42 results about "Integrase" patented technology

Among other things, provided herein are systems that replace the natural random integration activity of a retrovirus with site-specific integration machinery. This approach allows for a more precise targeting of a gene of interest into a human genome, e.g., for therapeutic purposes. The system may include integration-deficient retrovirus (e.g., lentivirus) (IDLV), in which the natural integration activity has been reduced (e.g., by mutation to the viral integrase polypeptide). Instead, the system may comprise a site-specific recombinase (e.g., a serine recombinase, e.g., a serine integrase) capable of directing insertion of a template DNA, or portion thereof, into a desired site in the human genome.

The invention discloses a mycobacteriophage and application thereof, and relates to the technical field of bacteriophages. The mycobacteriophage is named as Mycobacter phage BZNK001, is preserved in Guangdong Microbial Culture Collection Center, has a preservation number of GDMCC No: 67907-B1, does not contain antibiotic drug-resistant genes and integrase genes, has excellent biological safety and good environmental tolerance, can maintain high biological activity in a temperature range of 37-50 DEG C under an acid-base condition of pH 5-9, and can be used for preparing the antibiotic drug-resistant gene and integrase gene-containing mycobacteriophage for preventing and treating the antibiotic drug-resistant gene and integrase gene-containing mycobacteriophage for preventing and treating the antibiotic drug-resistant gene and integrase gene-containing mycobacteriophage. The bacillus subtilis has broad-spectrum and high-efficiency non-tuberculous mycobacterium cracking activity, and can effectively crack four clinical key mycobacteria such as mycobacterium abscessus, mycobacterium avium, mycobacterium intracellularis and mycobacterium smegmatis. According to the invention, a mycobacteriophage resource library is enriched, a novel and stable biological prevention and control means is provided for efficient prevention and control of mycobacteria, and the mycobacteriophage has good clinical application and industrialization values.

The invention provides a method for synchronously and rapidly detecting Escherichia coli and multiple drug-resistant genes thereof, which comprises the following steps: culturing a sample at night to obtain a bacterial solution to be detected, adding a buffer solution and a metal bath, centrifuging, taking a supernatant, detecting by micro-fluidic, optimizing the extraction process of nucleic acid in the sample, designing an optimal RPA primer probe group, and detecting the multiple drug-resistant genes of the Escherichia coli. A reasonable RPA coupled microfluidic detection system is established, synchronous and rapid detection of the escherichia coli uidA and I-type integrase gene intI1 thereof and drug-resistant genes sul1 and aadA5 is realized, the specificity is high, the sensitivity is high, the reaction time is short, the operation is convenient, and a method support is provided for monitoring and prevention and control of food-borne drug-resistant bacteria.

A non-integrating viral delivery system is disclosed. The system includes a viral carrier, wherein the viral carrier contains a defective integrase gene; a heterologous viral episomal origin of replication; a sequence encoding at least one initiator protein specific for the heterologous viral episomal origin of replication, wherein expression of the sequence encoding the at least one initiator protein specific for the heterologous viral episomal origin of DNA replication is inducible; and at least one gene, gene product, shRNA, siRNA, miRNA, or other RNA of interest.

The construct for site-specific insertion of DNA fragments and transgenes into the genome comprises a recombinant enzyme. This enzyme contains a modified HIV-1 integrase (HIV IN) tetramer with multiple mutations favoring a heterodimer structure in which the first and second outer monomers are bound to the first and second inner monomers, respectively. A transcription activator-like effector (TALE) is bound to the N-terminal region of the outer monomer via a polypeptide linker. The two TALEs confer sequence specificity to the enzyme and prevent non-specific interactions with the target DNA structure. The modified HIV IN monomer ensures that the TALE binds to the outermost monomer and that the TALE is positioned relative to the catalytic region of HIV IN. A polynucleotide is bound to the inner monomer of HIV IN. When the TALE is transported to the nucleus, either alone or within a lentiviral capsid, it interacts with the DNA to localize HIV IN, which then incorporates the polynucleotide.

