33 results about "Cytosine nucleotide" patented technology

The present invention provides for novel methods for regulating and detecting the cytosine methylation status of DNA. The invention is based upon identification of a novel and surprising catalytic activity for the family of TET proteins, namely TET1, TET2, TET3, and CXXC4. The novel activity is related to the enzymes being capable of converting the cytosine nucleotide 5-methylcytosine into 5-hydroxymethylcytosine by hydroxylation.

The present invention provides a means to modulate gene expression in vivo in a manner that avoids problems associated with CRISPR endogenous protein knock-out or knock-in strategies and strategies that provide for correction, or alteration, of single nucleotides. The invention includes inserting into the genome a nucleotide encoding a heterobifunctional compound targeting protein (dTAG) in-framewith the nucleotide sequence of a gene encoding an endogenously expressed protein of interest which, upon expression, produces an endogenous protein-dTAG hybrid protein. This allows for targeted protein degradation of the dTAG and the fused endogenous protein using a heterobifunctional compound.

The invention discloses a quality control method for ribonucleic acid II for injection. The method comprises the following steps that the nucleic acid enzyme hydrolysis solution of a substance to be measured is subjected to high-performance liquid chromatogram analysis, an adopted chromatographic column is an Agilent ZORBAX SB-AQC18 chromatographic column, and a flowing phase is a mixture of a formic acid solution and an acetonitrile solution; after the analysis, if the substance to be measured is determined to contain five substances as follows: cytidylate, uridine monophosphate, guanine nucleotide, guanosine and adenosine, the substance to be measured is the ribonucleic acid II for injection or is the ribonucleic acid II for injection as a candidate; and if not, the substance to be measured is not the ribonucleic acid II for injection or is not the ribonucleic acid II for injection as the candidate. A high-performance liquid chromatographic technique is utilized, and the strong-specificity quality control method for the ribonucleic acid II for injection is established. The method has important meanings on increasing the technological content of the medicine, increasing the safety and effectiveness, reducing the cost, enlarging the production scale, increasing the market occupancy, and going forward to the international market.

The invention discloses a chemical labeling and LC-MS combined method, and applications thereof in nucleotide analysis. According to the chemical labeling and LC-MS combined method, N,N-dimethyl-p-phenylenediamine is used for chemical labeling of 10 monophosphate nucleotides containing two methylation modified nucleotides, so that the retention behavior of the modified nucleotides in reversed phase liquid chromatography is improved obviously, and detection sensitivity of the modified nucleotides in mass spectrometric detection is increased. The chemical labeling and LC-MS combined method is high in sensitivity and selectivity; liquid-liquid extraction can be adopted to remove excess labeling reagent DMPA effectively, Strata X solid phase extraction columns can be used for removing excess activator EDC, quantitative determination of extremely-low-abundance methylation modified cytidylate in living bodies can be realized, LC separation sensitivity and ESI-MS detection sensitivity of nucleotides can be improved obviously, and signal responsiveness is increased by 1 to 2 magnitude orders.

The invention discloses a herpes zoster vaccine composition as well as a preparation method and application thereof, and belongs to the field of vaccines. The vaccine composition comprises varicella-zoster glycoprotein E, a polylactic acid-polyglycolic acid copolymer (PLGA), a double-stranded polycytosine nucleotide fragment (Poly I: C) and a GC-rich single-stranded oligodeoxynucleotide fragment (CpG ODN), the particles with the diameter of 20-400 nanometers are prepared through microfluidic equipment or a double-emulsifying-medium evaporation method. The vaccine composition can specifically enhance humoral immune response and cellular immune response to varicella-zoster glycoprotein E, and can be used as a herpes zoster vaccine; and the components in the vaccine composition are cheap andeasily available, so that the vaccine cost is effectively reduced, and the vaccine yield is increased.

The invention discloses a kit for detecting congenital aniridia pathogenic gene mutation and application of the kit. Detected pathogenic gene mutation is mutation of 140th-position adenine nucleotideof a second exon of a PAX6 gene into cytidylate, namely, NM_000280.3: c.140 A>C. The kit for detecting the congenital aniridia pathogenic gene mutation comprises a 10 * PCR buffer solution, 25 mM dNTPs, DNA polymerase, a PCR amplification primer pair and ddH2O, wherein the PCR amplification primer pair consists of a primer 1 (as shown in SEQ ID NO: 1) and a primer 2 (as shown in SEQ ID NO: 2). Thekit provides a novel congenital aniridia detection means, can provide genetic diagnosis and genetic counseling for clinically conformed congenital aniridia patients, and can provide genetic diagnosisfor eye abnormal patients with undetermined phenotypes to guide clinic treatment.

