93 results about "Integrase" patented technology

Compounds of formula I:

wherein R₁, R₂, R₄, R₁₀, R₁₁, and Q are as defined herein, and their pharmaceutically acceptable salts, are useful in the prevention or treatment of HIV infections.

The subject invention provides a unidirectional site-specific integration system for integrating a nucleic acid into the genome of a target cell. The provided systems include a (1) a mutant, unidirectional site specific integrase, which can be provided by an integrase vector encoding the mutant integrase and (2) a targeting vector that includes: (a) a nucleic acid to be integrated; and (b) a vector attachment site, where the targeting vector attachment site serves as a substrate for the mutant, unidirectional site-specific integrase. In using the subject systems for site-specific integration, the targeting vector and integrase are introduced into the target cell and the cell is maintained under conditions sufficient to provide for site-specific integration of the nucleic acid into the target cell genome via a recombination event mediated by the site-specific integrase. Also provided are kits that include the subject systems. The subjects systems, methods and kits find use in a variety of different applications, several representative ones of which are described in detail as well.

The invention discloses an integrative plasmid pOPHI and a resistance screening marker-free self-luminescent mycobacterium. The integrative plasmid pOPHI comprises a promoter, an enzyme gene (LuxCDABE) needed for luminescence, a fusion gene of an integrase gene (Int) and a resistance screening gene, direct repeat sequences DifR and DifL positioned on two ends of the fusion gene, a phage integration site (attP) and a replicon. After the integrative plasmid pOPHI is transferred into the mycobacteria, a photobacterium of which the resistance gene and the integrase gene are lost is screened out by subculturing, and the photobacterium is the resistance screening marker-free self-luminescent mycobacterium. Whether the resistance screening marker-free self-luminescent mycobacterium obtained by construction is dead or not is judged according to whether the bacterium is luminescent or not, the mycobacterium can be used for a plurality of detections of experiments, and credible experimental data can be quickly obtained.

A compound of the formula (I) wherein Z<4>, Z<5> and Z<9> each is independently carbon atom or nitrogen atom; Y is hydroxy, mercapto or amino; R is a group of the formula (II) (wherein C ring is nitrogen-containing heteroaryl) has an inhibitory activity against integrase.

The invention discloses a method of constructing selection-marker-free self-luminescent mycobacterium abscessus and establishment of a corresponding in-vitro high-flux medicine-screening model. During the construction process, a key factor is the construction of a recombinant plasmid pOPAI1B, which contains a fused gene including a luminescent required enzyme gene LuxCDABE, a resistance screening gene Apr and an integrase gene Int; direct repeat sequences DifR and DifL at the two ends of the fused gene respectively; and a phage integration site attP and a replicon. The recombinant plasmid pOPAI1B is transformed into the mycobacterium abscessus and then is screened to obtain the selection-marker-free self-luminescent mycobacterium abscessus of which a resistance gene and the integrase gene are both lost. The self-luminescent mycobacterium abscessus can screen drugs in a high flux, can determine sterilizing or bacteria-resistant effects of the drug by means of detection of luminescent situation of bacteria, can be used for in-vitro drug activity detection, is free of any substrates, can continuously detect the same sample, is high in sensitivity and is strong in repeatability.

An embodiment of a method for detecting low frequency occurrence of one or more HIV sequence variants associated with integrase is described that comprises the steps of: (a) generating a cDNA species from a plurality of RNA molecules in an HIV sample population; (b) amplifying a plurality of first amplicons from the cDNA species, wherein each first amplicon is amplified with a pair of nucleic acid primers; (c) clonally amplifying the amplified copies of the first amplicons to produce a plurality of second amplicons; (d) determining a nucleic acid sequence composition of the second amplicons; (e) detecting one or more sequence variants that occur at a frequency of 5% or less in the nucleic acid sequence composition of the second amplicons; and (f) correlating the detected sequence variants with variation associated with HIV integrase.

The invention relates to a nitrogen-containing polyhydroxy arometic compound having a structure as shown in a general formula 1, wherein R1 is hydrogen, methyl or fluorine; R2 is hydrogen, methyl, acetyl, fluorine, chlorine or bromine; R3 is hydrogen, methyl, butyl, methoxyl, ethoxyl, fluorine, chlorine, bromine or nitro; R4 is hydrogen, methyl, acetyl, fluorine, chlorine or bromine; and n is 0 or 1. The invention also relates to a method for preparing the nitrogen-containing polyhydroxy arometic compound in the general formula 1. The compound has obvious inhibitory activity to HIV integrase, and is used for preparing anti-AIDS medicine. The n is equal to 0 or 1.

