2105 results about "Nucleic acid sequence" patented technology

The present invention provides a method and a kit for detecting a target nucleic acid to be detected by using a melting curve. The method includes designing a first primer, a second primer and a detection probe for the target nucleic acid sequence to be detected, performing PCR amplification in a PCR amplification system containing the first primer, the second primer, the detection probe, the sample to be detected, a nicking enzyme and a DNA polymerase, generating a reporter primer that is complementary to the detection probe to form a double-stranded product, obtaining the melting curve of the double-stranded product, and the melting curve of the double-stranded product is the melting curve corresponding to the target nucleic acid to be detected. The method of the present invention is a non-target-dependent melting curve method, and the melting point (T m value) of each double-stranded body can be calculated in advance, solving the problems of melting curve peak shift and easy misjudgment caused by the easy mutation of the target nucleic acid sequence to be detected.

The present invention provides a method and a kit for detecting a target nucleic acid to be detected through a melting curve. The method includes designing a first primer, a second primer and a detection probe for the target nucleic acid sequence to be detected, performing PCR amplification in a PCR amplification system containing the first primer, the second primer, the detection probe, the sample to be detected, a restriction endonuclease and a DNA polymerase, generating a reporter primer that is complementary to and pairs with the detection probe to form a double-stranded product, obtaining the melting curve of the double-stranded product, and the melting curve of the double-stranded product is the melting curve corresponding to the target nucleic acid to be detected. The method of the present invention is a non-target-dependent melting curve method, and the melting point (T m value) of each double-stranded body can be calculated in advance, solving the problems of melting curve peak shift and easy misjudgment caused by easy mutation of the target nucleic acid sequence to be detected.

The invention belongs to the field of molecular biology, and particularly relates to a transgenic soybean event LP207-1 and a detection method thereof. The invention provides a nucleic acid sequence for detecting a transgenic soybean event LP207-1, the nucleic acid sequence is selected from one or more of sequences SEQ ID NO: 1-7 or complementary sequences thereof, and the nucleic acid sequence is derived from a plant, a seed or a cell of the transgenic soybean event LP207-1. A representative sample of a seed of the transgenic soybean event LP207-1 is preserved in the China Center for Type Culture Collection on May 23, 2025, and the preservation number is CCTCC NO: P202516. The transgenic soybean event LP207-1 has good resistance to lepidoptera pests and can tolerate agricultural herbicides containing glyphosate and glufosinate-ammonium, economic loss caused by the lepidoptera pests can be avoided, and the yield is not reduced; and the breeding efficiency is enhanced, and the molecular marker can be used for tracking a breeding population and transgenic insertion fragments in offspring of the breeding population. The detection method provided by the invention can quickly, accurately and stably identify the existence of the plant material derived from the transgenic soybean event LP207-1.

The invention provides a specific T cell receptor targeting KRAS G12V mutant epitope peptide (such as an amino acid sequence as shown in SEQ ID NO: 2) and anti-tumor application of the specific T cell receptor. The specific T cell receptor is composed of two peptide chains alpha and beta and can be specifically combined with a VVGAVGVGK-HLA-A * 11: 01 compound. The invention also provides an antigen binding fragment of the specific T cell receptor, a nucleic acid sequence for coding the T cell receptor, a vector containing the nucleic acid sequence, an engineered cell for expressing the T cell receptor, a composition containing the T cell receptor and application thereof. In addition, the invention also provides a method for screening the KRAS G12V specific T cell receptor. The specific T cell receptor and the antigen binding fragment thereof provided by the invention can be used as an immune effect activator to stimulate the immune response of a body, so that the effect of resisting diseases such as tumors and the like is achieved.

The invention relates to a molecular marker related to the content of 2-acetyl-3-methylpyrazine in pig muscle and application of the molecular marker, belonging to the technical field of determination or inspection methods of enzymes, nucleic acids or microorganisms. The technical problem to be solved by the invention is to provide a molecular marker related to the content of 2-acetyl-3-methylpyrazine in pig muscle. In order to solve the technical problem, the molecular marker provided by the invention is obtained through genome-wide association (GWAS) analysis, the nucleotide sequence of the molecular marker is as shown in SEQ ID NO: 1, and the molecular marker has an Agt at the 6328556 position of a chromosome 9 of a pig genome; a nucleotide single base of G is mutated, and the mutation obviously affects the content of 2-acetyl-3-methylpyrazine in pig muscles. By detecting the genotype of a to-be-detected pig at the SNP site, early selection of the content of 2-acetyl-3-methylpyrazine in pig muscles can be realized.