The invention discloses a test strip biosensor for detecting uranyl ions, the test strip biosensor comprises a reaction system and a test strip, the reaction system comprises a DNAzyme substrate chain and a DNAzyme enzyme chain; the test strip comprises a substrate, a sample pad, a coupling pad, a nitrocellulose membrane and an absorption pad, the sample pad, the coupling pad, the nitrocellulose membrane and the absorption pad are sequentially overlapped on the substrate, a gold nanoparticle-DNA probe I compound is fixed on the coupling pad, a test line and a control line are arranged on the nitrocellulose membrane, an SA-biotin-DNA probe II is fixed on the test line, and an SA-biotin-DNA probe II is fixed on the control line. And an SA-biotin-DNA probe III is fixed on the control line. Compared with the prior art, the biosensor can realize high-sensitivity and high-specificity detection of uranyl ions by integrating a DNA enzyme induced cleavage reaction and a gold nanoparticle mediated colorimetric detection technology, the detection process is simple and rapid, and an important technical support is provided for field monitoring of UO2 < 2 + > in environment and biological samples.

The present disclosure provides a pyridone compound having integrase inhibitory activity and a pharmaceutical use thereof. Specifically, the present disclosure provides a compound having a structure shown as formula I or a pharmaceutically acceptable salt thereof, which can be used to treat human immunodeficiency (HIV) infection.

The present disclosure relates to the field of biotechnology, and particularly relates to detection of BXB1 integrase. The present disclosure establishes a method for detecting BXB1 integrase based on double-antibody sandwich ELISA technology, which has good sensitivity and specificity, and the method is simple to operate and can quickly realize accurate quantification of BXB1 integrase; in addition, the present disclosure also provides an ELISA kit for detecting BXB1 integrase. By using the BXB1 integrase detection method or kit provided by the present disclosure, accurate quantification of the residual amount of BXB1 integrase in various intermediate products, semi-finished products or finished products in the process of preparing recombinant biological products by the BXB1 integrase system can be realized, which is beneficial to the quality control of products by biopharmaceutical enterprises.

The invention discloses an enzymatic colorimetric coding-decoding system for weighted detection of multiple biomarkers and application, and belongs to the field of biomedical detection. According to the enzymatic colorimetric coding-decoding system provided by the invention, the mapping relation between the color and the multi-dimensional biomarker can be realized through direct visual interpretation or simple spectral analysis. The invention also provides an enzymatic colorimetric coding-decoding method for weighted detection of multiple biomarkers, and the method comprises the following steps: converting multi-dimensional biomarker information into quantifiable and visually analyzable optical signals through an integrase catalyzed multicolor reaction, nucleic acid driven signal amplification and optical coding-decoding mechanism; and single-tube multi-dimensional joint detection, high information density output and rapid disease state discrimination are realized. The method is successfully applied to early screening of pancreatic cancer and can be expanded to dynamic monitoring of multiple markers of other cancers and infectious diseases in the future, and an efficient and low-cost solution is provided for precise medical treatment and digital medicine.

The present invention relates to the technical field of drug for treatment of AIDS. Disclosed are a polycyclic carbamoyl pyridone derivative, a preparation method therefor, and pharmaceutical composition thereof. The derivative comprises the compound represented by formula (I)-1 or formula (I)-2, or an isomer, pharmaceutically acceptable salt, hydrate or solvate thereof. At least one group of functional groups in (1)-(3) below and a ring A form a spirolcycle or spiroheterocycle: (1) R1 and R2; (2) R3 and R4; (3) R5 and R6; the polycyclic carbamoyl pyridone derivative can selectively inhibit the activity of HIV integrase, and thus can be used for preventing and treating AIDS.

This invention discloses a biosensor for detecting uranyl ions using a test strip, comprising a reaction system and a test strip. The reaction system includes a DNAzyme substrate chain and a DNAzyme enzyme chain. The test strip includes a substrate and a sample pad, a coupling pad, a nitrocellulose membrane, and an absorbent pad sequentially overlapped on the substrate. A gold nanoparticle-DNA probe I complex is immobilized on the coupling pad. A test line and a control line are provided on the nitrocellulose membrane. An SA-biotin-DNA probe II is immobilized on the test line, and an SA-biotin-DNA probe III is immobilized on the control line. Compared with existing technologies, this biosensor integrates a DNase-induced cleavage reaction with gold nanoparticle-mediated colorimetric detection technology, achieving high sensitivity and high specificity detection of uranyl ions. Furthermore, the detection process is simple and rapid, providing a solution for detecting UO2 in environmental and biological samples. 2+ This provides important technical support for on-site monitoring.

