107 results about "Ribonuclease" patented technology

The invention discloses an extraction method and application of high-molecular-weight turbellarian worm genome DNA, and belongs to the field of turbellarian worm genome extraction.The turbellarian worm genome DNA extraction is achieved by improving a lysate formula and removing ethylenediaminetetraacetic acid and increasing Mg < 2 + > on the basis of a conventional reagent in a classic Tris-HCl-phenol-chloroform method to inhibit activity of deoxyribonuclease II in a turbellarian worm body to inhibit degradation of genome DNA, and the high-molecular-weight turbellarian worm genome DNA is obtained. Meanwhile, ribonuclease A is added, RNA in cells can be degraded in the tissue cell digestion process, so that the step of two times of purification is omitted, the loss of DNA yield is reduced, experimental steps are simplified, especially the step of DNA purification is reduced, the problem of degradation in the extraction process of the turbellarian turbellarian worm genome DNA is solved, and the genome DNA yield (gt; 10 micrograms per turbellarian worm), and the turbellarian worm genome ultra-long sequencing or PacBio high-fidelity sequencing requirements are met.

The invention relates to the technical field of medicines, and discloses a stress particle targeted degradation agent as well as a preparation method and application thereof. The compound provided by the invention comprises a ternary complex composed of a stress particle protein, a linking group and an E3 ubiquitin ligase ligand or a ternary complex composed of a stress particle protein, a linking group and a ribonuclease L ligand, the compounds connected with different ligands have good targeting and degradation effects on the stress particles, can be used as a stress particle targeting degradation agent to be applied to preparation of anti-tumor drugs and anti-neurodegenerative disease drugs, and have good application prospects in treatment of stress particle induced tumor drug resistance and neurodegenerative diseases.

The invention provides a respiratory system fungal infection detection system, a kit and a preparation method thereof, and belongs to the technical field of biological detection. The primer probe composition used by the respiratory system fungal infection detection system comprises upstream and downstream primers and a fluorescent probe sequence designed aiming at paracoccus braziliensis, upstream and downstream primers and a fluorescent probe sequence designed aiming at histoplasma capsulatus, and upstream and downstream primers and a fluorescent probe sequence designed aiming at troptosporidium. The kit comprises upstream and downstream primers and fluorescent probe sequences designed for budgettes dermatitis, upstream and downstream primers and fluorescent probe sequences designed for Sirpolyspora sessilifolia, upstream and downstream primers and fluorescent probe sequences designed for talaromyces marneffei, upstream and downstream primers and fluorescent probe sequences designed for internal reference designed for ribonuclease P, upstream and downstream primers and fluorescent probe sequences designed for ribonuclease P, upstream and downstream primers and fluorescent probe sequences designed for ribonuclease P, and upstream and downstream primers and fluorescent probe sequences designed for ribonuclease P, the kit can be used for effectively detecting the fungi of the respiratory system, and ensures high accuracy, specificity and sensitivity.

The invention discloses an application of ribonuclease SiRe0902 with high efficiency and thermal stability in specific recognition of a GG site. The amino acid sequence of the gene is as shown in SEQ ID NO.2, and the nucleotide sequence of the gene is as shown in SEQ ID NO.1. The ribonuclease SiRe0902 has very efficient and specific RNA (Ribonucleic Acid) hydrolysis activity, and does not have non-specific cutting on DNA (Deoxyribose Nucleic Acid) at all. In addition, the enzyme is derived from thermophilic archaea, is detected to have good thermal stability, and has very good RNA cleavage activity under the conditions of 37 DEG C and 65 DEG C. Through detection, the enzyme can specifically recognize a GG sequence for cutting, and also has high cutting activity for the GG sequence in a secondary structural region. Therefore, the ribonuclease SiRe0902 is novel ribonuclease with thermal stability, has extremely high RNA hydrolytic activity under normal-temperature and high-temperature conditions, and meets the requirements in the fields of clinic, production, molecular cloning and the like.

The invention belongs to the technical field of organoid culture, and particularly relates to tissue digestive juice for organoid culture as well as a preparation method and application of the tissue digestive juice. According to the tissue digestive juice, a DMEM / F-12 culture medium which contains L-glutamine and does not contain 4-hydroxyethyl piperazine ethanesulfonic acid is used as a basic culture medium, calcium chloride, collagenase I type, collagenase III type, V type hyaluronidase, deoxyribonuclease I, 4-hydroxyethyl piperazine ethanesulfonic acid and Y-27632 are combined and have a synergistic effect, cell damage is reduced, meanwhile, the tissue digestive juice has the advantages that the tissue digestive juice can be used for improving the tissue digestive juice quality, and the tissue digestive juice has a good application prospect. The digestion time is shortened, the tissue digestion efficiency is improved, and the culture success rate of the organoid is improved.