The invention discloses a feed for treating a fatty liver disease of turtles and a preparation method thereof, and belongs to the technical field of turtle feed processing. The feed is prepared from, by weight, 30-50 parts of silkworm pupa meal, 50-70 parts of manihot utilissima powder, 8-16 parts of table salt, 6-14 parts of dimethyl-beta-propiothetin, 5-9 parts of betaine, 4-8 parts of glycine, 4-8 parts of cytidylic acid, 0.2-0.6 part of Tomatin polypeptide, 1-3 parts of citronellal and 15-42 parts of traditional Chinese medicine preparation. The feed is prepared through the steps of drying, smashing, screening, liquid medicine extracting, concentrating, mixing, pelletizing, sterilizing and the like. According to the feed for treating the fatty liver disease of the turtles, the effects of multiple traditional Chinese medicines are synthesized, the effects of resisting bacteria, diminishing inflammation, enhancing the body immune function and the like can be achieved, the cure rate of the fatty liver disease of the turtles can be effectively increased, and the death rate of the fatty liver disease of the turtles can be effectively decreased; the feed is reasonable in preparation, rich in nutrition and capable of increasing the feed intake of the turtles suffering from the fatty liver disease and increasing the growth speed.

The present disclosure provides modified microRNA nucleic acid compositions that have one or more cytosine and / or uracil bases replaced with gemcitabine or a 5-halouracil, respectively. More specifically, the present disclosure reveals that the replacement of cytosine nucleotides within a microRNA nucleotide sequence with a gemcitabine molecule increases the ability of the microRNA to inhibit cancer progression and tumorigenesis. In addition, the present disclosure reveals that the replacement of cytosine nucleotides within a microRNA nucleotide sequence with a gemcitabine molecule and replacement of uracil bases with 5-halouracil increases the ability of the microRNA to inhibit cancer development. As such, the present disclosure provides various modified nucleic acid (e.g., microRNA) compositions having gemcitabine molecules incorporated in their nucleic acid sequences and methods for using the same. The present disclosure further provides pharmaceutical compositions comprising the modified nucleic acid compositions, and methods for treating cancers using the same.

The invention belongs to the field of biotechnology, and in particular relates to an engineering bacterium containing nCas3 single-stranded endonuclease, a preparation method and an application. This application finds the functional structure division of its Cas3 protein by comparing the differences in the coding gene sequences of endogenous CRISPR-related proteins between the ZM4 strain and other strains that also carry the endogenous type I CRISPR-Cas system. And by exploring the working principle of the I‑F CRISPR‑Cas system, an amino acid in the helicase domain of the Cas3 protein in the ZM4 strain was designed and replaced. The adenine nucleotide number 676,661 in the ZMO0681 gene encoding the Cas3 protein on the original ZM4 strain was replaced with a cytosine nucleotide in situ, so that the Cas3 protein lost the helicase activity and transformed into a single-stranded DNA endonuclease The enzymatically active nCas3 protein was obtained to obtain the modified strain DRM2.

The invention relates to the technical field of preparation of polypropylene anticorrosive materials, and discloses a polypropylene anti-corrosion material based on a biomass cytosine flame retardant.The polypropylene anti-corrosion material comprises the following raw materials in parts by weight: a cytosine flame retardant, polypropylene, an antioxidant and a flexibilizer, wherein the flexibilizer is a mixture of an ethylene-vinyl acetate copolymer, thermoplastic elastomer and powdered rubber. According to the polypropylene anti-corrosion material based on the biomass cytosine flame retardant, a cytosine derivative, cytidylic acid and tris(epoxypropyl)isocyanurate are subjected to a condensation reaction to generate tri(epoxypropyl)isocyanocytidylic acid, and aluminum hydroxide is addedto react with phosphate groups to generate a salt, so that the biomass cytosine aluminum salt with a flame-retardant effect is achieved. The flame retardant improves integrity of the polypropylene material forming a form of expanded carbon, then carbon on the surface layer of carbon layer is compact, and the inner layer is porous, so that flame-retardant efficiency of the polypropylene material is effectively improved, the adding amount is small, and the flame-retardant effect is good.

The invention discloses a preparation method of an electrochemical biosensor for simultaneously detecting Exo I and TdT. Sulfhydryl DNA and a gold electrode surface form an Au-S covalent bond spontaneously by a mercapto part and the sulfhydryl DNA is fixed on the gold electrode surface. Polymerization of the 3'-OH tail of the TdT catalytic sulfydryl DNA and dCTP is carried out and a generated rich-C DNA long chain can used for forming AgNCs; and the TdT activity is detected by a dissolved voltammetric signal of Ag. Before TdT polymerization, Exo I is introduced and the sulfhydryl DNA can be hydrolyzed, so that the formation of AgNCs is affected finally; and thus the ExoI activity is detected. The electrochemical signal intensity can be affected by changing the ExoI or TdT concentration; and on the basis of the principle, a high-efficiency electrochemical biosensor for simultaneously detecting Exo I and TdT is prepared.

