a first precipitate; (2) mixing the first precipitate with ethanediol or glycerin of which the volume is 2-20 times of that of the first precipitate so as to obtain a dissolved solution; (3) mixing and precipitating the dissolved solution with a second organic solvent of which the volume is 2-20 times of that of the dissolved solution so as to obtain powdery cytidine triphosphate or the sodium salt of powdery cytidine triphosphate.

The invention discloses a PLIspE gene and a coded protein and application thereof. The gene is cloned from Paeonia lactiflora by constructing a cDNA library for the first time, so a gap in separation and cloning of a 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase gene from the traditional Chinese medicine--Paeonia lactiflora is filled in. The PLIspE gene provided by the invention has a nucleotide sequence shown in SEQ ID NO. 1, or a homologous sequence obtained through addition, substitution, insertion or deletion of one or a plurality of nucleotides of the nucleotide sequence shown in SEQ ID NO. 1, or an allele of the gene or a nucleotide sequence derived from the allele. The protein coded by the gene has an amino acid sequence shown in SEQ ID NO. 2 or a homologous sequence obtained through addition, substitution, insertion or deletion of one or a plurality of amino acids of the amino acid sequence shown in SEQ ID NO. 2. The PLIspE gene can increase the content of metabolites of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol in Paeonia lactiflora through biotechnology and has good application prospects in aspects like quality improvement of the medicinal material Paeonia lactiflora and synthetic biology of active components of Paeonia lactiflora.

The invention relates to a combination of cytidine disodium triphosphate and arginine, and the preparation method thereof; the freeze-dried powder adopts arginine as the pH regulator, has good stability, long storage time, and no local irritation and pain.

The invention discloses a PLIspE gene and a coded protein and application thereof. The gene is cloned from Paeonia lactiflora by constructing a cDNA library for the first time, so a gap in separation and cloning of a 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase gene from the traditional Chinese medicine--Paeonia lactiflora is filled in. The PLIspE gene provided by the invention has a nucleotide sequence shown in SEQ ID NO. 1, or a homologous sequence obtained through addition, substitution, insertion or deletion of one or a plurality of nucleotides of the nucleotide sequence shown in SEQ ID NO. 1, or an allele of the gene or a nucleotide sequence derived from the allele. The protein coded by the gene has an amino acid sequence shown in SEQ ID NO. 2 or a homologous sequence obtained through addition, substitution, insertion or deletion of one or a plurality of amino acids of the amino acid sequence shown in SEQ ID NO. 2. The PLIspE gene can increase the content of metabolites of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol in Paeonia lactiflora through biotechnology and has good application prospects in aspects like quality improvement of the medicinal material Paeonia lactiflora and synthetic biology of active components of Paeonia lactiflora.

The present application belongs to the technical field of bioengineering, and particularly relates to an engineered bacterium for producing cytidine diphosphate choline as well as a construction method and application thereof. The present application takes Bacillus subtilis BSC1-4 as a starting bacterium, blocks the conversion of choline to glycine betaine to reduce unnecessary loss of substrates in the fermentation process, improves the absorption of choline, and enables the substrates to continuously generate cytidine diphosphate choline in the direction of generating cytidine diphosphate choline. The construction method of the engineered bacterium is specifically as follows: the key genes of the choline degradation pathway in the genome of Bacillus subtilis are knocked out gbsA and gbsB , the transporter protein OpuD is overexpressed, and finally the transcriptional repressor genes opcR and gbsR are knocked out. Experimental results show that the engineered bacterium constructed in the present application has a cytidine diphosphate choline yield of 1181.6±11.5 mg / L after 24 h of fermentation in a shake flask.

Compositions and method are therefore disclosed for treating ARDS. In particular, disclosed a composition that contains one, two, or more cytidine diphosphate (CDP)-conjugated precursors selected fromthe group consisting of CDP-choline, CDP-ethanolamine, and CDP-diacylglycerol (CDP-DAG) in a pharmaceutically acceptable carrier for use in treating ARDS.

The invention relates to CTP (Cytidine Triphosphate) synthetase from Bailing producing strain cordyceps sinensis hirsutella sinensis participated in synthesis of metabolic cytidine triphosphate from uridine triphosphate, a gene for coding the synthetase and application of the CTP synthetase; the CTP synthetase has an amino acid sequence represented by SEQ ID No.2 and with the homology of more than 90%; the coding gene has a nucleotide sequence represented by SEQ ID No.4 and with the homology of more than 90%; a metabolic pathway of synthesis of cytidine triphosphate by uridine triphosphate is researched in a detailed manner in principle; cloned DNA of the nucleotide sequence provided by the invention can be transferred into engineering bacteria through transduction, conversion and conjugal transfer methods; by adjusting expression of a biosynthetic gene of cytidine triphosphate, the high expressivity of host cytidine triphosphate is given; an effective way is provided for increasing the yield of cytidine triphosphate; and therefore, the CTP synthetase has great application prospect.