The present application relates to a deoxyribonuclease mutant and its application in preparing PDRN. The amino acid sequence of the deoxyribonuclease mutant is shown as SEQ ID NO. 1, and the nucleotide sequence of the encoding gene is shown as SEQ ID NO. 2. The present application also provides the application of the deoxyribonuclease mutant and / or the encoding gene of the deoxyribonuclease mutant in preparing PDRN, and the specific application method. The present application is based on the deoxyribonuclease expressed by the DNASE1 gene of bovine pancreas tissue, and the amino acids at positions 69, 132 and 215 are selected for iterative saturation mutation. Compared with the wild-type enzyme, the enzyme activity of the deoxyribonuclease mutant A69H-S132R-Q215C is increased by 2.15 times, reaching 4.1x10 6 U / mL, and the DNA extraction rate of salmon PDRN prepared by using the mutant enzyme prepared by the present application is 15.2%, the DNA content is 99.1%, and the molecular weight is 1.05 million Daltons, which is significantly improved compared with the wild-type enzyme, and the PDRN in this molecular weight range has more significant biological activity.

The present disclosure describes methods, kits, and systems for digesting polyribonucleotides. The methods involve selectively forming an oligonucleotide (e.g., DNA: RNA or RNA: RNA) duplex with a single-stranded target RNA, and then selectively cleaving the target RNA into smaller fragments using a sequence-specific nuclease that acts only on the RNA within the duplex. Additional sequence-specific ribonucleases can be used to provide additional cleavage of the target RNA at a predetermined site. By forming double strands to increase the availability of nuclease enzymes that can be used to cleave the single-stranded target RNA and to selectively control the location at which the target RNA is cleaved, the target RNA can be digested into fragments within a controllable size range that are optimal for polynucleotide analysis, such as by liquid chromatography and mass spectrometry.

The present disclosure is generally directed to anchoring bridge nucleic acid probes in a polymer matrix, which are configured to hybridize to cellular nucleic acid, embedding in a polymer matrix, clearing cellular components including the cellular nucleic acid, contacting the anchored bridge probes with primary probes that hybridize with the bridge probes and imaging. The disclosure describes methods for imaging cellular RNA in an RNase insensitive method.

The invention provides application and a method for repairing growth defects of neuronal dendrites by using ribonuclease RNase H. In particular, ribonuclease RNase H can be used as an R-loop degradation agent for treating and / or preventing neuron dendritic growth defects and related diseases thereof. A Rett syndrome cell model is utilized, and it is found that growth of dendrites can be restored by expressing ribonuclease RNase H of mammals. The method disclosed by the invention provides a very significant means for treating and preventing the Rett syndrome and other neuronal dendritic growth defect related diseases.

The present invention relates to the use of ribonuclease A in preparing a drug for treating sepsis or sepsis complications. The present invention simulates a sepsis model in an animal, injects ribonuclease A into the tail vein for treatment, and ultimately determines the relevant pathways of ribonuclease A in treating sepsis by observing the survival rate of septic mice and combining methods such as tissue pathology and flow cytometry. The test results show that ribonuclease A can effectively treat sepsis. In addition, the test found that ribonuclease A can effectively reduce early neutrophil infiltration into tissues and reduce NETs formation to alleviate the degree of tissue damage. This shows that ribonuclease A can be used for the preparation of drugs for treating or preventing sepsis-related diseases, and has broad application prospects.

This invention relates to the field of pharmaceutical technology, and discloses a stress particle targeted degrader, its preparation method, and its applications. The compounds provided by this invention include ternary complexes composed of stress particle protein, a linker group, and an E3 ubiquitin ligand, or ternary complexes composed of stress particle protein, a linker group, and a ribonuclease L ligand. Compounds with different ligands exhibit good targeting and degradation effects on stress particles and can be used as stress particle targeted degraders in the preparation of antitumor drugs and anti-neurodegenerative disease drugs, showing promising application prospects in treating stress particle-induced tumor drug resistance and neurodegenerative diseases.

