68 results about "Ribonuclease" patented technology

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 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 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.

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.

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.

The purpose of the present invention is to provide an immunogenic composition containing a new HBV polypeptide or polynucleotides encoding the polypeptide. The immunogenic composition contains at least one polypeptide selected from among polypeptides a) through c) or polynucleotides encoding the polypeptide, and can be used for treating or preventing HBV infection. a) an HBV surface protein polypeptide including a specific amino acid substitution; b) an HBV polymerase polypeptide including an inactivated reverse transcriptase domain and an inactivated ribonuclease H domain, and including a specific amino acid substitution; and c) an HBV core protein polypeptide including a specific amino acid sequence.

The invention relates to a novel liposome polydopamine nanoparticle and application thereof in treating renal cell carcinoma. The preparation method of the nanoparticles comprises the following steps: adding liposome into si-circPVT1 dissolved in ribonuclease-free water, and carrying out vortex mixing to form a si-circPVT1 liposome suspension; dispersing the dopamine PDA modified liposome into a Tris-HCl solution, then adding dopamine hydrochloride, and stirring to obtain dopamine PDA modified liposome; then dispersing in a streptavidin solution, and fully oscillating at 4 DEG C under a dark condition to obtain a compound; and mixing the complex with a biotin-labeled MUC12 antibody solution for incubation, and washing the obtained suspension with a PBS (Phosphate Buffer Solution) to obtain the novel liposome polydopamine nanoparticles marked as SCLPM-NPs. The product prepared by the invention realizes targeted delivery of kidney cell cancer cell lines, and the SCLPM-NPs can effectively target tumor tissue parts and inhibit the growth of kidney tumors.

The invention relates to the technical field of molecular biology, in particular to a structural probe and application thereof. The invention provides a nucleic acid probe which is single-stranded oligonucleotide, and the single-stranded oligonucleotide has a stem-loop structure; the number of the stem rings is larger than or equal to 2. The detection method provided by the invention is high in sensitivity, has ultrahigh sensitivity higher than that of a common single-chain fluorescent probe as an RNase reaction substrate, and can detect RNase A as low as 0.1 pg / ml. The probe disclosed by the invention has broad spectrum and has detection capability on various ribonuclease.

Compositions and methods relating to ApoA-1 fusion polypeptides are disclosed. The fusion polypeptides include a first polypeptide segment corresponding to an ApoA-1 polypeptide or ApoA-1 mimetic, and may also include a dimerizing domain such as, e.g., an Fc region, which is typically linked carboxyl-terminal to the first polypeptide segment via a flexible linker. In some embodiments, the fusion polypeptide further includes a second polypeptide segment located carboxyl-terminal to the first polypeptide segment and which confers a second biological activity (e.g., an RNase, paraoxonase, platelet-activating factor acetylhydrolase, cholesterol ester transfer protein, lecithin-cholesterol acyltransferase, polypeptide that specifically binds to proprotein convertase subtilisin / kexin type 9, or polypeptide that specifically binds to amyloid beta). Also disclosed are dimeric proteins comprising first and second ApoA-1 fusion polypeptides as disclosed herein. The fusion polypeptides and dimeric proteins are useful in methods for therapy.

The invention discloses a plasmid extraction kit and an extraction method. The plasmid extraction kit comprises a resuspension solution, a lysis solution, a neutralizing solution, a cleaning solution and an eluent, the resuspension is a Tris-HCl aqueous solution containing EDTA (Ethylene Diamine Tetraacetic Acid), glucose, ribonuclease, lysozyme and polyethylene glycol hexadecyl ether. The invention also discloses a plasmid extraction method which comprises the following steps: centrifuging a bacterial liquid of gram-negative bacteria containing plasmids, collecting thalli, and discarding a culture medium; re-suspending: adding a re-suspending solution, fully re-suspending, uniformly mixing, and incubating; sequentially carrying out steps of splitting decomposition, neutralization, purification, cleaning, centrifugal idling and elution to obtain a clarified solution containing plasmids. According to the method disclosed by the invention, the polyethylene glycol hexadecyl ether is added into the resuspension, and incubation is performed, so that the plasmid extraction amount is greatly increased, and the loss of plasmid extraction in subsequent links is reduced.

