123 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.

Disclosed herein is a polynucleotide construct comprising one or more primary endonuclease recognition sequences upstream and downstream of a multiple gene editing site that comprises a plurality of secondary endonuclease recognition sequences. The primary endonuclease recognition sequences facilitate insertion of the multiple gene editing site into a host cell genome. The secondary endonuclease recognition sequences facilitate insertion of one or more exogenous donor genes into the host cell.

The present disclosure provides a polynucleotide comprising an expression cassette of a membrane protein linked to one or more additional expression cassettes for expressing one or more additional genes, the expression cassette of the membrane protein comprising a transmembrane domain and an extracellular domain, the extracellular domain comprising a recognition sequence, and one or more additional expression cassettes for expressing one or more additional genes, the one or more additional expression cassettes being linked to the one or more additional expression cassettes. The recognition sequence can be specifically recognized by the antibody of the recognition sequence. The invention also provides a vector and a cell containing the polynucleotide, and a method for transferring an exogenous gene into a cell and screening the cell by using the polynucleotide or the vector.

Herein is reported a method for preparing a recombinant host cell library expressing bispecific antibodies using targeted integration into a host cell comprising an exogenous nucleotide sequence integrated at a site within a locus of the genome of the host cell, wherein the exogenous nucleotide sequence comprises a first and a second recombination recognition sequence flanking at least one first selection marker, and a third recombination recognition sequence located between the first and the second, recombination recognition sequence, and all the recombination recognition sequences are different whereby the host cell is transfected with a library of first vectors each comprising two recombination recognition sequences matching the first and the third recombination recognition sequence on the integrated exogenous nucleotide sequence and flanking two exogenous nucleotide sequences and at least one second selection marker, and a library of second vectors each comprising two recombination recognition sequences matching the second and the third recombination recognition sequence on the integrated exogenous nucleotide sequence and flanking at least two further exogenous nucleotide sequences; wherein one of the four exogenous nucleotide sequences encode a first light chain, one a second light chain, one a first heavy chain and one a second heavy chain of the bispecific antibody.

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 invention relates to a method for simultaneously amplifying a plurality of target nucleic acid sequences in a sample, said method having the step of: (S300) amplifying the target nucleic acid sequences by means of an isothermal strand displacement amplification process using pairs of flanked primers, a superprimer (10), and at least one first nicking enzyme. One pair of flanked primers is used for each target nucleic acid sequence to be amplified, and the pair of flanked primers is specific to the associated target nucleic acid sequence, each pair of flanked primers having a flanked forward primer and a flanked reverse primer; each flanked forward primer has a target-specific portion at the 3' end thereof and a a non-specific flank portion at the 5' end thereof, the target-specific portion being complementary to a portion of the associated target nucleic acid sequence; each flanked reverse primer has a target-specific portion at the 3' end thereof and a non-specific flank portion at the 5' end thereof, said target-specific portion being complementary to a portion of the associated target nucleic acid sequence; and the superprimer (10) has a binding portion (12) at the 3' end thereof and a recognition portion (14) at the 5' end thereof, said recognition portion (14) having a recognition sequence for the first nicking enzyme.

The present disclosure provides conjugates comprising a targeting moiety, such as an antibody, Fab fragment or single chain variable fragment (ScFv), and a lipid nanoparticle (LNP) encapsulating a therapeutic agent (i.e., payload) wherein the targeting moiety, such as an antibody, Fab fragment or ScFv, is conjugated to the lipid nanoparticle via a linker, and wherein the linker comprises an enzyme recognition sequence and a click product formed by a click reaction between a first click handle on the targeting moiety such as an antibody, Fab fragment or ScFv and a second click handle on the LNP.

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 present invention addresses the problem of providing: a method for producing useful substances in animal cells, the method increasing the expression of a target gene; animal cells in which the expression of the target gene is increased; a method for producing such animal cells; and a kit for expressing the target gene in animal cells. The present invention provides a method for producing useful substances, the method comprising: introducing into animal cells (A) a nucleic acid containing (a) an exogenous target gene encoding the useful substance, (b) an exogenous constitutive promoter, and (c) a recognition sequence to which a binding substance can bind, and (B) a complex of three or more transcriptional activators and a binding substance, or a nucleic acid encoding the complex; and culturing the resulting animal cells.

