230 results about "Recognition sequence" patented technology

The invention provides an integrated electrochemical biosensor capable of detecting double miRNA with high sensitivity, and the integrated electrochemical biosensor can simultaneously detect double miRNA markers with high sensitivity through self-assembly of a Y-shaped capture probe, preparation of an Au / TiO2-FTO array electrode, catalytic hairpin amplification (CHA) and construction of a micro-droplet chip electrochemical biosensor. The advantages of equalization of double-target miRNA capture probe sites, excellent electrode performance, reliable signal amplification, low sample consumption and the like are integrated, the linear range of miRNA21 and miRNA155 detection is 10 fM to 100 nM, and the detection limits are 5.00 fM and 5.17 fM respectively. By detecting the miRNA21 and miRNA155 double markers, the accuracy of breast cancer diagnosis is up to 98%, and the recognition sequences at the two ends of the Y stent probe are modified, so that the sensor can be used for diagnosing other miRNA marker related diseases. The integrated micro-droplet chip electrochemical biosensor constructed by the invention has a remarkable clinical diagnosis value and a prominent practical application prospect.

The invention discloses a preparation method and application of a universal renewable immunoaffinity magnetic bead, and belongs to the technical field of chemical detection.The preparation method comprises the steps that firstly, an engineered bacteriophage H6G5-M13 is constructed, then a polypeptide module is coupled to the engineered bacteriophage H6G5-M13, the polypeptide module comprises a part A, a part B and a part C from the N end to the C end, the part A is a target object recognition sequence located at the N end, and the part C is a target object recognition sequence located at the C end; the part B is a selectively added connecting sequence, the part C is an LPXTG substrate sequence and is located at the C end, and X refers to any amino acid; finally, the engineered bacteriophage is directionally coupled to the Ni < 2 + >-IDA magnetic bead through the action of Ni-6His, and then the universal and novel immunoaffinity magnetic bead is formed. The universal and replaceable immunoaffinity magnetic bead prepared by the method can be used for preparing a chemical detection reagent, and can also be used for capturing, releasing and sensing detection of cells and extracellular vesicles.

Provided is a circular multimeric tandem RNA sense strand, comprising at least one sense strand sequence and at least one spacer sequence. The circular RNA is derived from an engineered parental DNA template containing all essential sequences, and sequentially comprises a first cyclization element, optionally at least one first restriction enzyme recognition sequence, at least one target sequence, optionally at least one second restriction enzyme recognition sequence, and a second cyclization element. The circular multimeric tandem RNA sense strand can bind and deliver a plurality of antisense strand RNA, increasing the binding of the sense strand and the antisense strand while utilizing the stability advantage of circular RNA.

The invention provides a three-generation sequencing read analysis method, application and device. The method comprises the steps that a read segment is obtained, the read segment is obtained by sequencing a sequencing library through a three-generation sequencing platform, and molecules of the sequencing library comprise a sequencing primer recognition sequence, a tag sequence and a UMI sequence; based on the sequencing primer recognition sequence in the read, determining an identity marker area of the read; comparing the identity marking area with at least one standard tag sequence, and determining the tag sequence of the read section in the identity marking area based on the comparison result; and determining the UMI sequence of the read based on the tag sequence in the identity marking area. According to the method, the characteristic of high insertion and deletion error rate in a three-generation full-length transcriptome sequencing result is fully considered, the utilization rate of three-generation sequencing data is improved based on a sequence alignment method, and the accuracy of transcript quantification is improved based on K-mer UMI sequence correction.

The invention provides annular multi-tandem PCSK9 siRNA, which comprises at least one positive-sense strand sequence and at least one spacer sequence. The sense strand of the cyclic siRNA is derived from an engineered parent DNA template containing all essential sequences, comprising in the following order a first cyclization element, optionally at least one first restriction enzyme recognition sequence, at least one target sequence, optionally at least one second restriction enzyme recognition sequence, and a second cyclization element. The annular multi-tandem RNA positive-sense strand can combine and deliver a plurality of antisense strand RNAs, and the combination of the positive-sense strand and the antisense strand is increased while the stability advantage of the annular RNA is utilized.