The invention relates to a cyclic peptide comprising the amino acid sequence N-terminal-X4X3X2X1-a-Y1Y2Y3Y4-C-terminal, wherein -a- represents a linker, and wherein the peptide comprises a linker -c- between the N-terminal amino acid and C-terminal amino acid, and / or wherein the two amino acids of one or two of the following amino acid pairs form a side chain-to-side chain bridged di-amino acid forming a bridge -b-: X1 and X1; X2 and Y2; X3 and Y3; and X4 and Y4. The invention further relates to the use of said cyclic peptide as a medicament, especially for inhibiting or preventing a Wingless / integrase-1 (Wnt)-dependent disease in an individual in need thereof.

The application discloses streptomyces 891-DC with high aurovertin A yield and application thereof. The application uses CRISPR-Cas9 technology to first change an incomplete attB site sequence in the genome of streptomyces 891 into a complete attB site sequence, promote a plasmid containing a target gene chry, an integrase phiC31 gene and an attP site, and improve the integration rate of the target gene through specific combination of the attP site and the attB site on the genome. Secondly, a promoter capable of obviously improving the yield is screened to express the aurovertin biosynthesis gene cluster, so that the more stable and higher-yield streptomyces 891-DC is obtained. The aurovertin A shake flask fermentation yield is 1411.23+ / -29 mg / L, which is 60% higher than that of the original strain, and the genetic stability is good, thereby laying a technical foundation for industrial production of aurovertin.

The invention relates to the technical field of biology, in particular to a bacteriophage Ca1 and a preparation method thereof, the bacteriophage is the bacteriophage Ca1 capable of infecting a strain Xcc8004 and is preserved in Guangdong Microbiological Culture Collection Center, and the preservation number is GDMCC66226-B1. According to the phage Ca1 and the preparation method thereof disclosed by the invention, the highest average nucleotide similarity between the genome sequence of the phage Ca1 and the known phage sequence is 94% and is lower than a new species threshold (95%), and the genome does not contain integrase, so that the phage Ca1 is a new lytic phage; the titer is relatively stable in a pH range of 3-11; the titer of the bacteriophage is relatively stably maintained at 109 PFU / mL at 4-60 DEG C; after the bacteriophage Ca1 is treated by 25% chloroform, the titer is not obviously changed, which indicates that the bacteriophage is insensitive to chloroform and free of lipid coating; the results show that the bacteriophage Ca1 is a new bacteriophage resistant to acid, alkali, high temperature and organic solvent, and has strong stability in natural environment.

The present invention belongs to the field of detection technology, and specifically discloses a detection method, kit and application of HIV-1 drug resistance genes based on NGS. The detection method includes the steps of: S1, extracting RNA from an HIV-1 serum or plasma sample; S2, amplifying and enriching the drug resistance detection region: performing one-step RT-PCR to amplify the protease region, reverse transcriptase region and integrase region of the HIV-1 POL gene, and detecting the amplification product; S3, library construction: performing fragmentation, adapter addition and tag addition and synchronous enrichment operations on the amplification product in S2 to construct a sequencing library; S4, performing on-machine sequencing; S5, data analysis: according to the sequencing data in step S4, analyzing and obtaining the genotypes and drug resistance evaluations of the protease and reverse transcriptase regions and integrase regions of the HIV-1 POL gene. The present invention only requires one-step RT-PCR, and does not require two-round nested PCR to specifically enrich the drug resistance detection region of the HIV-1 POL gene.

The present invention relates to a system comprising a landing pad, a delivery nucleic acid and an integrating enzyme allowing selection of a specific gene of interest. The invention further relates to cells comprising the system, a method for providing a gene of interest and selecting a cell with a gene of interest, a landing pad nucleic acid, a delivery nucleic acid and a kit for providing and selecting a cell with a gene of interest.