The invention discloses a preparation method of a pseudotype virus for nucleic acid detection of 2019-nCoV, and relates to the technical field of biology. The preparation method comprises a method forpreparing standard quality control products which contain but are not limited to a 2019-nCoV nucleotide sequence and are used for nucleic acid detection through a non-integrated slow virus vector system, a method which is used for preparing the standard quality control products for nucleic acid detection, and a method for constructing a non-integrated slow virus through mutating 64-locus amino acid Asp of integrase of slow virus auxiliary plasmid into Asn, and amino acids Arg, Arg and Lys at 262-264 loci into Ala, Ala and His. In the manner of removing elements of promoters and the like of slow viruses, the load quantity of virus vectors can be promoted, a pseudotype virus which can be used for nucleic acid detection and complete flow quality control is constructed, and the preparation method can be used for constructing a pseudotype virus for corona viruses and can also be used for constructing a pseudotype virus for SARS-COV, MERS-COV, influenza viruses and the like.

The invention discloses a compound type piggyBac recombinant vector as well as a preparation method and an application of the compound type piggyBac recombinant vector. According to the recombinant vector, a target gene expression frame, selection marker gene expression frames I and II, piggyBac transposase expression frame regulated by a heat shock promoter and truncated type piggyBac transposon arms L2 and R2 are inserted between wild type piggyBac transposon arms R1 and L1, integration of target gene, selection marker gene, piggyBac transposase expression frame sequence in silkworm genome can be conveniently mediated, complete deletion of all of transposon sequences and selection marker gene sequences in progeny transgenosis silkworm genome can be realized through establishing and screening a transgenosis silkworm line with high target gene expression and then inducing piggyBac to express in the transgenosis silkworm by heat shock treatment, replacement of other exogenous gene and the target gene can be conveniently realized through box type exchange reaction mediated by phiC31 integrase by surplus target gene expression frame anchored by attP locus, and then fixed point integration of other exogenous genes in the genome locus is further realized.

The invention belongs to the technical field of biological engineering and synthetic biology, and particularly relates to a multi-fragment deoxyribose nucleic acid (DNA) series connection recombination assembly method based on BT1 integrase and mutation recognition sites thereof. In the method, sixteen pairs of mutational integrase recognition sequences are provided through mutation screening and verification, on the basis, a complete set of related carriers is constructed and comprises a framework carrier and seven TA cloning vectors, the framework carrier can realize escherichia coli-streptomyces coelicolor shuttle, the seven TA cloning vectors comprise mutation substrates and are used for DNA element construction and screening, and a concrete method for multi-fragment external series connection recombination reaction is provided. The method of the invention is simple, fast and efficient and can be used for the multi-fragment fast series connection recombination assembly, and the amplification of series connection products is realized through escherichia coli transformation. The method can be widely used for combination and assembly of standard modules and elements in the synthetic biology.

Method for separating and determining bovine genome pseudo attp site and its usage are disclosed. The process is carried out by separating out tow pseudo attp sites from bovine gene, having 38% and 41% homology with attp site of streptomycete phagicin phiC31 genome, discriminating by streptomycete phagicin phiC31 integrase and integrating foreign gene containing attB site with the site of bovine geneome under mediation of integrase. It has higher integration efficiency.

The invention relates to (Z)-1-(1-substituted benzyl-5-methyl-1H-1,2,3-triazole-4-yl)-3-substituted benzyl-3-hydroxy-2-propylene-1-ketone compound and a preparation method and an application thereof. The compound is expressed by formula (I), wherein R1 represents -F, R2 represents -H, or R1 represents -H, R2 represents -F; R3, R4 and R5 represent -OH or -H. The preparation method of the compound comprises synthesizing fluoro-benzyl azide with mono-fluoro benzyl bromide as raw material, reacting with acetylacetone to obtain 1-(1-fluoro-benzyl)-4-acetyl-5-methyl)-1H-1,2,3-triazole, carrying out condensation reaction with substituted methyl benzoate to obtain a compound having a dicarboxylic acid structure, and finally demethylating with boron tribromide to obtain the compound. The compound provided by the invention has an inhibitory action to HIV-1 (human immunodeficiency virus-1) integrase.

The invention belongs to the technical field of medicines and specifically provides a multi-substituted amine compound as shown in a general formula I or an isomer thereof or a pharmaceutically acceptable salt, ester, pro-medicament or hydrate thereof or a pharmaceutical composition, as well as a preparation method and use thereof in preparation of medicaments for treating acquired immune deficiency syndrome. The compound or the pharmaceutical composition containing the compound can be used as an inhibitor for inhibiting the protein-protein interaction between HIV (human immunodeficiency virus) integrase and LEDGF/p75 and the dimerization of the HIV integrase, and further can be used for treating the acquired immune deficiency syndrome.