The present disclosure provides a method for analyzing a polyribonucleotide, wherein the polyribonucleotide is an mRNA molecule with a poly(A) tail, said method comprising: (a) obtaining a labeled polyribonucleotide by GI tailing; followed by (b) providing a second molecule comprising a first primer recognition sequence followed by a sequence of C nucleotide residues linked to a sequence of T nucleotide residues; followed by (c) obtaining a complex of said labeled polyribonucleotide and said second molecule; followed by (d) obtaining an extended second molecule by (d1) extending the 3′ end of the second molecule by synthesizing a sequence that is complementary to the labeled polyribonucleotide; followed by (d2) extending the 3′ end of the second molecule by adding at least 1 to 5 C nucleotide residues, followed by a second primer recognition sequence; said method providing information on entire polyribonucleotides, in particular mRNA isoforms and their respective poly(A) tails.

Provided herein are technologies for framing and evaluating biological sequence-based analysis tasks in a unified, natural-language-based, text in and text out format. Among other things, methods and systems of the present disclosure provide machine-learning technologies for combining biological sequence data, representing, for example, DNA, RNA, and protein sequences, with natural language, conversational style prompts that set out particular analysis tasks to be performed on the biological sequence data. This approach, for example, allows complex analysis tasks, including, but not limited to, identification of various sequence modifications, genes, and regulatory elements in DNA sequences, and quantification of properties such as degradation propensity of RNA and protein stability, to be input to a machine learning model in a uniform text-based format and for output to be generated in a same, unified, text-based format.

Provided is a circular concatemeric PCSK9 siRNA, comprising at least one sense strand sequence and at least one spacer sequence. The sense strand of the circular siRNA is derived from an engineered parental DNA template containing all essential sequences, which comprises, in the following order: a first cyclization element, optionally at least one first restriction enzyme recognition sequence, at least one target sequence, optionally at least one second restriction enzyme recognition sequence, and a second cyclization element. The sense strand of the circular concatemeric RNA can bind and deliver a plurality of antisense strand RNAs, enhancing the binding between the sense and antisense strands while leveraging the stability advantage of the circular RNA.

The invention discloses an information storage method based on DNA (deoxyribonucleic acid) coding, which comprises the following steps: converting digital information to be stored into a quaternary code, converting the quaternary code into a DNA sequence consisting of adenine A, cytosine C, guanine G and thymine T according to a preset mapping rule, and expressing an additional information dimension by using a chemical modification state of a nucleotide pair; inserting error detection and correction codes in the DNA sequence every a predetermined number of basic group positions, segmenting the encoded DNA sequence into a plurality of fragments with the length of 100-150bp, and adding a specific recognition sequence and an index marker at two ends of each fragment; a reversible thermosensitive response DNA nanostructure is used as a carrier, DNA fragments are selectively combined to the carrier, and hierarchical storage and rapid retrieval of information are realized through a temperature gradient control system; the DNA storage information density can be remarkably improved, the service life of DNA storage can be remarkably prolonged, and a novel technical path is provided for large-scale, long-term and safe molecular information storage.

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 invention discloses a method for site-specific replacement or insertion of DNA fragments in a plant genome, and belongs to the technical field of biology. The method disclosed by the invention comprises the following steps: editing a genome of a plant protoplast by adopting a CRISPR / Cas9 system and taking a double-stranded DNA repair template as a donor template to obtain an edited protoplast; and carrying out culture regeneration on the edited protoplast by utilizing a plant protoplast transformation method to obtain a site-directed mutagenesis plant in the genome so as to realize site-directed mutagenesis of the target plant genome. The method disclosed by the invention can be used for accurately replacing DNA in a plant genome at a fixed point. According to the invention, a recognition sequence of recombinase can be accurately inserted into a genome at a fixed point through homologous recombination, and then accurate fixed-point replacement of a gene large fragment is realized by virtue of a specific recombinase system.

