121 results about "DNA unwinding enzyme" patented technology

This invention discloses immunostimulatory combinations of Tumor Necrosis Factor Receptor Superfamily (TN-FRSF) agonists, Toll-Like Receptor (TLR) agonists, “domain present in NAIP, CIITA, HET-E, TP-I (NACHT)-Leucine Rich Repeat (LRR)” or “NLR” agonists, RIG-I-Like Helicase or “RLH” agonists, purinergic receptor agonists and cytokine/chemokine receptor agonists, together with delivery methods. The combinations, when used alone at the site of pathology, provide immunostimulation that induces host humoral and cellular immunologic responses to eliminate pathogens or neoplasms. Alternatively, when the combinations are used with a defined antigens, these combinations can induce focused humoral and cellular immunologic responses useful as prophylactic and/or ameliorative therapeutic modalities for infections and the treatment of neoplastic disorders.

A helicase-mediated amplification method for circular DNA templates and target DNA sequences within the templates is provided. The method combines a DNA polymerase and a helicase preparation to amplify a target sequence as well as the entire circular DNA template.

Provided are compositions that include an at least partially single-stranded nucleic acid, at least one first molecular beacon, and an enzyme comprising a helicase activity, which enzyme is capable of removing the first molecular beacon from the single-stranded nucleic acid wherein the first molecular beacon is hybridized to a first complementary subsequence of the nucleic acid. Also provided are methods of determining the sequence of a template nucleic acid that include removing molecular beacons that are hybridized to the template from the template in a sequential manner using an enzyme that exhibits a helicase activity, detecting a sequence of fluorescent signals that is produced by the removal of a molecular beacons, and converting the sequence of fluorescent signals into nucleotide sequence information. Sequencing systems in which compositions and methods of the invention can be used are also provided.

The invention relates to a new method of characterising a target polynucleotide. The method uses a pore and a RecD helicase. The helicase controls the movement of the target polynucleotide through the pore.

The invention relates to the X-ray crystal structure of the hepatitis C virus helicase domain. More specifically, the invention relates to crystallized complexes of HCV helicase and an oligonucleotide, to crystallizable compositions of HCV helicase and an oligonucleotide and to methods of crystallizing an HCV helicase-oligonucleotide complex. The invention further relates to a computer programmed with the structure coordinates of the HCV helicase oligonucleotide binding pocket or the HCV helicase nucleotide triphosphate pocket wherein said computer is capable of displaying a three-dimensional representation of that binding pocket.

The invention discloses a method for extraction and preservation of helicase. According to the method, a snail is selected and treated, and the digestive juice of the snail is extracted, filtered and then centrifugalized for refinement, and then subjected to a vacuum freeze-drying treatment so as to be packed and stored, with 9 kinds of enzyme valuable in application effectively extracted from the snail digestive gland. Adoption of the method provided in the invention can realize the volume production of helicase which can be preserved for 10 years at a normal temperature, while helicase products still maintain good activity. Therefore, the method has good economic benefit.

The invention relates to the technical field of microbial enhanced oil recovery, and in particular relates to an extraction method of oil reservoir microbial genome DNA. The method comprises the following steps: removing interference of crude oil in samples with extraction by means of organic solvent extraction; releasing microbes on an oil-water interface; filtering water phase by using a hollow fiber membrane to enrich microbes; removing organic macromolecular components such as proteins by phenol and chloroform in cooperation with lysis cells such as lysozyme, helicase, protease and lauryl sodium sulfate according to a physical wall-breaking method under a high-salinity condition; subsequently, performing nucleic acid extraction and precipitation to obtain the oil reservoir microbial genome DNA. The method is quick and simple, and the whole extraction process can be completed in 10 hours; the extraction efficiency is high, and the purity of the obtained genome is high.

The invention discloses a preparation method of recombinant nuclear polyhedrosis virus (NPV) which infects ectropis oblique. The preparation method comprises steps that: (1) recombinant vectors pFastBacDual-polh-helicase are prepared; (2) the recombinant vectors pFastBacDual-polh-helicase are transformed into E.coli DH10 BmBacmid competent cells; the cells are cultivated on a cultivating plate; positive plaques are obtained through blue-white selection; and recombinant BmBacmid-polh-helicase DNA is extracted from the positive plaques; (3) the ecombinant BmBacmid-polh-helicase DNA is transfected to domestic silkworm cultivated cells; cultivated cell supernatant is collected; and recombinant virus BmEoNPV is obtained; (4) the recombinant virus BmEoNPV is used for injection; and recombinant virus polyhedron is obtained. The recombinant nuclear polyhedrosis virus is sprayed on tea tree leaves, such that the ectropis oblique can be prevented from harming the tea trees. The preparation method also has advantages of low cost and controllable productions.

