538 results about "Ribonucleotide synthesis" patented technology

The present invention relates generally to methods, compositions and kits for synthesizing sense RNA molecules from one or more RNA molecules of interest in a sample. In exemplary embodiments, the methods use a terminal tagging oligoribonucleotide (rTTO) to join a DNA sequence tag to the 3′-termini of first-strand cDNA molecules. The use of an rTTO comprising ribonucleotides results in decreased oligonucleotide-derived background synthesis of RNA in the absence of sample RNA and, surprisingly and unexpectedly, also results in significantly increased yields of sense RNA molecules that exhibit sequences that are substantially identical to those of the RNA molecules of interest in the sample. The sense RNA molecules also have an RNA sequence tag on their 5′-termini that is useful for fixing the lengths of sense RNA molecules that are synthesized in a second or subsequent round.

A method of detecting the presence of an ion includes contacting a nucleic acid enzyme with a sample suspected of containing the ion, where the enzyme contains a ribonucleotide and is dependent on the ion to produce a product from a substrate, and measuring an amount of the product produced. The ion is Pb<2+>, and is in the presence of other ions.

The present invention comprises methods and compositions for splint-assisted enzymatic synthesis of polyribonucleotides using an RNA polymerizing enzyme. The invention provides ligating ribonucleotides comprising ligating a donor RNA molecule to an acceptor RNA molecule in the presence of RNA ligase and a splint, wherein the donor RNA molecule is comprised of at least one nucleotide and a ligation linker moiety, the acceptor RNA molecule is comprised of at least one nucleotide and a ligation linker moiety and the splint is comprised of a polyribonucleotide. The invention also provides splints for use in splint-assisted enzymatic synthesis using an RNA polymerizing enzyme.

Methods are provided for amplification of a nucleic acid sequence. The method use RNA / DNA chimeric oligonucleotides as primers. The primers have RNA residues scattered along their length and no two ribonucleotides in the prime are adjacent to one another. The methods are useful for reducing non-specific amplification products, such as primer dimers. The invention also provides kits comprising RNA / DNA chimeric oligonucleotide primers for practicing the amplification methods.

The present invention provides methods for synthesis and therapeutic use of DNA and RNA oligonucleotides and analogs. RNA oligonucleotides are synthesized using a small, circular DNA template which lacks an RNA polymerase promoter sequence. The RNA synthesis is performed by combining a circular single-stranded oligonucleotide template with an effective RNA polymerase and at least two types of ribonucleotide triphosphate to form an RNA oligonucleotide multimer comprising multiple copies of the desired RNA oligonucleotide sequence. Preferably, the RNA oligonucleotide multimer is cleaved to produce RNA oligonucleotides having well-defined ends. Preferred RNA oligonucleotide multimers contain ribozymes capable of both cis (autolytic) and trans cleavage.

The invention relates to a double-stranded RNA (dsRNA) molecule comprising between about 15 base pairs and about 40 base pairs, wherein at least one ribonucleotide of the dsRNA is a 5′-methyl-pyrimidine, and a method of using such modified dsRNA molecule to increase stability of RNA when in contact with a biological sample.

The invention relates to a compound mushroom flavoring and a preparation method thereof. The main raw materials of the compound mushroom flavoring comprise meadow mushroom powder, straw mushroom powder, sodium glutamate, salt, sugar, maltodextrin, yeast extract, disodium 5'-ribonucleotide and yolk powder. The preparation method of the compound mushroom flavoring comprises the following steps: rinsing, boiling, pulverizing, pulping, homogenizing, concentrating and spray-drying fresh mushrooms, and then, obtaining mushroom powder; adding other raw materials; and carrying out the processes of mixing, granulating, drying and the like to obtain the compound mushroom flavoring. The compound mushroom flavoring has the flavor of the meadow mushroom and the straw mushroom simultaneously by organically mixing the sodium glutamate, the yeast extract, the disodium 5'-ribonucleotide and the mushroom powder, has strong functions of increasing freshness and increasing fragrance and has soft flavor. The cell walls of mushrooms can be broken by the homogenizing action to better release the substances in the cells of the mushrooms, thereby further improving the nutrient value of the product. The sugar is added in the concentration process, thereby further improving and increasing the flavor of the materials. The compound mushroom flavoring is granular and is convenient in use.