The invention relates to the technical field of molecular biological detection, and discloses an isothermal enzyme-free cascade amplification system based on an HCR-DNAzyme technology and a method for detecting tetA by adopting the system. The system comprises an identification module, an amplification module, a feedback loop module and a detection module. According to the system and the method provided by the invention, HCR and DNAzyme technologies are integrated, a substrate is cut by DNAzyme to regenerate an initiation chain, a self-catalytic feedback loop is formed, signal exponential amplification is realized, the design of a hairpin structure (H1-H4) is optimized through NUPACK software, non-specific signal leakage is inhibited in combination with a blocking chain (B), and an identification sequence of an auxiliary hairpin (Hh) is replaced, so that the identification sequence of the auxiliary hairpin (Hh) is identified. Different ARGs detection (such as blaTEM-1 and aac3-II) can be rapidly adapted, and the repeated development cost is avoided.

The invention belongs to the technical field of biochips, and particularly relates to a reusable genetic typing chip and a detection method thereof, the reusable genetic typing chip provided by the invention is prepared from polystyrene microspheres, capture probes and a silicon substrate, and depends on competitive hybridization of DNA in an SNP recognition stage, and the detection result is more accurate. The recognition sequence of the capture probe is converted into the sequence of the target object, and the added detection probes are hairpin probes with a self-protection function, so that the generation of false positive signals is remarkably reduced, and the detection specificity is remarkably improved. And no enzyme participates in the whole process, so that the detection process is remarkably simplified, and the detection cost is reduced.

The invention discloses an application of ribonuclease SiRe0902 with high efficiency and thermal stability in specific recognition of a GG site. The amino acid sequence of the gene is as shown in SEQ ID NO.2, and the nucleotide sequence of the gene is as shown in SEQ ID NO.1. The ribonuclease SiRe0902 has very efficient and specific RNA (Ribonucleic Acid) hydrolysis activity, and does not have non-specific cutting on DNA (Deoxyribose Nucleic Acid) at all. In addition, the enzyme is derived from thermophilic archaea, is detected to have good thermal stability, and has very good RNA cleavage activity under the conditions of 37 DEG C and 65 DEG C. Through detection, the enzyme can specifically recognize a GG sequence for cutting, and also has high cutting activity for the GG sequence in a secondary structural region. Therefore, the ribonuclease SiRe0902 is novel ribonuclease with thermal stability, has extremely high RNA hydrolytic activity under normal-temperature and high-temperature conditions, and meets the requirements in the fields of clinic, production, molecular cloning and the like.

A single nucleotide polymorphism typing method based on a CRISPR / Cas12a logic gate realizes typing output of single nucleotide polymorphism through logic gate input of fluorescence signals of a CRISPR / Cas12a reaction system, namely ''existence'' and ''no'', ''strong'' and ''weak'', and is characterized in that the novel single nucleotide polymorphism typing method comprises two steps of target fragment amplification and CRISPR / Cas12a detection; wherein the target fragment amplification is characterized in that a PAM recognition sequence TTTN of CRISPR / Cas12a is introduced to a-15 site to a-11 site of a target site through the design of an amplification primer, and an amplification product can be detected by the CRISPR / Cas12a; according to the CRISPR / Cas12a detection, two crRNA1 and crRNA2 which can distinguish wild type and mutant type amplification products are used as guide chains of a CRISPR / Cas12a reaction system, a novel functionalized single-stranded DNA report probe is designed, and single nucleotide polymorphism typing is achieved through logic gates of'existence 'and'absence' and'strong 'and'weak' of fluorescence signals. The method solves the problem that a traditional CRISPR / Cas12a reaction system cannot realize single nucleotide polymorphism typing, can be used for scientific research analysis and clinical detection, and has a wide market prospect.