The application discloses a single cell whole genome amplification sequencing method. The application comprises the following steps: 1) separating a single cell, adding a single cell lysis solution to sufficiently lyse the single cell and digest proteins combined on the genomic DNA; 2) using a Tn5 transposome with a specific adapter sequence to fragment the DNA, while adding adapters to both ends of the fragments; 3) after end repair, using a ribonuclease to cut modified ribonucleotide residues on the adapter sequence; 4) using the adapter sequence as a primer to linearly amplify the fragmented DNA by using a single primer PCR method; and 5) adding sequencing adapters to both ends of the DNA fragments by using a chain extension and PCR method to obtain a final sequencing library. The method disclosed by the application can amplify single cell whole genome DNA with high coverage, uniformity and fidelity, and simultaneously and accurately detect chromosomal copy number variation CNV and single nucleotide mutation SNV on a single cell genome.

The invention belongs to the technical field of biology, and particularly relates to an efficient extraction method of mineralized tissue RNA applicable across animal categories. The invention provides a solution combination suitable for mineralized tissue pretreatment, which not only is beneficial to promoting the release of cells from a mineralized matrix, but also can inhibit the activity of ribonuclease to a certain extent, and is beneficial to extracting high-quality RNA (Ribonucleic Acid). Furthermore, the invention also provides an efficient extraction method of mineralized tissue RNA suitable for cross-animal categories, the sample is pretreated by using a solution combination, and then RNA extraction, precipitation and other treatments are sequentially performed, so that the RNA extraction efficiency of the exoskeleton sample is remarkably improved, and the method has good universality and is suitable for large-scale popularization and application. The method is suitable for extracting RNA in cross-animal mineralized tissues, has important biological significance, and lays a methodological foundation for subsequent development of transcriptomics research of exoskeleton dynamic development, analysis of an environmental adaptation mechanism and construction of a biomineralization process regulation and control network.

Provided are the use of a ribonuclease RNase H in repairing growth defects of neuron dendrites, and a method therefor. Specifically, the ribonuclease RNase H can be used as an R-loop degradation agent for treating and / or preventing growth defects of neuron dendrites and related diseases thereof. By means of utilizing a Rett syndrome cell model, it is found that expressing the mammalian ribonuclease RNase H can restore the growth of dendrites, thus providing means of great significance for treatment and prevention of the Rett syndrome and other diseases related to growth defects of neuron dendrites.

The invention relates to the field of biotechnological compounds used in the treatment of neurodegenerative diseases. In Alzheimer's disease, oxidative stress, mitochondrial dysfunction, and neuroinflammation are interconnected. These are the mechanisms that nourish; microRNAs can regulate these pathways simultaneously, but in their free state... It is broken down by ribonucleases and cannot cross the cell membrane. The invention solved this problem by sequentially extracting it from mammalian milk. small extracellular vesicles isolated by centrifugation, ultracentrifugation and size-reference chromatography, Synthetic miR-25802 mimic with SEQ ID NO:1 sequence in the presence of 0.15% to 0.25% by weight of saponin. It resolves through active loading. The loading process increases the average particle diameter of the vesicle and It does not change its hydrodynamic diameter; the mimic remains protected from ribonucleases within the vesicle lumen. The composition addresses amyloid-beta-induced oxidative stress, mitochondrial dysfunction, and neuroinflammatory disorders. The invention is used to reduce the response, synaptic protein loss, and tau hyperphosphorylation. The subject matter includes the in vitro production method of the compound.

The invention relates to the technical field of biotechnology, enzyme engineering and biological pharmacy, in particular to a ribonuclease purification production method. According to the method, a deep sea antifreeze polypeptide and rare earth nanocrystal composite bionic system is constructed, quorum sensing signal molecules and a pulsed electric field are introduced in a fermentation stage for two-factor regulation, and a light-operated micro-fluidic chip selective extraction technology is combined, so that efficient separation of target enzyme is realized. A double-ligand magnetic microsphere chromatographic column is innovatively adopted for gradient elution, and the enzyme activity retention rate and the long-term storage stability are remarkably improved through thermotropic liquid crystal capsule stabilization and radio frequency assisted freeze drying processes. According to the invention, the destructiveness of the traditional chromatography technology to enzyme conformation is broken through, the problem of batch fluctuation caused by multi-step purification is solved, a ribonuclease product with high purity, high activity and environmental response characteristic is provided for the field of biological medicine, and the ribonuclease is especially suitable for industrial preparation of gene therapy products and virus vectors.