The application discloses a banana gene MaRBL1 for improving disease resistance of plants, a method and application, wherein the banana gene MaRBL1 nucleotide sequence is shown as SEQ ID NO. 1; the gene encodes cytidine diphosphate diacylglycerol synthetase, is a key enzyme for phospholipid biosynthesis, and participates in synthesis of phosphatidylinositol and derivatives phosphatidylinositol 4, 5-diphosphate; a core functional domain of the gene is a CTP_transf_1 domain, retains catalytic activity of CDP-DAG synthetase, participates in phospholipid metabolism and conduction of disease resistance signals, and the application provides a new strategy for cultivating high-resistance banana varieties.

The invention belongs to a preparation method of poly i-c and belongs to the technical field of biomedicine. The preparation method includes: utilizing inosine diphosphate to prepare polyinosinic acid, and using cytidine diphosphate to prepare polycytidylic acid; utilizing the prepared polyinosinic acid to react with the polycytidylic acid to obtain a high-purity poly i-c product. Poly i-c is prepared through self-developed high-purity polyinosinic acid and polycytidylic acid, so that prepared poly i-c is high in purity, and treatment difficulty in the process of preparing is lowered.

The invention relates to a CTP (Cytidine Triphosphate) synthetase which is from Bailing production bacterium cordyceps sinensis hirsutella sinensis and takes part in synthesis of metabolic cytidine triphosphate from uridine triphosphate, a gene encoding the synthetase and an application of the synthetase. The CTP synthetase has more than 90% of homology of an amino acid sequence represented by SEQ ID No.1 or SEQ ID No. 2, and the encoding gene of the synthetase has more than 0% of homology of a nucleotide sequence represented by SEQ ID No. 3 or SEQ ID No.4. According to the invention, a metabolic pathway for synthesizing the cytidine triphosphate from the uridine triphosphate is deeply studied in principle. Cloned DNA (Deoxyribonucleic Acid) comprising the nucleotide sequence provided by the invention can be transferred into engineering bacteria by transduction, conversion and transconjunction, and the expression of a biological synthesis gene of the cytidine triphosphate is regulated to ensure high expressivity of the host cytidine triphosphate, so that an effective way for enlarging the output of the cytidine triphosphate is provided, and the prospect in application is great.

The application discloses a method for improving survival rate of probiotics based on cell membrane regulation, and the method comprises the following steps: adding lipid components during the culture of probiotics and / or adding the lipid components as a protective agent after the culture of probiotics; the lipid components added during the culture of probiotics comprise cytidine diphosphate diacylglycerol and / or stearic acid; and the lipid components added as a protective agent after the culture of probiotics comprise cardiolipin and / or oleic acid. The method breaks through the research scope of traditional optimization of freeze-drying process (such as protective agent and parameter), and through the establishment of lipidomics means and a screening method of anti-resistance related lipid molecules, key lipid molecules directly related to resistance are screened out; through the dual mode of supplementing key lipid components and precursor substances, the strain resistance is improved from the metabolic level, and the freeze-drying resistance and survival rate of the strain are significantly improved.

The invention discloses a genetic engineering preparation method of cytidine triphosphate. In the method, a culture or a treatment substance of single genetic engineering strain is used as an enzyme source; during catalysis, ammonium chloride, saratin and the like are used for reaction, so that the cytidine triphosphate is generated in a reaction solution and accumulated and is extracted from the reaction solution. The invention has the benefits that: (1) the reaction is easier to control due to the adoption of the microbial catalytic reaction ; (2) the cost of a substrate is lower and the cytidine triphosphate can be produced at low cost; and (3) the reaction is quick and the conversion rate is higher. The genetic engineering preparation method can be widely applied to preparation of the cytidine triphosphat.

The invention discloses a method for synthesizing cytidine diphosphate, and belongs to the field of nucleoside synthesis in organic chemistry. The method comprises the following reaction steps that cytidine monophosphate and dichlorophosphoryl morpholine serve as raw materials, condensation is conducted under the base catalysis effect, then a crude product cytidine diphosphate is obtained through acyl chloride hydrolysis, recrystallization is conducted through an alcohol / water mixed system, and a finished cytidine diphosphate product with the HPLC purity larger than 95% is obtained. The whole reaction only comprises two steps, reaction operation is simple, and the total yield reaches 70% or above.