61 results about "Recognition sequence" patented technology

The invention discloses a method for improving the solubility and thermal stability of sweet protein, and belongs to the technical field of biosynthesis, the method comprises the following steps: constructing a gene tandem recombinant plasmid containing a monellin x-3C-sfGFP-3C-monellin y expression cassette, x is greater than or equal to 1, y is greater than or equal to 1, 5 is greater than or equal to x + y is greater than or equal to 3, a 3C protease recognition sequence is also inserted between adjacent copies of monellin, and each monellin is connected with a purification tag; the gene tandem recombinant plasmid is transformed into escherichia coli and inducible expression is carried out, thalli are split and purified to obtain fusion protein, 3C protease is used for enzyme digestion, and the target protein is obtained after re-purification. The total protein yield and the solubility proportion are remarkably improved by adopting a series construction mode of plasmids, meanwhile, the stability of monellin is effectively improved, the purification step is simpler and more convenient, and the method is suitable for industrial production.

The present disclosure encompasses a lipid nanoparticle (LNP) comprising a polypeptide comprising a nucleic acid sequence encoding an engineered meganuclease that binds and cleaves a recognition sequence within a Hepatitis B virus (HBV) genome. Further, the disclosure encompasses pharmaceutical compositions comprising the LNPs, and the use of such compositions for inactivating a pol gene of an HBV genome or an HBV genome fragment in a cell and treating HBV infections or diseases associated with HBV infections.

The present application discloses an antiviral fusion protein. Specifically, the present application provides an antiviral fusion protein, which comprises a cell death inducing domain (lethal domain), a domain inhibiting the activity of the lethal domain (inhibitory domain) and a protease recognition sequence capable of removing or destroying the function of the inhibitory domain; the fusion protein has the activity of inducing cell death after being cut by a specific viral protease, thereby killing the cells infected by the virus in a targeted manner. The present application also discloses a programmable method for inducing the death of virus-infected cells, which comprises transferring the above-mentioned fusion protein or nucleic acid into cells so that the cells die before replication and assembly after being infected by the virus, thereby achieving timely and effective elimination of the virus.

The application is suitable for the biotechnology field, and provides a multiplex amplification method and system and application, wherein the multiplex amplification method comprises the following steps: linear amplification of a target template by using an upstream primer mixture designed for multiple target regions to obtain a first product by purification; the 5' end of the upstream primer contains a universal primer sequence, and the 3' end contains a specific target recognition sequence; linear amplification of the first product by using a downstream primer mixture designed for multiple target regions to obtain a second product by purification; the 5' end of the downstream primer contains a universal primer sequence, and the 3' end contains a specific target recognition sequence; and unified exponential amplification of the second product by using a universal primer. The application innovatively adopts a specific multi-target molecular marker combined with a universal primer amplification strategy to establish a brand-new multi-target DNA fragment amplification technology system, and high uniformity, high specificity and high-throughput multi-target amplification can be realized.

The application relates to the technical field of molecular markers, and discloses a litchi dwarfing trait related gene SNP molecular marker, a primer thereof and application. LcGAMYB23 The litchi dwarfing trait related gene is , the SNP molecular marker is derived from a polymorphic site of the gene, has the advantages of high polymorphism, good genetic stability and simplicity and efficiency, and the site recognition sequence is CAAAATTTTC[T / G]GTCTGCTGGA. The application further discloses a primer pair for detecting the SNP molecular marker, the sequence of an upstream primer is shown as SEQ ID NO. 1, and the sequence of a downstream primer is shown as SEQ ID NO. 2. The SNP molecular marker and the primer pair can be used for quickly identifying the dwarfing trait of litchi germplasm resources, can significantly accelerate the breeding rate of litchi dwarfing varieties, and promote the creation, utilization and genetic improvement of dwarfing resources.

The purpose of the present invention is to provide a polypeptide capable of binding to protein L. Provided is a protein L-binding polypeptide comprising amino acid sequence 1, which is L1 sequence (such as the amino acid sequence represented by SEQ ID NO: 1) - antigen recognition sequence 1 - L2 sequence (such as the amino acid sequence represented by SEQ ID NO: 2) - antigen recognition sequence 2 - L3 sequence (such as the amino acid sequence represented by SEQ ID NO: 3) - antigen recognition sequence 3 - L4 sequence (such as the amino acid sequence represented by SEQ ID NO: 4).