Methods of amplifying a target nucleic acid whereby the target nucleic acid is amplified in the presence of a deoxyribonucleotide triphosphate, a DNA polymerase having strand displacement activity, at least one chimeric oligonucleotide primer, at least one upstream block oligonucleotide and a RNase H; wherein the chimeric oligonucleotide primer contains a ribonucleotide positioned at the 3′-terminus; wherein the upstream block oligonucleotide is capable of annealing to a region 3′ to a portion in the nucleic acid as the template to which the chimeric oligonucleotide primer anneals; and compositions and kits thereof.

The invention relates to a double-stranded RNA (dsRNA) molecule comprising between about 15 base pairs and about 40 base pairs, at least one 5′-methyl-pyrimidine and at least one 2′—O-methyl ribonucleotide, and a methods of using such modified dsRNA molecule to increase stability of a siRNA molecule when in contact with a biological sample, to reduce off-target effects and to reduce interferon responsiveness (IFN) of the siRNA molecule.

A method for cloning and expressing a beta-glucosaccharase gene and application in preparing gentian oligose belongs to the field of enzyme gene engineering and enzyme engineering. In the invention, cDNA of beta-glucosaccharase gene (bgl)SEQ ID NO: 1 synthesized by the reverse transcription of the gross RNA of Aspergillus niger WX-07 uses plasmid pPIC9K as an expression vector and P.pastoris as an expression host to realize the solubility expression of the bgl gene outside a cell; the full length of the cDNA of the bgl is 2523 ribonucleotide and 841 amino acids are coded; the constructed bgl / pPIC9K transforms P.pastoris KM71 so that P.pastoris KM71 can express BGL enzyme. The BGL enzyme has the activity of transglycosylation and can generate dextrose into the gentian oligose by transglycosylation. The technique of preparing the gentian oligose by enzyme process is optimized; and resin cation is used to carry out separation and refining to achieve a better effect. The prepared gentian oligose product, as functional food ingredient, has the physiological functions of low energy, low decayed tooth and stimulating the gastrointestinal mucosa. The invention provides a new approach with commercial values for preparing the gentian oligose.

The invention discloses cathelicidin-BF and a gene and application thereof, which belong to the field of biomedicine. The cathelicidin-BF is straight chain polypeptide and contains thirty amino acid residues, the molecular weight is 3,637.54Da, and the isoelectric point is 11.79. The complete sequence of the cathelicidin-BF is lysine-phenyl alanine-phenyl alanine-arginine-lysine-leucine- lysine-lysine-serine-valine-lysine-lysine-arginine-lactamine-lysine-glutamic acid-phenyl alanine- phenyl alanine-lysine-lysine-proline-arginine-valine-isoleucine-glycin-valine-serine-isoleucine- praline-phenyl alanine. The gene for encoding the cathelicidin-BF consists of 750 ribonucleotides, wherein 484th to 573rd ribonucleotides are used for encoding a mature peptide part. The cathelicidin-BF has small molecular weight, strong sterilization effect, and quick action time, and has quite strong killing function to a plurality of kinds of clinical drug-fast bacteria. In addition, the cathelicidin-BF also has the advantages of broad-spectrum antibiotics, salt independence and so on.

The present invention provide a process for sequence-specific nucleic acid amplification capable of improving the detection sensitivity and increasing the signal. In particular, the present invention provides a reagent for nucleic acid amplification containing at least one member selected from the group consisting of EDTA, NTA, UDA, CyDTA, DTPA, GEDTA, TTHA and their salts, specifically a reagent for nucleic acid amplification comprising, in addition to the at least one compound, a forward primer having a DNA sequence homologous to a sequence of a target RNA; a reverse primer having a DNA sequence complementary to a sequence of the target RNA and having a promoter for RNA polymerase attached to its 5' end; ribonucleotides; deoxyribonucleotides; a reverse transcriptase or RNA-directed DNA polymerase; a RNase H; a DNA polymerase or a reverse transcriptase having DNA-directed DNA polymerase activity; and a RNA polymerase. The present invention also provides a process for nucleic acid amplification characterized by carrying out a nucleic acid amplification reaction in the presence of at least one member selected from the group consisting of EDTA, NTA, UDA, CyDTA, DTPA, GEDTA, TTHA and their salts.