In the present invention, there is provided a nucleic acid molecule that is capable of recognizing and / or binding to HIV, in particular to the integrase (IN) of HIV-1 and HIV-2 variants or portions thereof. In particular, the nucleic acid molecule is capable of binding to the HCINp1 (11 aa) (SEQ ID NO 4, PYNPQSQGVVE), HCINp2 (9 aa) (SEQ ID NO 5, NFKRKGGIG), HCINp3 (9 aa) (SEQ ID NO 6, LLWKGEGAV), HCINp4 (8 aa) (SEQ ID NO 7, IKVVPRRK) and / or HCINp5 (10aa) (SEQ ID NO 15, LLWKGEGAVV). The nucleic acid molecule may be a polynucleotide or an oligonucleotide.

The invention provides compositions, systems, and methods for revising nucleic acids. The revision may be accomplished using a fusion protein comprising a nuclease, a ligase, an integrase, or a combination thereof. The nucleic acid revision may include ligation of a donor nucleic acid to a target nucleic acid. The nucleic acid editing may include replacement of a portion of the target nucleic acid with a revising nucleic acid.

The application discloses a corynebacterium striatum phage and application thereof. The corynebacterium striatum phage is a corynebacterium striatum phage CSP1, the preservation number of which is CGMCC NO: 45598, and the preservation time is July 4, 2023. The corynebacterium striatum phage has strong tolerance, biological safety and carries site-specific recombination functional elements (integrase and recombination sites). Meanwhile, the corynebacterium striatum phage can lyse 20 strains of multi-drug resistant corynebacterium striatum, and lays a foundation for subsequent medical researches using the corynebacterium striatum phage as a test and treatment carrier, a targeted pathogenic bacterium or symbiotic bacterium, and an immune regulator inducer.

The invention provides methods for genetically engineering T cells in vivo comprising administering to the subject a cytokine or nucleic acid molecule encoding a cytokine to recruit the subject's T cells to the administration site; followed by administration of a nucleic acid molecule encoding an antigen receptor. In some instances, the method also includes administering an integrase or nucleic acid molecule encoding an integrase to integrate the sequence encoding the antigen receptor into the DNA of the recruited T cells.

The invention relates to the technical field of gene engineering, in particular to an HIV-1 integrase drug resistance detection primer combination based on a next-generation sequencing technology and application and a kit of the HIV-1 integrase drug resistance detection primer combination. The primer combination comprises any one of an IN-Mix-PLA primer and a deep sequencing primer combination. The HIV-1 integrase drug resistance detection method established by using the primer combination provided by the invention has the advantage that the HIV drug resistance detection cost is greatly reduced. A single sample can produce more than 1500 sequences, so that sufficient data resources are provided for analyzing HIV-1 dominant drug-resistant strains and inferior drug-resistant strains, and powerful guidance can be provided for subsequent antiviral treatment. Therefore, HIV-1 drug resistance detection becomes a low-cost and high-yield method, and the method has great influence on AIDS prevention and public health.

The application discloses an enzymatic colorimetric encoding-decoding system and application for multiple biomarker weighted detection, and belongs to the field of biomedical detection. The enzymatic colorimetric encoding-decoding system can realize the mapping relationship between color and multidimensional biomarkers through direct visual interpretation or simple spectrum analysis. The application further provides an enzymatic colorimetric encoding-decoding method for multiple biomarker weighted detection. By integrating enzyme-catalyzed multicolor reaction, nucleic acid-driven signal amplification and optical encoding-decoding mechanism, the multidimensional biomarker information is converted into quantifiable and visually analyzable optical signals, realizing single-tube multidimensional joint detection, high information density output and rapid disease state discrimination. The application has been successfully applied to early screening of pancreatic cancer, and can be extended to dynamic monitoring of multiple markers of other cancers and infectious diseases in the future, providing an efficient and low-cost solution for precision medicine and digital medicine.