The invention is applicable to the technical field of biology, and provides a multiple amplification method and system and application wherein the multiple amplification method comprises the following steps: performing linear amplification and purification on a target template by using an upstream primer mixture designed for a plurality of target regions to obtain a first product; the 5'end of the upstream primer contains a universal primer sequence, and the 3 'end of the upstream primer contains a specific target recognition sequence; performing linear amplification on the first product by using a downstream primer mixture designed for a plurality of target areas, and purifying to obtain a second product; the 5'end of the downstream primer contains a universal primer sequence, and the 3 'end of the downstream primer contains a specific target recognition sequence; and performing uniform exponential amplification on the second product by using a universal primer. According to the invention, a brand new multi-target DNA fragment amplification technology system is established by innovatively adopting a specific multi-target molecular marker and combining a universal primer amplification strategy, and high-uniformity, high-specificity and high-throughput multi-target amplification can be realized.

This solution discloses a new method for identifying RNA pseudouridine sites. This method proposes to use a variety of feature representation techniques to extract sequence features, and then uses the SVM-RFE method for feature selection to compress the feature space and optimize the feature subset. The best feature set after feature selection is input into the kernel method KeMRF based on polynomial random forest to identify pseudouridine sites in the sequence. As a newly proposed classification method, compared with the traditional random forest, KeMRF not only optimizes the discriminant criterion for node splitting, but also combines with an easy-to-interpret kernel method, making the classification performance more superior. This method reduces the training time of the model, improves the classification performance of the model, and further enhances the accuracy of identifying pseudouridine sites.

The invention relates to an adaptor, a composition, a kit and a method for nucleic acid amplification detection. The adaptor, the composition, the kit and the method can be used for quantitative detection of nucleic acid. Specifically, the adaptor comprises a universal sequence (2s) and a binding capture sequence (1s) from the 5'end to the 3 'end; wherein (1) the binding capture sequence (1s) is at least partially complementary with a sequence on a target molecule, (2) the adaptor further comprises a nucleic acid extension blocking modification located at the 3'end of the binding capture sequence (1s), and (3) the binding capture sequence (1s) has (including) 1-2 enzyme cleavage recognition sequences or modifications close to the 3 'end. The invention also provides a composition or a kit, a detection system and a method for nucleic acid amplification detection. The product and the method provided by the invention can realize nucleic acid detection with good specificity and high sensitivity.

This invention discloses a method and kit for detecting RNA using a three-stage amplification melting curve based on isothermal self-circulation. The method includes designing a first primer, a second primer, and a circular amplification probe targeting the target nucleic acid sequence. The first primer consists of an RNA polymerase promoter sequence and a target nucleic acid-specific binding sequence, sequentially from the 5' end to the 3' end. The circular amplification probe is a circular DNA segment whose sequence consists of three parts: the first part completely contains a continuous sequence composed of the tag sequence and the nicking enzyme-specific recognition sequence from the second primer; the second part is a self-cleaving DNase sequence. In the technical solution used in this invention, a cascaded nucleic acid amplification system is constructed, achieving exponential signal amplification of the target nucleic acid through the synergistic effect of rolling circle amplification and RNA polymerase-mediated transcriptional amplification.

The application relates to a method and device for identifying a modification site in an RNA sequence, computer equipment and a medium. The method comprises the following steps: obtaining sequence features, structure features of the RNA sequence to be identified, and a pre-trained modification site identification model; inputting the structure features into a structure convolution layer for convolution to obtain convolution structure features; in each attention encoding unit: inputting the sequence features into a query weight matrix, a key weight matrix and a value weight matrix respectively for linear transformation, thereby obtaining query features, key features and value features; fusing the query features, the key features and the convolution structure features to obtain first fusion features; fusing the first fusion features and the value features to obtain second fusion features; splicing the second fusion features output by all the attention encoding units to obtain spliced features; and performing feature adjustment on the spliced features based on a feature adjustment module to obtain an identification result. The method can improve the accuracy of the identification result.

There is provided a polynucleotide-modifying enzyme with a functional nuclease domain and a display domain. The functional nuclease domain comprises a nuclease catalytic pocket. The display domain comprises a peptidic recognition sequence of from 3 to 20 amino acids in length, in a loop, an alpha helix or an extension off the end of the alpha helix that is positioned on an external surface of the polynucleotide-modifying enzyme. The peptidic recognition sequence recognizes a target cell receptor of a target cell to allow cell internalization of the polynucleotide-modifying enzyme in said target cell.