The invention belongs to the technical field of biology, and particularly relates to an engineering bacterium containing nCas3 single-stranded endonuclease as well as a preparation method and application of the engineering bacterium. The functional structure division of the Cas3 protein of the ZM4 strain is found by comparing the difference of the coding gene sequences of the ZM4 strain, other strains which also carry an endogenous I-type CRISPR-Cas system and endogenous CRISPR-related proteins. An amino acid in a Cas3 protein helicase structural domain in the ZM4 strain is designed and replaced by exploring the working principle of the I-F type CRISPR-Cas system. According to the method, adenine nucleotides 676 and 661 in ZMO0681 genes for coding Cas3 protein on an original ZM4 strain are replaced with cytosine nucleotides in situ, so that the Cas3 protein loses helicase activity and is converted into nCas3 protein only carrying single-stranded DNA endonuclease activity, and therefore, a modified strain DRM2 is obtained.

The invention discloses a chemical labeling and LC-MS combined method, and applications thereof in nucleotide analysis. According to the chemical labeling and LC-MS combined method, N,N-dimethyl-p-phenylenediamine is used for chemical labeling of 10 monophosphate nucleotides containing two methylation modified nucleotides, so that the retention behavior of the modified nucleotides in reversed phase liquid chromatography is improved obviously, and detection sensitivity of the modified nucleotides in mass spectrometric detection is increased. The chemical labeling and LC-MS combined method is high in sensitivity and selectivity; liquid-liquid extraction can be adopted to remove excess labeling reagent DMPA effectively, Strata X solid phase extraction columns can be used for removing excess activator EDC, quantitative determination of extremely-low-abundance methylation modified cytidylate in living bodies can be realized, LC separation sensitivity and ESI-MS detection sensitivity of nucleotides can be improved obviously, and signal responsiveness is increased by 1 to 2 magnitude orders.

The invention discloses a quality control method for ribonucleic acid II for injection. The method comprises the following steps that the nucleic acid enzyme hydrolysis solution of a substance to be measured is subjected to high-performance liquid chromatogram analysis, an adopted chromatographic column is an Agilent ZORBAX SB-AQC18 chromatographic column, and a flowing phase is a mixture of a formic acid solution and an acetonitrile solution; after the analysis, if the substance to be measured is determined to contain five substances as follows: cytidylate, uridine monophosphate, guanine nucleotide, guanosine and adenosine, the substance to be measured is the ribonucleic acid II for injection or is the ribonucleic acid II for injection as a candidate; and if not, the substance to be measured is not the ribonucleic acid II for injection or is not the ribonucleic acid II for injection as the candidate. A high-performance liquid chromatographic technique is utilized, and the strong-specificity quality control method for the ribonucleic acid II for injection is established. The method has important meanings on increasing the technological content of the medicine, increasing the safety and effectiveness, reducing the cost, enlarging the production scale, increasing the market occupancy, and going forward to the international market.

Methods and kits are provided for testing the functional effect of methylating different cytosine residues, for testing patterns of DNA methylation on gene expression, and for site-specific methylation, as well as methylated DNA constructs. Methods are provided that include steps of denaturing a circular double-stranded DNA construct; hybridizing primers to separate strands of the denatured circular DNA construct; contacting the hybridized primers with a DNA polymerase, deoxynucleoside triphosphates, and copies of the primers; contacting nicked copies with a DNA ligase so as to form a copy of the circular DNA construct; contacting the copy of the circular DNA construct with a methyltransferase enzyme; transfecting a cell with C-5 methylated circular DNA construct; and quantifying expression of a gene of interest, thereby determining the effect of C-5 methylating cytosine nucleotide residues of DNA on expression of the gene of interest.

The present invention relates to an isolated polynucleotide encoding a Na+ / K+ ATPase polypeptide useful in methods to identify therapeutic agents useful for treating cardiovascular diseases, the polynucleotide is selected from the group consisting of:SEQ ID 4 and SEQ ID 5 (baySNP-1765) with allelic variation G in position 240 contained in a functional surrounding like full length cDNA for Na+ / K+ ATPase and with or without the Na+ / K+ ATPase promotor sequence; and SEQ ID 4 and SEQ ID 5 (baySNP-1765) with allelic variation A in position 240 contained in a functional surrounding like full length cDNA for Na+ / K+ ATPase and with or without the Na+ / K+ ATPase promotor sequence.The invention also provides diagnostic methods and kits including antibodies determining whether a human subject is at risk for a cardiovascular disease. The invention provides further polymorphic sequences and other genes.

The invention relates to a compound nucleotide immunoenhancer for paralichthys olivaceus, which is composed of the following substances in percentage by weight: 55-65% of cytidylate, 5-15% of uridine monophosphate, 5-15% of thymidine phosphorylase, 1-5% of hypoxanthine nucleotide and 10-20% of bioactive peptide. The compound nucleotide immunoenhancer disclosed by the invention is capable of obviously improving the ingestion and the growth of paralichthys olivaceus, and remarkably improving the cellular immunity, the humoral immunity, the immune gene expression quantity, the disease resistance and the like of paralichthys olivaceus. Moreover, the compound nucleotide immunoenhancer can also be directly added in a feed for paralichthys olivaceus, and orally fed; and the compound nucleotide immunoenhancer has the advantages of being free from toxic and side residues, free from drug resistance, and pollution-free to environment.