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 present disclosure provides conjugates comprising a targeting moiety, such as an antibody, Fab fragment or single chain variable fragment (ScFv), and a lipid nanoparticle (LNP) encapsulating a therapeutic agent (i.e., payload) wherein the targeting moiety, such as an antibody, Fab fragment or ScFv, is conjugated to the lipid nanoparticle via a linker, and wherein the linker comprises an enzyme recognition sequence and a click product formed by a click reaction between a first click handle on the targeting moiety such as an antibody, Fab fragment or ScFv and a second click handle on the LNP.

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.

Disclosed herein are engineered nucleases that bind and cleave a recognition sequence within intron 1 of a transferrin gene, and methods of using such engineered nucleases to produce a genetically-modified eukaryotic cell comprising a modified transferrin gene. Further provided are pharmaceutical compositions and methods for treatment of a variety of conditions through expression of a polypeptide of interest encoded by an exogenous nucleic acid molecule inserted in intron 1 of a transferrin gene and expressed under the control of the endogenous transferrin promoter.

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.

Disclosed herein are engineered meganucleases engineered to recognize and cleave a recognition sequence present in human mitochondrial DNA (mtDNA). The disclosure further relates to the use of such engineered meganucleases in methods for producing genetically-modified eukaryotic cells, and to a population of genetically-modified eukaryotic cells, wherein mutant mtDNA is modified or degraded.

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.

The present disclosure relates generally to devices and methods for rapid and accurate diagnosis of single nucleotide polymorphisms. According to one or more embodiments, a system for identifying a single nucleotide polymorphism (SNP) comprises an oligonucleotide primer having a SNP recognition sequence, a fluorophore conjugated to the oligonucleotide primer at a conjugation site to form a fluorophore-conjugated oligonucleotide primer, a polymerase for extending the oligonucleotide primer to form a double-stranded deoxyribonucleic acid (DNA) amplicon upon binding to an oligonucleotide strand having a complementary sequence to the oligonucleotide primer, and wherein the fluorophore-conjugated oligonucleotide primer is the only primer in the system such that the fluorophore emits a fluorescent signal only when the fluorophore-conjugated oligonucleotide primer binds to a sequence specifically complementary to the SNP recognition sequence.

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 relates to the technical field of medical detection, in particular to a nanoprobe for simultaneously detecting and inhibiting VCAM-1mRNA expression as well as a preparation method and application of the nanoprobe. The nano particle takes a nano Au particle as a core, and p-mercaptocyanophenyl (4-MBN) serving as a Raman signal internal standard molecule is modified on the surface of the Au core; the nano particle takes a gold layer as a shell, and a VCAM-1mRNA targeting recognition sequence segment is fixed on the Au shell; and the tail end of the VCAM-1 mRNA targeted recognition sequence segment is modified with a Raman reporter molecule Cy5. The recognition sequence segment on the nanoprobe can be specifically combined with VCAM-1mRNA to form a rigid double strand, so that Cy5 is far away from the surface of the gold core-shell nanoparticle, the SERS signal strength of Cy5 is weakened, the position of 4-MBN located on a spacing layer is unchanged, and the SERS signal of the 4-MBN is also unchanged. The expression of the VCAM-1 protein can be inhibited by the accompanying mRNA consumption in the detection process, so that the synchronous monitoring and treatment intervention of the atherosclerosis disease closely associated with the expression of the VCAM-1 are realized.

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.

The invention discloses a preparation method of a biological chip space bar code sequence and a nucleic acid space omics detection method. The preparation method comprises the following steps: 1) connecting a silane group containing a silyl ether bond to a biological chip carrier; 2) establishing a plurality of parallel X-channel micro-channels on the biochip carrier, then performing DNA synthesis in each X-channel micro-channel by adopting a phosphoramidite synthesis method, and forming a first spatial position recognition DNA sequence on each detection point; and 3) establishing a plurality of parallel Y-channel micro-channels vertical to the X-channel on the biochip carrier, then performing DNA synthesis in each Y-channel micro-channel by adopting a phosphoramidite synthesis method, and forming a second spatial position recognition DNA sequence on the first spatial position recognition DNA sequence on each detection point.

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.