The invention discloses a recombinant vector containing a human astrovirus nucleic acid fragment, which is obtained by inserting an exogenous nucleotide sequence into a prokaryotic expression vector skeleton, and the exogenous nucleotide sequence is formed by sequentially connecting the following gene sequences: an SUMO fusion tag DNA sequence, an SAP gene sequence, an MS2 phage related DNA sequence and a human astrovirus sequence fragment. The tail end of the prepared human astrovirus pseudovirus particle contains a stem-loop structure, so that the human astrovirus pseudovirus particle is not easily degraded by nuclease, and the stability of virus nucleic acid RNA (Ribonucleic Acid) is improved; meanwhile, a human astrovirus conservative RNA gene sequence is wrapped by the phage MS2 coat protein, so that the phage MS2 coat protein has relatively high ribonuclease and deoxyribonuclease resistance, and the long-term stability of virus nucleic acid RNA is enhanced. And the stability problem of the virus RNA quality control product in the production and use processes is solved.

This disclosure provides several ribonuclease R variants and their preparation methods. Compared to the parent enzyme, the variants described in this disclosure exhibit increased protease activity per unit and show promising prospects for industrial applications. Furthermore, this disclosure also provides the application of ribonuclease R variants in linear RNA degradation.

The invention relates to gene therapy, and more specifically, to AAV gene therapy vectors containing a novel chimeric deoxyribonuclease (DNase) protein transgene and methods of treating ailments such as cancer and neurodegeneration.

The present disclosure provides compositions comprising sequence specific endoribonuclease and methods of their use in RNA analysis, RNA synthesis and fingerprinting of RNA molecules. In particular the present disclosure relates to compositions and samples comprising isolated endoribonuclease of Type III toxin-antitoxin systems preferably endoribonucleases of subfamily CptN and subfamily TenpN.

The invention relates to the technical field of biological medicine, in particular to DNase I loaded lipidosome and application thereof. The invention provides a bionic long-acting deoxyribonuclease I (DNase I) liposome with free DNA (cfDNA) responsiveness and a dual-targeting function, the liposome is subjected to layered fusion modification through an erythrocyte membrane and a neutrophile granulocyte membrane, a release window is accurately regulated and controlled by utilizing a 4ME-PC proportion, and a liposome drug carrier for cfDNA targeted combination and accurate delivery of an inflammation part is realized. The polypeptide can be used for cfDNA-related inflammatory diseases, and has an important application prospect.

The invention provides an application and a method for repairing neuronal dendritic growth defects by using ribonuclease RNase H. In particular, ribonuclease RNase H can be used as an R-loop degradation agent for treating and / or preventing neuron dendritic growth defects and related diseases thereof. By using a Rett syndrome cell model, it is found that the growth of dendrites can be restored by expressing ribonuclease RNase H of mammals. And a very significant means is provided for the treatment and prevention of Rett syndrome and other neuronal dendritic growth defect related diseases.

The invention discloses a ribosome group or selective ribosome group detection and analysis method based on an immune principle, and relates to the field of molecular biology and genomics. Comprising the following steps: sample preparation and cross-linking: treating cells by using a translation inhibitor to suspend ribosome translation, and then cross-linking RNA-protein in the cells by using a membrane permeable cross-linking agent; then cracking the cells, digesting RNA of a cracking product by using ribonuclease, and retaining ribosome and mRNA fragments protected by the ribosome; immune enrichment: carrying out specific immune enrichment on the digested sample by using an antibody combined with a carrier, and eluting to obtain a ribosome-mRNA compound; then decrosslinking and removing protein, extracting mRNA fragments and carrying out phosphorylation treatment so as to add joints, build a library and carry out sequencing; and finally, library construction, sequencing and bioinformatics analysis are carried out. The method has the advantages of high-specificity enrichment, accurate positioning of translation sites, wide application range and accurate sequence information.