The invention provides a novel integrase and application thereof to genetically modifying saccharopolyspora spinosa. The integrase particularly comes from saccharopolyspora endophytica CCTCC AA208003, and exogenous DNA (deoxyribonucleic acid) can be specifically locally integrated on saccharopolyspora endophytica chromosome by the integrase via specific cis-form components. In addition, independent free replication regions of different strain sources are discovered in the saccharopolyspora spinosa by an inventor via research on genetic factors of different strains of the saccharopolyspora spinosa. The novel integrase and the application have the advantages that vectors constructed on the basis of the integrase and sequences of the replication regions can be used for genetically modifying the saccharopolyspora spinosa, accordingly, novel spinosad analogues can be possibly discovered, and the spinosad fermentation yields can be possibly increased; different resistance molecular markers can be carried by plasmids, accordingly, the novel integrase can be used for inter-species or intergenetic genome shuffling operation on the saccharopolyspora spinosa, and efficient genome shuffling can be implemented.

The invention relates to the field of molecular diagnostic, in particular for the detection of the presence of gram-negative bacterial strains resistant to antibiotic in a biological sample. The invention more specifically relates to an in vitro method for detecting the presence of gram-negative bacterial strains resistant to antibiotics in a biological sample, said method comprising the steps of: a) providing a biological sample; b) preparing said biological sample for nucleic acid amplification; c) performing nucleic acid amplification using (i) nucleic acid from said biological sample as a template, (ii) at least one or more set of primers specific of bacterial genes encoding integrase of integrons of class 1, 2 and 3, and, (iii) at least one or more set of primers specific of bacterial genes encoding CTX-M type β-lactamases; and, d) determining the presence or absence of amplicons; wherein the presence of at least one amplicon is indicative of a high likelihood that said biological sample contains bacterial strains resistant to antibiotics. The method may be carried out directly on clinical samples, e.g. from septic patients.

The invention discloses pyrrolopyridazine compounds used for treating or preventing HIV (human immunodeficiency virus) infection or other virus infections. The compounds can be used for as inhibitors for inhibiting HIV replication and can be used with other treatment drugs (particularly other antiviral drugs). Specifically, the invention discloses compounds for inhibiting HIV replication, a drug containing the compounds and a method for treating retrovirus infection through the compounds, for example, HIV infection. More specifically, the invention discloses an integrase allosteric inhibitor.

The invention belongs to the technical fields of biotechnology and gene engineering, and in particular relates to a method for quickly constructing a gene targeting vector based on phi BT1 integrase and a mutation recognition site thereof. The invention provides three pairs of mutative intergrase recognition sequences by strict mutation screening and verification, and the mutative substrate pair has equal reaction efficiency to that of a wild type, and appears as incompatible in the same system. On the basis, the method constructs a set of related vectors comprising a backbone vector capable of realizing Escherichia coli-streptomyces coelicolor shuttle, and three TA clone vectors containing mutative substrate pairs for constructing and screening homology arms, and provides a concrete method of series recombination and reaction of four DNA fragments in vitro. The method of the invention is simple, quick and high-efficiency, can be used for constructing the gene targeting vector of streptomyces, and can be widely used for quickly constructing gene targeting vectors of different species by corresponding original substitution.

The present invention relates to agents based on integrase of HIV-1, for inhibiting the proliferation of HIV-1. The agents are derived from the C-terminal domain of HIV-1 integrase, comprising at least one of the regions identified as being important for interaction between integrase and imp7 or impβ, and/or for nuclear localization of the HIV PIC, replication of HIV, or infection of HIV.

The invention discloses a mouse model capable of monitoring NF (nuclear factor)-kB activity in the liver by virtue of in-vivo imaging and a construction method of the mouse model. The construction method comprises the following steps: constructing a recombinant vector which carries a phage integrase recognition site attB gene, an NFkB gene and an Fluc (firefly luciferase) reporter gene, and introducing the recombinant vector into the mouse body by virtue of a hydrodynamic method to obtain the mouse model, wherein the Fluc reporter gene regulated and expressed by the NFkB gene is integrated in the liver of a mouse. The mouse model disclosed by the invention can be used for evaluating the adjustment effects of different factors on the NF-kB activity in vivo, also can be used as an evaluation model of interferon or inflammatory factor regulating medicaments, molecules and viruses in natural immunity, and can be widely applied to various categories of biomedical animal experiments.

The invention provides a degenerate primer set for one-time human immunodeficiency virus (HIV) genotypic resistance detection and application of the degenerate primer set, and relates to the technicalfield of virus detection. The degenerate primer set includes a reverse transcription PCR primer, a nest PCR amplification primer and a sequencing universal primer; the reverse transcription PCR primer includes WC-pol-F and WC-pol-R; the nest PCR amplification primer includes WC-nest-F and WC-nest-R; and the sequencing universal primer includes HBX2-3017R, HBX2-3785R and HBX2-4561R. The degenerateprimer set is applicable to genotypic resistance detection of most of HIV-1 strains prevalent in China, a gene coding region of HIV protease-everse transcriptase-integrase can be amplified at a time,compared with a conventional method, the cost is lower, the detection speed is higher, and meanwhile the success rate of detection is higher.