Methods for identifying ribonucleotide sequences, in vitro, using the ribosome-mediated translation, are provided.

The methods of the present invention allow for the measurement of ribonucleotide reductase (RR) activity, an important enzyme in the de novo DNA synthesis pathway. Ribonucleotide reductase converts all four ribonucleotides to their deoxy form and is a rate-controlling step in this pathway. Biosynthetic pathways of deoxyribonucleotides (dN) have received considerable attention in the context of anti-proliferative chemotherapy. Inhibitors of various steps in dN biosynthesis, including inhibitors of RR are among the most useful chemotherapeutic agents in cancer, viral infections, and other therapeutic uses. DNA synthesis from the dN salvage pathway is also an important component to DNA replication. The relative contributions from RR vs. salvage pathways are critical to the actions and effectiveness of chemotherapeutic agents that act on nucleoside metabolic pathways. Until now, however, it has not been possible to study these metabolic processes in vivo. Disclosed within are methods of measuring RR activity in vivo and in vitro which find use, among other things, in drug discovery, development, and approval.

The present invention relates to chimeric molecules comprising an oligonucleotide complementary to a region of the ribonucleotide component of telomerase attached to an activator of RNase L ("activator-antisense complex") which specifically cleaves the ribonucleotide portion of a telomerase enzyme. The present invention relates to methods of inhibiting telomerase enzymatic activity with activator-antisense complexes targeted to the RNA component of telomerase. The present invention further relates to methods of treating malignant neoplastic disease, wherein the malignant cells contain a telomerase activity that is necessary for the growth of the malignant cells.

The invention discloses a bone soup seasoning, which is characterized by comprising the following raw materials in parts by weight: 45-50 parts of fresh bones, 6-10 parts of pigskins, a right amount of white sugar, a right amount of salt, a right amount of powdered monosodium glutamate, 2-4 parts of disodium 5'-ribonucleotide, 2-3 parts of mashed garlic, 4-5 parts of pitaya flowers, 3-4 parts of peach leaves, 2-3 parts of ginger, 10-15 parts of starches, 1-2 parts of Rhodiola rosea, 3-5 parts of agaricus blazei murill, 1-2 parts of bengal waterdropwort herbs, 2-3 parts of hericium, 3-5 parts of rhizoma ligustici wallichii and 0.08-0.12 part of xanthan gum. Through adding traditional Chinese medicinal materials such as pitaya flowers, peach leaves and rhodiola rosea, the seasoning disclosed by the invention not only has the characteristics that the seasoning is fresh, fragrant, delicious and rich in tastes, but also enhances the health care value, and further enhances the health-care benefits of the product.

The invention discloses hotpot condiment and a preparation method. The method comprises the following steps: heating A grade rapeseed oil to 185-195 DEG C, adding dry red pepper powder, and stir-frying for 4 min at 175-185 DEG C; adding Pixian bean paste and fermented soya beans, and stir-frying for 3 min at 95-105 DEG C; adding ginger granules and garlic granules, and stir-frying for 4 min at 90-100 DEG C; adding pickled ginger and ciba pepper, and stir-frying for 2 min at 75-85 DEG C; adding compound spices and white granulated sugar, and stir-frying for 1 min at 80-90 DEG C; adding green pepper powder and red pepper powder, and stir-frying for 1.5 min at 75-85 DEG C; adding edible salt and monosodium glutamate, and stir-frying for 1 min at 80-90 DEG C; adding white spirit, millet wine, disodium 5'-ribonucleotide, yeast extract, edible essence, scallop powder, capsanthin and fermented glutinous rice, stir-frying for 1 min at 75-85 DEG C, and uniformly stirring the components. The hotpot is red in oil color, clear in soup color, spicy, fragrant, free of greasy feeling after eating for a long time and is led up to meaningful afterthoughts, thereby being well praised by consumers.