The invention provides an application of helicase in directionally hydrolyzing rutin for biosynthesizing Quereetin. The method provided by the invention comprises the following steps: adding a crude helicase preparation in rutin substrate solution with the concentration of 0.005-0.100g/L, enabling the helicase concentration in the solution to be 0.1-50g/L and the pH value to be 3-9, performing enzymatic hydrolysis reaction under the conditions that the reaction temperature is 10-70 DEG C and the reaction time is 0.1-36h, and acquiring the hydrolyzed product Quereetin after reaction. The invention adopts the crude helicase preparation or immobilized helicase preparation to directionally catalyze and hydrolyze rutin for biosynthesizing Quereetin, thus having the advantages of wide sources of catalyst, easy preparation and low cost, and the helicase preparation has high stability, can be stored easily, has high catalytic efficiency and high specificity. Therefore, by utilizing the helicase, the production cost of the Quereetin can be greatly reduced, the enzymatic conversion rate is high and the product is specific.

The invention discloses a helicase-dependent isothermal DNA amplification-based method for detecting microRNA. The method comprises the following steps: (1) extracting total RNA in a sample; (2) adding a single-strand DNA probe and excessive exonuclease I into the extracted total RNA, incubating in a reacting buffer solution to realize specific combination of target microRNA and the single-strand DNA probe, and removing residual single-strand DNA probe by utilizing the excessive exonuclease I; and (3) adding upstream primer, downstream primer, single-strand binding protein and helicase to the reaction system, amplifying the target microRNA, and detecting expression of the target microRNA through a fluorescent signal. According to the method, the background is reduced by digesting the exonuclease I, and the signal is amplified by means of exonuclease assisted isothermal amplification (HDA) reaction, so that the microRNA can be quickly and sensitively detected.

Inhibitors of DNA damage polymerases, endonucleases, and helicases are provided. In particular, compounds comprising Formula (I) are described.

The invention relates to a salmonella nucleic acid rapid detection kit, a salmonella nucleic acid rapid detection test strip and a salmonella nucleic acid rapid detection method, and relates to design of salmonella specific primers, optimization of helicase isothermal amplification conditions and establishment of a nucleic acid film chromatography detection test strip, and visual observation of a nucleic acid detection result is achieved. Just one thermostat is adopted in a whole reaction process, and from pretreatment to detection completion on a single sample, the detection result can be obtained just by 1h; the method, on detecting salmonella, is 4.5*10<1>CFU/mL in sensitivity, and the method can avoid a cross reaction with other common food-borne pathogenic bacteria; and the detection test strip is simple and convenient to operate, high in sensitivity, strong in specificity and high in efficiency, and the detection strip is especially suitable for on-site detection and clinical diagnosis in grassroots units.

The invention relates to a preparation method of ganoderma polysaccharide, particularly relates to a preparation method of breeding high-yield extracellular polysaccharide strains by using lithium chloride to mutagenize red ganoderma bioplast, belongs to genetic technology field. The invention uses reg ganoderma as strains, activation by potato dextrose agar medium, starch liquid shaking cultivation, using lywallzyme, helicase and cellulase to hydrolyze red ganoderma mycelium, shaking at constant temperature to prepare red ganoderma bioplast. After mutagenesis of lithium chloride, content of extracellular polysaccharide is improved 8 to 10 times than original strains. After generation of the mutagenized strains, content of extracellular polysaccharide is improved 6 to 8 times than original strains, and its genetic character is stable, the engineering strain can be used in industrial production of ganoderma polysaccharide.

The invention discloses a reagent kit for quickly detecting nucleic acid of canine parvovirus (CPV). The reagent kit comprises lysate, diluent, lyophozyme tubes with primers, detection liquid and test strips. The reagent kit has the advantages that normal-temperature nucleic acid amplification technologies are adopted, different types of enzymatic reaction can be carried out by the aid of DNA (deoxyribonucleic acid) helicase, single-stranded DNA binding proteins, DNA polymerase and the like, and accordingly to-be-detected DNA can be amplified by several millions of times under the condition of the constant temperatures of 37-45 DEG C within 10-30 minutes; the reagent kit is matched with immune colloidal gold chromatographic detection technologies, and accordingly the to-be-detected DNA can be quickly verified; the reagent kit is easy to operate, short in time and quite suitable for quickly diagnosing infectious diseases of pets and has low requirements on instruments.

The invention discloses an isothermal amplification nucleic acid detection method based on helicase and nicking enzyme. The isothermal amplification nucleic acid detection method comprises the following steps that S1, a specific amplification primer and a Taqman probe with nicking enzyme recognition sites are designed according to a target nucleotide sequence; S2, a sample nucleic acid is subjected to enzyme treatment to obtain a single-chain nucleic acid template; S3, the Taqman probe obtained in the step S1 and the single-chain nucleic acid template obtained in the step S2 are specifically bonded to obtain a specific bonding product, the specific bonding product is digested by using the nicking enzyme, and real-time detection of fluorescence signals is achieved. The helicase and nickingenzyme are used jointly, the Taqman probe with nicking enzyme recognition sites is designed, a large number of nucleic acid products do not need to be amplified, only an appropriate amount of single-chain templates is needed, and then detection can be performed by using the nicking enzyme to constantly cut the probe bound with recognition sites, the shortcoming of lab pollution likely caused by isothermal amplification can be avoided, and the isothermal amplification nucleic acid detection method is simple and quick to operate and suitable for on-site quick detection.