The present disclosure provides a plurality of RNase R variants and a preparation method thereof, wherein the expression amount of the variant RNase R is significantly increased relative to the parent enzyme, and the enzyme activity of the variant RNase R can be increased by 3 times, and the variant RNase R has a good industrial application prospect. In addition, the present disclosure also provides the application of the RNase R variant in the degradation of linear RNA and the preparation of circRNA.

This disclosure is directed to compounds and pharmaceutical compositions for treating and preventing viral diseases, as Covid-19. Among others, the invention relates to the use of immune cells and ribonucleases in the preparation and use of pharmaceutical formulations for the treatment of said disease.

The invention relates to a guided editing system, in particular to pegRNA, a pegRNA combination, a vector for expressing the pegRNA or the pegRNA combination, a vector for expressing PE protein, a tool cell, a guided editing system, a guided editing method, a kit and application of the pegRNA, the pegRNA combination, the vector, the tool cell, the guided editing system, the guided editing method and the kit in preparation of drugs for treating genetic diseases. The pegRNA comprises a first Csy4 endo ribonuclease recognition site and a second Csy4 endo ribonuclease recognition site, and the first Csy4 endo ribonuclease recognition site and the second Csy4 endo ribonuclease recognition site are subjected to enzyme digestion by Csy4 endo ribonuclease. According to the invention, high editing efficiency can be maintained between different target sites and cell types, and large-fragment DNA can be inserted and / or seamless editing can be realized.

The present application belongs to the technical field of genetic engineering, and particularly relates to a method for preparing recombinant deoxyribonuclease by prokaryotic expression. The method comprises the following steps: S1, connecting a target gene into a multiple cloning site of an expression vector to construct a recombinant expression vector, wherein the target gene comprises an X X A tag coding gene and a deoxyribonuclease coding gene; S2, introducing the recombinant expression vector into E. coli; S3, culturing the recombinant E. coli and inducing expression; and S4, collecting bacterial bodies after centrifugation of the recombinant E. coli culture solution, ultrasonic crushing, centrifugal collection of cell crushing supernatant, and then affinity purification to obtain a deoxyribonuclease fusion protein. The present application fuses the X X A tag with the deoxyribonuclease, adopts a prokaryotic expression system, has high host bacterium activity in the process of inducing expression, realizes high-efficiency expression of DNase, and significantly improves the soluble expression level, and has the advantages of good DNase biological activity and high stability.

The invention provides an integrated DNA interference excluding in-situ hybridization probe and an in-situ hybridization method for detecting hepatitis B virus cccDNA, the in-situ hybridization probe is designed based on a Basescope detection method, and comprises an HBV cccDNA probe specifically targeting an integrated breakpoint of HBV negative chain DNA, an HBV DNA probe targeting an HBV negative chain, an HBV cccDNA probe targeting an HBV negative chain, an HBV cccDNA probe targeting an HBV negative chain, an HBV cccDNA probe targeting an HBV negative chain, and an HBV cccDNA probe targeting an HBV negative chain. The HBV RNA probe with the targeted HBV positive strand gap structure and the matched ribonuclease of the HBV RNA probe can be used for detecting cccDNA and integrating HBV DNA after being treated, and the targeting sequences of the HBV cccDNA, the HBV DNA and the HBV RNA probe are respectively shown as SEQ ID No.1 to SEQ ID No.3. The in-situ hybridization probe and the in-situ hybridization method can be applied to screening medicines for preventing or treating hepatitis B virus infection and researching the life cycle of the hepatitis B virus, so that the understanding on the characteristics of the hepatitis B virus is promoted, and the evaluation on a treatment strategy is optimized.

The present application relates to a chimeric primer-mediated nucleic acid detection method and detection kit. The detection method provided by the present invention mainly introduces a chimeric primer into a constant temperature amplification system, degrades the RNA portion of the hybrid DNA double strand formed by amplification through ribonuclease, and highlights the RNA binding site of the chimeric primer. Subsequently, the protruding gap produced by ribonuclease degradation of RNA is used to achieve exponential amplification of the target gene under the action of the strong chain displacement activity of the strand displacement DNA polymerase. The entire amplification process does not require high-temperature denaturation of the nucleic acid, is simple to operate, and is low in cost.