The invention provides a recombinant microorganism for producing cytidine and a method for producing the cytidine from the recombinant microorganism. A degradation and use gene of the cytidine is knocked off, and the gene encodes cytidine ammonialyase, ribonucleotide hydrolase, cytidine / uridine kinase and nucleoside transporter. Meanwhile key enzymes in biological synthesis process of the cytidine are over-expressed, including degrading cytidine triphosphoric acid to cytidine triphosphoric acid pyrophosphorylase of the cytidine monohosphoric acid, and catalyzing the cytidine monohosphoric acid to cytidine monohosphoric acid phosphorylase of the cytidine. In addition, pyrimidine nucleoside process is subjected to genetic engineering reform, and feedback inhibition of the synthesis process is relieved. By using a biological fermentation method, the cytidine yield greater than 20g / L can be achieved for a recombinant strain in a fermentation tank of 5L, industrial on-scale production can be achieved, and meanwhile the recombinant microorganism is low in cytidine production cost, small in pollution, green and environmental-friendly and relatively high in popularization and application value.

A protective group represented by the following general formula (I) (the oxygen atom attached with * represents oxygen atom of 2′-hydroxyl group of a ribonucleoside, a ribonucleotide or a derivative thereof; R1 and R2 both represent hydrogen atom, or represent a halogen atom, a C1-6 alkyl group, or a C1-6 halo-substituted alkyl group; R3 and R4 represent hydrogen atom, a halogen atom, a C1-6 alkyl group, or a C1-6 halo-substituted alkyl group; and R5 and R6 represent a halogen atom, a C1-6 halo-substituted alkyl group, cyano group, nitro group, or the like), which is stable under the reaction conditions of the nucleic acid synthetic cycles and has little steric hindrance, and can be removed under mild conditions using fluoride ions as a base.

Point-of-care binding assays include at least one target nucleic acid binding in a multiplex structure with at least one sequence in a partner nucleic acid associated with a label, due to complementary base pairings between at least one sequence in the target nucleic acid and at least one sequence in the partner nucleic acid. The assays overcome the inherent deficiencies of antibody-protein antigen assays. In a preferred embodiment, color tagged nucleic acid sequences are used to bind a complementary target nucleic acid. The tagged nucleic acid sequences are preferably made from deoxyribonucleotides, ribonucleotides, or peptide nucleotides.

The invention provides a chicken flavor seasoning and its preparation method. The chicken flavor seasoning is prepared by the following raw materials of: by weight, 5-15% of a dry chicken powder, 2-5% of refined chicken oil, 3-10% of a chicken bone extract product, 3-10% of a chicken extract product, 0.3-0.8% of disodium 5'-ribonucleotide, 20-40% of salt, 5-10% of a granulated sugar powder, 3-10%of glucose, 0.1-0.3% of a turmeric powder, 0.5-2% of ginger juice, 0.5-2% of shallot juice, 1-2% of an onion powder, 0.1-1% of a white pepper powder, 3-10% of glutamic acid, 2-10% of maltodextrin, and 0.5-1% of an anticaking agent. The above raw materials are mixed, dried and crushed to obtain the chicken flavor seasoning which has a strong and natural taste and is a strong cooking seasoning. Thechicken flavor seasoning is simple to prepare; the production period is short; and the finished product has high quality, and is convenient and fast to store and use.

The invention mainly relates to detection of poultry-derived component, which mainly relates to detection of poultry-derived component by using PCR-mtDNA. A specific PCR-mtDNA amplification primers for detecting poultry-derived component is characterized by compring a upstream ribonucleotide with length of 18bp, a downstream ribonucleotide with length of 19bp, and the sequence of the upstream primer PF(18bp) is 5'-AGAACTACGAGCACAAAC-3', and the sequence of the downstream primer PR(19bp) is 5'-GCTATACCTTGACCTGTCT-3'. The detection of poultry-derived component by using PCR-mtDNA has the advantages that: the method is an accurate, rapid and reliable identification method and technology of poultry-derived component, and has the important practical significance of effective resistance on propagation and spread of bird flu and other poultry diseases. The research utilizes aseptic ultrapure water to dilute DNA template of poultry to cause the concentration to decrease to 12ng per Mul, 1.2 ng per Mul, 120 pg per Mul, 12 pg per Mul, 1.2 pg per Mul, 120 fg per Mul, 12 fg per Mul and 1.2 fg per Mul, and carries out PCR amplification. Known from the electrophoresis result of PCR product, the minimum concentration which can be detected is 120 pg per Mul, so the sensitivity of the primer is 120 pg per Mul.