Provided herein are systems and methods for transposase-mediated insertion of an adapter construct into a target nucleic acid. A composition can include a nucleic acid molecule, comprising a plurality of barcode sequences and a plurality of capture sequences. The composition can further include a plurality of transpososomes coupled to the nucleic acid molecule, wherein each transpososome comprises an active transposase and a plurality of polynucleotides. The plurality of polynucleotides can include: (A) a pair of transposon end recognition sequences that are bound to the active transposase and (B) a polynucleotide comprising a binding sequence that is hybridized to a capture sequence of the plurality of capture sequences of the nucleic acid molecule.

The invention discloses a preparation method of a proteoglycan protein conjugate carrying a specified connection site. The method comprises the following steps: carrying out recombinant expression on a recombinant tetanus toxin heavy chain C fragment rTTHc containing a Sortase A 7M ligase recognition sequence, introducing O, O '-(ethane-1, 2-diyl) bis (hydroxylamine) as a nucleophilic reagent into the C tail end of the rTTHc under the action of ligase Sortase A 7M, and carrying out specific condensation reaction on the C tail end of the rTTHc as a unique chemical reaction site and polysaccharide carrying an aldehyde group functional group to obtain the tetanus toxin heavy chain C fragment rTTHc. Therefore, polysaccharide protein conjugates with definite polysaccharide and protein connection sites are produced. According to the invention, the problem that the key antigen epitope of the carrier protein and the heterogeneity of the product are possibly damaged by the traditional random modification strategy is solved, the antigen epitope of the carrier protein is prevented from being damaged to a great extent, and a reliable technical scheme is provided for preparing the polysaccharide conjugate vaccine with high quality and high immunogenicity.

The invention belongs to the technical field of biomedical detection, and particularly relates to a DNA fiber bar code probe for multiple miRNA detection and application of the DNA fiber bar code probe. In order to overcome the defects that existing miRNA detection is low in sensitivity, small in flux, serious in cross interference and the like, the probe comprises a DNA fiber and a fluorescence signal probe, and the DNA fiber is formed by self-assembly of DNA tile monomers; a recognition sequence complementary with target miRNA extends from the 3'end of the DNA tile monomer, a fluorescent signal probe with a hairpin structure is matched, a fluorescent signal is released when the target miRNA exists, the signal intensity is in direct proportion to the miRNA concentration, different target miRNAs trigger the hairpin signal probe with the corresponding fluorescent color, a decodable bar code is formed through fluorescent color combination, multiple miRNA detection is achieved, and the miRNA detection efficiency is improved. Further, the tumor cell types are accurately distinguished.

Disclosed are recombinant meganucleases engineered to bind and cleave a recognition sequence present in a mutant RHO P23H allele. The invention further relates to the use of such recombinant meganucleases in a method for treating retinitis pigmentosa, wherein the mutant RHO P23H allele is preferentially targeted, cleaved, and inactivated.

The invention provides a method for detecting Ggt in gene mutation. The invention relates to a lock type probe for detecting A mutation, which comprises a recognition sequence complementary with a target p53 R175H and a sequence recognized by CRISPR / Cas12a, and is modified with 8-oxoguanine. The invention also provides a Ggt based on the combination of the Fpg enzyme and CRISPR / Cas12a (clustered regularly interspaced short palindromic repeats / CRISPR / Cas12a). The universal type detection kit for A mutation comprises the lock-type probe, a fluorescence report probe, a primer, crRNA, a T4 DNA ligase, an Fpg enzyme, a phi29 DNA polymerase and a Cas12a protein. The invention also provides a Ggt; the invention discloses an A mutation detection method and application. According to the invention, the Ggt based on the combination of the Fpg enzyme and CRISPR / Cas12a is successfully constructed; according to the universal detection method for the A mutation, a 8-oxoG modified lock-type probe is used for recognizing a mutation site, and rolling circle amplification and CRISPR / Cas12a cascade signal amplification system are combined, so that the core bottlenecks of insufficient universality caused by restriction enzyme dependence and strict dependence of Cas protein on a target PAM site in the existing mutation detection technology are solved in a breakthrough manner, and the purpose of detecting all Ggt, Ggt, Ggt, Ggt, Ggt, Ggt, Ggt, Ggt, Ggt, Ggt and Ggt mutation is achieved. And broad-spectrum and accurate detection of point A mutation is realized.