The present disclosure encompasses engineered nucleases which recognize and cleave a recognition sequence within a Hepatitis B virus (HBV) genome. The engineered meganucleases described herein can exhibit improved characteristics, such as enhanced specificity and / or efficiency of indel formation, when compared to previously described HBV meganucleases. Further, the disclosure encompasses pharmaceutical compositions comprising engineered meganuclease proteins, nucleic acids encoding engineered meganucleases, and the use of such compositions for treating HBV infections or diseases associated with HBV infections.

The embodiment of the invention provides a library construction method, a universal joint and a kit, relates to the technical field of synthetic biology, and is used for constructing a mixed library in which multiple gene segments are directionally connected. The library construction method comprises the following steps: providing a plurality of first initial gene segments; introducing cutting sequences to two ends of each first initial gene segment to obtain a plurality of second initial gene segments; introducing universal joints to two ends of each second initial gene segment to obtain a plurality of target gene segments; wherein the universal joint comprises an identification sequence; performing cutting and connection reaction on the plurality of target gene segments to obtain a mixed library; wherein the plurality of target gene segments are connected according to a set sequence. The library construction method is used for constructing the DNA mixed library.

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 invention encompasses engineered meganucleases which recognize and cleave a recognition sequence within an open reading frame (ORF) of the genome of at least two genotypes of the Hepatitis B virus (HBV). The present invention also encompasses methods of using such engineered meganucleases in a pharmaceutical composition and in methods for treating or reducing the symptoms of a HBV infection, or treating hepatocellular carcinoma (HCC). Further, the invention encompasses pharmaceutical compositions comprising engineered meganuclease proteins, nucleic acids encoding engineered meganucleases, and the use of such compositions for treating HBV infections or HCC.

Provided is a nucleic acid structure which enhances an antigen-specific immune response. The nucleic acid structure comprises a polynucleotide encoding a SNARE protein, a polynucleotide encoding a proprotein convertase recognition sequence, and a polynucleotide encoding an antigen, wherein the polynucleotide encoding a SNARE protein and the polynucleotide encoding an antigen are linked via the polynucleotide encoding a proprotein convertase recognition sequence.

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 provides application of a ring finger protein coding gene OsRF in regulation and control of rice grain shape and grain weight, the gene OsRF for coding the ring finger protein is separated and cloned from rice Nipponbare, and the gene is targeted by rice small RNA OsmiR530. In order to prevent the OsmiR530 from being cut and improve the expression level of the OsmiR530, artificial synonymous mutation is carried out on the OsRF recognized sequence by the OsmiR530, a mutant OsRF fragment (mOsRF) of which the recognition sequence is subjected to synonymous mutation is created by utilizing overlapping PCR (Polymerase Chain Reaction), an overexpression vector of the mOsRF is constructed and is overexpressed in rice, and the grain width and the grain weight of transgenic rice are obviously increased. Therefore, the overexpression of the mOsRF in the rice is of great significance for obtaining round-grain rice and increasing the rice yield, and a new resource and a new research method are provided for rice grain shape improvement breeding.

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 present invention discloses an information storage method based on DNA coding. The method comprises the following steps: converting digital information to be stored into a quaternary code, and converting the quaternary code into a DNA sequence consisting of adenine A, cytosine C, guanine G, and thymine T according to a preset mapping rule, and using the chemical modification state of the nucleotide pairs to represent an additional information dimension; inserting error detection and correction codes at every predetermined number of base positions in the DNA sequence, dividing the encoded DNA sequence into multiple fragments of 100-150 bp in length, and adding specific recognition sequences and index markers at both ends of each fragment; utilizing a reversibly thermoresponsive DNA nanostructure as a carrier, selectively binding the DNA fragments to the carrier, and realizing hierarchical storage and rapid retrieval of information through a temperature gradient control system; the present invention can significantly improve the information density and lifespan of DNA storage, and provides a new technical path for large-scale, long-term, and secure molecular information storage.

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.