The invention discloses a seasoning processing technique for spicy capsaicin, comprising the following steps of: soaking: soaking textured soybean protein by cold water, wherein the temperature of cold water is 10-15 DEG C and the soaking time is 4-6hours; dehydrating: dehydrating the soaked textured soybean protein till the water content is 45-50%, wherein the dehydration mode is natural drying dehydration or mechanical centrifugal dehydration; frying: slicing the hydrated textured soybean protein to obtain original capsaicin, and frying by a continuous automatic fryer, wherein the frying temperature is controlled at 145-170 DEG C, and the frying time is 70s; blending: adding seasoning in the fried capsaicin and blending. By strictly controlling the technical parameters of the seasoning processing technique, the produced product is even in color and luster, good in taste, more delicious after seasoning of a special formula is added, and is safe and healthy to eat, and only a food additive disodium 5'-ribonucleotide is added.

The invention discloses a seasoning powder with natural strong mushroom flavor and a preparation method thereof. The method comprises the steps of using Agaricus bisporus precooked water or extract, condensing, adding a certain amount of monosaccharide, adjusting the pH to be 6-7, and performing the Maillard reaction at 65-75 DEG C for more than 6 hours to obtain a concentrated solution with the natural strong mushroom flavor, then adding maltodextrin, dissolving the maltodextrin completely, performing spray drying to obtain concentrated mushroom powder, and finally, mixing the concentrated mushroom powder, common salt, sodium glutamate, sucrose, flavor-development disodium ribonucleotide, supercritical CO2 extract of spice, calcium phosphate and starch-maltodextrin mixture, adequately agitating until above materials are mixed uniformly, granulating and drying to prepare the seasoning powder with natural strong mushroom flavor.

The invention discloses crispy black rice and a method. The crispy black rice comprises the following components in weight portion: 25-35 portions of black rice, 55-65 portions of corn meal, 0-10 portions of soybean and 3-5 portions of seasoning, wherein the seasoning comprises the following components in weight portion: 1.2-1.4 portions of hot pepper, 1.1-1.3 portions of salt, 2-3 potions of white sugar, 0.2-0.3 portion of monosodium glutamate, 0.01-0.03 portion of disodium 5'-ribonucleotide, 0.05-0.55 portion of cumin powder, 0.1-0.2 potion of Chinese prickly ash, 0.1-0.2 portion of white pepper, 0.5-0.55 portion of maltodextrin and 0.04-0.08 portion of aniseed. The method comprises the following steps: soaking, cleaning, steaming, cooking and pulverizing the black rice and the soybean, then stirring with the corn meal, molding, cutting into pieces; frying by oil, cooking, and mixing with seasoning to obtain a finished product. The crispy black rice has high nutritive value and unique taste, and is crispy; and the preparation method is simple and convenient to operate.

The invention discloses a method for brewing composite selenium-enriched mushroom nutritious soy sauce. The method takes mushroom leftovers to partly replace soybean cakes for making starters and hydrolyzes the other part of the mushroom leftovers through proteinase; then the hydrolysate solution and the starters are mixed round and are fermented so that the rich protein, amino acid, flavor developing ribonucleotide, mushroom amylase and vitamins can thoroughly enter the soy sauce mash; and active slenium ferment rich in organic selenium is added into the soy sauce mash in the post-maturation process. A small amount of aroma-enhancing substances like ethanol and ester are produced during the fermentation process and the slenium ferment is totally autolyzed in the soy sauce mash. In this way, characteristic high-quality soy sauce can be prepared, which is rich in biologic selenium, amino acid, ribonucleotide, mushroom amylase and microelements and has the unique flavor of mushroom. Meanwhile, the method is good for recovering mushroom leftovers, thus reducing environment pollution and saving a large amount of soybean material.

The invention provides novel dye-labeled ribonucleotide analogs and methods for synthesizing those analogs. The compounds of the invention are especially useful for DNA sequencing by the polymerase chain reaction.