This disclosure covers engineered macronucleases that bind to and cleave recognition sequences within the dystrophin gene. This disclosure also covers methods for preparing genetically modified cells using such engineered macronucleases. Furthermore, this disclosure covers pharmaceutical compositions comprising engineered macronuclease proteins or polynucleotides encoding engineered macronucleases of this disclosure, and the use of such compositions for modifying the dystrophin gene in a subject or for treating Duchenne muscular dystrophy.

The invention discloses an editing system for accurate knock-in of a large-fragment gene and application of the editing system, the editing system can express dCas9-FokI fusion protein and EBNA1 protein, and comprises an oriP element, a region for inserting a double sgRNA coding sequence and a donor template sequence for knock-in of the large-fragment gene. According to the editing system, cut FokI nuclease is generated in a dimerization form, a recognition sequence of sgRNA is not damaged, dCas9-FokI nuclease and a donor template are continuously expressed by mediation of an EBNA1 / oriP attachment, a target sequence in which accurate large-fragment gene knock-in does not occur is continuously cut, DSB frequency is generated as many as possible, and meanwhile, the target sequence in which accurate large-fragment gene knock-in does not occur is continuously cut. Further, HDR is generated as much as possible, and the purpose of improving knock-in of large-fragment genes is achieved. By means of the editing system, large-fragment gene knock-in can be efficiently carried out, and a feasible method and a powerful gene editing tool are provided for gene function research.

This invention belongs to the field of genetic engineering technology, specifically relating to a recombinant type I collagen, its preparation method, and its applications. Its amino acid sequence is shown in SEQ ID NO.1. The recombinant type I collagen of this invention is based on a short amino acid sequence from natural type I collagen, with the integrin recognition sequence GFPGER added as a repeating unit, repeated multiple times, with a repetition count of 6. The recombinant type I collagen exhibits extremely good hydrophilicity and stability, high adhesion, and does not cause immune rejection when applied to the human body. It can be used in the preparation of tissue engineering materials such as bone implants and bone repair scaffolds.

Provided is a polynucleotide comprising an expression cassette of a membrane protein and one or more additional expression cassettes for expressing one or more additional genes. The expression cassette of the membrane protein is connected to the one or more additional expression cassettes. The membrane protein comprises a transmembrane domain and an extracellular domain. The extracellular domain comprises a recognition sequence. The recognition sequence can be specifically recognized by an antibody of the recognition sequence. Further provided are a vector comprising the polynucleotide and a cell, and a method for transferring an exogenous gene into the cell by using the polynucleotide or the vector and screening the cell.

The present invention encompasses engineered meganucleases that bind and cleave a recognition sequence within the first exon of the human T cell receptor (TCR) alpha constant region gene. The engineered meganucleases can exhibit at least one optimized characteristic, such as increased specificity or efficiency of cleavage, when compared to previous generations of meganucleases. The present invention also encompasses methods of using such engineered nucleases to make genetically-modified cells, and the use of such cells in a pharmaceutical composition and in methods for treating diseases, such as cancer.

The present invention provides a genome editing system for Corynebacterium glutamicum, comprising: a nucleic acid protein complex capable of specifically cleaving DNA in Corynebacterium glutamicum, the nucleic acid protein complex comprising endonuclease and a guide RNA targeting a target sequence; the endonuclease comprises an amino acid sequence as shown in SEQ ID NO: 1 or an amino acid sequence having at least 80% sequence identity with the amino acid sequence as shown in SEQ ID NO: 1; the guide RNA comprises (1) a nucleic acid sequence as shown in SEQ ID NO: 2 or a nucleic acid sequence having at least 80% sequence identity with the SEQ ID NO: 2, and (2) a recognition sequence complementary or identical to a target sequence, and the length of the recognition sequence is 5-50 nt. The system can realize genome editing in corynebacterium glutamicum, and the editing efficiency is better than that of Cas9 and Cpf1.