The invention discloses an efficient ribonuclease double-primer isothermal amplification method which comprises a pair of ribonucleotide embedded hairpin primers, strand displacement DNA polymerase, ribonuclease, nucleic acid embedded dye and a target nucleic acid sequence. The ribonucleotide embedded hairpin primer is composed of a 5 '-end hairpin sequence, a 3'-end linear sequence and ribonucleotide embedded between the 5 '-end hairpin sequence and the 3'-end linear sequence, ribonuclease recognizes and cuts the ribonucleotide embedded in an amplification product and forms a gap, and the tail end of the gap can serve as a primer for further extension and replacement. According to the ribonuclease double-primer isothermal amplification method disclosed by the invention, an amplification product of which the 3 '-end sequence can be subjected to intramolecular hybridization in the amplification process can be autonomously extended and can be paired and extended by other primers as a substrate, and the ribonuclease double-primer isothermal amplification method disclosed by the invention only needs one pair of primers, has the advantages of simple primer design, high sensitivity and high specificity, and has a great application prospect.

The present disclosure provides, among other things, methods for removing linear ribonucleic acid molecules from a sample by contacting the sample with ribonuclease R (RNase R) from a thermophilic organism, for example, Geobacillus stearothermophilus, Fervidobacterium pennivorans or Thermus thermophilus expressed and purified in E.coli. Also provided herein are methods of producing circular RNA and removing uncircularized linear RNA molecules, linear RNA byproducts, or linear RNA side products using an RNase R expressed or purified from a thermophilic organism. The present disclosure also provides methods of purifying RNase R from a thermophilic organism in E. coll, wherein the method comprises heat treatment at high temperatures, and compositions and kits comprising the same.

The invention discloses a composite agent for preventing and treating rice blast, and belongs to the technical field of crop sterilization agents. The agent comprises a fungicide, an antiviral agent, a disease and pest resistant agent and a compound microbial agent according to a mass ratio. Wherein the fungicide is mainly formed by mixing iprodione, spinosad and cyprodinil, the antiviral agent is formed by mixing glucosamine, ribonuclease and beta-carotene, the disease and pest resistant agent is formed by mixing glyphosate and glufosinate-ammonium, and the compound microbial agent is formed by compounding beauveria bassiana and beauveria bassiana. Through reasonable matching of the components, the synergistic effect is achieved, the resistance of the components is reduced, and the pesticide effect is improved. In addition, the composite agent not only can effectively control the occurrence of rice blast, but also can improve the resistance of plants to the rice blast by enhancing the stress resistance and immune function of the plants, thereby achieving the purpose of prevention and treatment.

The invention relates to the technical field of gene engineering, in particular to a fusion protein expression cassette for rice multi-gene editing and application of the fusion protein expression cassette. The expression of the fusion protein for multiple gene editing of rice provided by the invention comprises an endonuclease Csy4, a DNA (Deoxyribose Nucleic Acid) endonuclease nickase Cas9, an M-MLV reverse transcriptase and a Cas12a nuclease; in the guide editing fusion protein frame, the ribonuclease Csy4 is located at the N end of the guide editing fusion protein frame, the DNA incision enzyme nickase Cas9 is located between the ribonuclease Csy4 and the M-MLV reverse transcriptase, the M-MLV reverse transcriptase is located between the DNA incision enzyme nickase Cas9 and the Cas12a nuclease, and the Cas12a nuclease is located at the C end of the fusion protein frame. The fusion protein expression cassette not only can ensure the editing efficiency, but also can improve the editing accuracy.

The invention provides a nucleic acid isothermal amplification detection system and a detection method thereof, which are characterized in that a primer containing a cleavage recognition sequence of incision endonuclease and a probe containing at least one ribonucleotide base are designed through combined use of incision endonuclease, DNA polymerase (reverse transcriptase is added for an RNA template) and ribonucleic acid endonuclease, and the nucleic acid isothermal amplification detection system is obtained. The primers and the probes are subjected to basic group modification, so that the signal intensity of the target nucleic acid is continuously amplified in the isothermal amplification process, the detection sensitivity is remarkable, meanwhile, mismatching can be avoided, non-characteristic amplification is reduced, the detection specificity is improved, missing detection and wrong detection are prevented, the detection time can be shortened, and the application range can be widened.

Methods and compositions for the detection and quantification of nucleic acids are provided. In certain embodiments, methods involve the use of cleavable probes that comprise a ribonucleotide position that is susceptible to endoribonuclease (e.g., RNase H) cleavage in the presence of target nucleic acid molecules. Probes of the embodiments may also comprise non-natural nucleotide linked to a reporter and / or quenching moiety.

A dental paste or gel is provided comprising a composition having: at least 2 to 4 % w / w agar; at least 62 to 95 % w / w glycerol; and at least 0.03 to 3 % w / w epithelial-adhering protein complex. The e