The present invention provides a method, system and application for large-fragment DNA cascade assembly, belonging to the field of biotechnology. The present invention provides a method and system for large-fragment DNA cascade assembly. Based on the integrase system-based bacterial chromosome assembly technology, by using two sets of orthogonal serine integrase systems, the present invention can efficiently assemble large-fragment DNA into ultra-long DNA in sequence; this method can perform cascade assembly of DNA in Escherichia coli, especially can assemble large-fragment DNA of more than 100 kb into ultra-long DNA through multiple rounds of assembly, and has important application values in aspects such as DNA assembly, large-scale genomic modification, synthesis of artificial chromosomes and construction of artificial living organisms.

The present invention relates to a composition of lentiviral particles comprising a mutated integrase and an RNA-guided nuclease, for targeted gene insertion.

The present invention relates to a V. cholerae strain in which a platform has been built to detect integron cassettes. The platform includes a blind reporter system built by inserting an attl integration site into the gene of the toxin of a toxin / antitoxin system, which is inserted in the chromosome of the bacterium; the gene of the corresponding antitoxin and the gene of the intl integrase, both included in plasmids. The invention also relates to V. cholerae strains in which the superintegron (ASI) has been deleted, to the method for obtaining said strains, and to V. cholerae ASI strains in which the platform for capturing integron cassettes has been included. The invention includes methods and kits for detecting integron cassettes by using the strains and platforms built.

The application discloses a kind of computing methods for identifying amino acid candidate mutation sites of enzyme, and the computing method includes enzyme protein hinge positioning, residue evolutionary conservation analysis and energy calculation.The application first adopts the method of integrating enzyme protein hinge positioning, residue evolutionary conservation analysis and energy calculation to screen enzyme candidate mutation sites, realizes the high-precision rapid screening of glycosyltransferase mutation sites, significantly improves the enzyme activity of glycosyltransferase on rebaudioside D (RebD) to produce rebaudioside M (RebM), effectively solves the problems of poor substrate specificity and low catalytic activity of the enzyme, reduces production cost, improves the soluble expression capacity of glycosyltransferase, is conducive to large-scale production through microbial fermentation, and is more suitable for commercial and industrial applications.

The present invention relates to a V. cholerae strain in which a platform has been built to detect integron cassettes. The platform includes a blind reporter system built by inserting an attl integration site into the gene of the toxin of a toxin / antitoxin system, which is inserted in the chromosome of the bacterium; the gene of the corresponding antitoxin and the gene of the intl integrase, both included in plasmids. The invention also relates to V. cholerae strains in which the superintegron (ASI) has been deleted, to the method for obtaining said strains, and to V. cholerae ASI strains in which the platform for capturing integron cassettes has been included. The invention includes methods and kits for detecting integron cassettes by using the strains and platforms built.

The invention relates to a tool for capturing integron cassettes, which includes a gene of a counterselective marker into which an integration site (attI) has been inserted to convert same into a blind reading marker for selecting recombination events. The tool also includes the integrase gene intI. The structure of the counterselective marker with the attI site can be integrated into the chromosome of a bacterium, especially of a strain of Vibrio cholerae without superintegron, while the intI gene is included in a plasmid in which the tfoX gene can also be inserted, the tfoX gene regulating the natural competence of V. cholerae. The invention also relates to methods and kits for detecting integron cassettes present in other bacteria by means of conjugation, and integron cassettes from DNA samples by means of natural transformation.

A herpes simplex virus (HSV) gene vaccine and expression vector and / or vaccine vector comprising an HSV genome comprising: a complete deletion of glycoprotein G-encoding gene, glycoprotein J-encoding gene, glycoprotein D-encoding gene, and C glycoprotein I-encoding gene; a heterologous nucleic acid comprising an expression cassette and inserted in a region of the genome from which the glycoprotein G-encoding gene, glycoprotein J-encoding gene, glycoprotein D-encoding gene, and glycoprotein I-encoding gene have been deleted; an attL sequence; and an attR sequence, wherein the attL sequence is adjacent to a first end of the expression cassette and the attR sequence is adjacent to a second end of the expression cassette, and wherein the expression cassette comprises in operable communication a promoter and at least one gene encoding a heterologous protein.