Herein is reported a novel variant adeno-associated virus (AAV) capsid protein comprising a heterologous peptide or polypeptide covalently inserted in the GH-loop of the capsid protein relative to a corresponding parental AAV capsid protein, wherein the heterologous peptide or polypeptide comprises a recognition sequence of a transglutaminase and the amino acids residues at positions 452-455 or 453-459 of VP1 of AAV-2 (SEQ ID NO: 65) or at the corresponding positions in the capsid protein of another AAV serotype are replaced by the heterologous peptide or polypeptide or inserted at amino acid position 456.

The invention relates to the technical field of nucleic acid drugs and gene therapy, in particular to a preparation method and application of endothelial cell specific modRNA. The system comprises a number 1 modRNA and a number 2 modRNA, the number 1 modRNA encodes a target protein, a 5 '-untranslated region of the number 1 modRNA comprises an L7Ae protein binding element k-turn, the number 2 modRNA encodes an L7Ae protein, a 3'-untranslated region of the number 2 modRNA comprises at least one miR126 recognition sequence, and the number 1 modRNA and the number 2 modRNA are used for being delivered into an endothelial cell; the system has no genome integration risk, the expression is regulated and controlled by miR126 in real time, and the system is suitable for vascular regeneration of targeted endothelial cells, precise gene therapy of ischemic heart disease and the like.

The invention belongs to the technical field of biology, and particularly relates to a novel fusion protein and application thereof in preparation of an ARHGAP9 inhibitor. The novel recombinant fusion protein comprises a core recognition sequence, a linker, a hinge region and a mutated immunoglobulin heavy chain constant region fragment. It is proved for the first time that the core recognition sequence-human RAC2-ECD structural domain of the novel recombinant fusion protein can be combined with ARHGAP9, and then the novel recombinant fusion protein can be prepared into a potential ARHGAP9 inhibitor. Finally, the invention also proves that the novel recombinant fusion protein has a positive effect in treating cancers as an ARHGAP9 inhibitor.

The invention discloses a method for efficiently enriching anti-blue ear and porcine reproductive capacity edited pig donor cells. The method comprises the following steps: introducing CRISPR-Cas9 plasmids targeting a BMP15 gene, CRISPR-Cas9 plasmids targeting a CD163 gene and alternative reporter plasmids edited by double genes into the pig donor cells; culturing the porcine donor cells to enable gene editing and reporter gene expression to occur; detecting and sorting cells with positive report gene expression so as to obtain enriched double-gene editing donor cells; the alternative reporter plasmid for double-gene editing comprises the following elements which are connected in sequence: a recognition sequence of a BMP15 gene editing target, a transcription terminator sequence, a recognition sequence of a CD163 gene editing target and a fluorescent reporter gene sequence. According to the invention, the alternative reporter plasmid for BMP15 and CD163 double-gene editing is constructed, and the editing condition of target sites of the BMP15 and CD163 genes of a cell genome can be converted into a detectable red fluorescence phenotype, so that the enrichment efficiency of anti-blue ear high-fecundity edited pig donor cells is improved, and the double-gene editing efficiency of the cells is effectively improved.

The present invention provides methods for generating a population of captured target nucleic acids and compositions for use in the methods. An exemplary method can include contacting a plurality of oligonucleotides individually linked to a solid support with a sample comprising a target nucleic acid having a 3'end and a 5 'end, where the target nucleic acid is annealed to some free 3' ends of the oligonucleotides, and where there is an excess of oligonucleotides such that at least some of the oligonucleotides still have free 3 'ends; then extending the 3'end of the target nucleic acid with a polymerase using an oligonucleotide as a template to form a double-stranded portion comprising a restriction enzyme recognition sequence; then cleaving the restriction enzyme recognition sequence with a restriction enzyme to form a released target nucleic acid having a single-stranded 5'end and a double-stranded 3 'end, and leaving an oligonucleotide not cleaved from the solid support and still having a free 3' end; the solid support and the uncleaved oligonucleotide are then optionally separated from the released target nucleic acid to form a target nucleic acid solution.

The present invention refers to methods for selectively recognizing a base pair in a DNA sequence by a polypeptide, to modified polypeptides which specifically recognize one or more base pairs in a DNA sequence and, to DNA which is modified so that it can be specifically recognized by a polypeptide and to uses of the polypeptide and DNA in specific DNA targeting as well as to methods of modulating expression of target genes in a cell.