A technique related to codon optimization is provided. In a method for optimizing nucleic acid sequences expressing proteins in a host, protein subsequence-nucleic acid subsequence pairs are obtained from collected highly expressed protein sequences and their encoding nucleic acid sequences to form a training set. And training a neural machine translation model by using the training set. The neural machine translation model realizes translation from an amino acid sequence to a codon sequence. A protein sequence needing codon optimization is cut into protein subsequences. And translating a protein subsequence from an amino acid sequence to a codon sequence by using the trained neural machine translation model. The translated subsequences are superimposed to merge into a full-length codon sequence. In the process of superimposing to synthesize the codon sequence, according to the frequency or number of synonymous codons corresponding to each amino acid position of the protein sequence, the synonymous codon with the highest occurrence frequency or number is selected as the optimal codon at the position.

The disclosure provides systems and methods for droplet processing. For example, a method can include providing a first droplet and a second droplet. In some cases, the first droplet may have a first concentration of a reagent and the second droplet may have a second concentration of the reagent. The second droplet may comprise a bead or a biological particle. The method can also include subjecting the first droplet and the second droplet to conditions sufficient to transfer the reagent from the first droplet to the second droplet, thereby decreasing the first concentration in the first droplet and increasing the second concentration in the second droplet.

The invention belongs to the technical field of living cell imaging, and discloses a living cell nucleic acid imaging system and method based on CRISPR (clustered regularly interspaced short palindromic repeats) and a nano antibody and application, the system comprises: a modified dCas protein expression vector containing n antigen polypeptide tags; the modified sgRNA and / or crRNA expression vector comprises a specific guide sequence; the fluorescent nano-antibody expression vector contains m fluorescent nano-antibody coding sequences which are connected in an operable manner, and fluorescent protein serving as an internal tag is inserted between nano-antibody splitting sites, can be combined with a polypeptide tag carried by dCas protein and can stably emit light; and the fluorescent nano antibody in a free state can be self-degraded, so that the fluorescent nano antibody does not emit fluorescence. According to the invention, the single-copy nucleic acid sequence, especially the imaging marker of the single-copy nucleic acid sequence in a living cell, can be visualized with single-molecule resolution and high signal-to-noise ratio.

The present invention relates to CRISPR / Cas based biosensing materials, assays, and methods. Specifically, the present technology relates to CRISPR / Cas-based ultra-sensitive detection methods for nucleic acid assays using specific molecular constructs, including constructs referred to as nucleic acid mediators, comprising single-stranded and double-stranded nucleic acid sequences of a cyclic conformation, and palindromic oligonucleotides. The materials and methods according to the invention can also be used to enhance the sensitivity of existing bioassays.

The present application provides a keratin and a nucleic acid sequence encoding the same. The present application further provides a keratin gel dressing comprising the keratin of the present application, bFGF and a gel matrix.

Disclosed herein are chimpanzee adenoviral vectors that include neoantigen-encoding nucleic acid sequences derived from a tumor of a subject. Also disclosed are nucleotides, cells, and methods associated with the vectors including their use as vaccines.

The invention discloses application of a GsRZ1A gene in improving alkali resistance of plants, and belongs to the technical field of genetic engineering. The nucleotide sequence of the GsRZ1A gene is as shown in SEQ ID NO: 1, and the coded amino acid sequence of the GsRZ1A gene is as shown in SEQ ID NO: 2. A transgenic line is obtained by constructing an overexpression vector of the GsRZ1A gene and transforming soybeans. A transgenic line is subjected to alkali stress treatment, and a result shows that the overexpressed GsRZ1A soybean line shows a faster relative growth rate, the total leaf area, the leaf chlorophyll content and the relative water content are all higher than those of a control line of an empty vector, the total root length and the total root surface area of a root system are also remarkably larger than those of a control group, and the antioxidant enzyme activity can be improved; the accumulation of superoxide anions and malonaldehyde is reduced, and the accumulation of osmotic regulation substances such as soluble sugar is increased to enhance the tolerance of the soybeans to alkali stress, so that the method has an important application prospect in alkali-resistant plant breeding.

The invention relates to the technical field of nucleic acid detection, in particular to a method for regulating and controlling CRISPR-Cas protein activity through temperature and application of the method, a CRISPR reaction system contains Cas protein, guide RNA, a nucleic acid reporter probe and a buffer solution, and then the cis-cleavage activity or trans-cleavage activity of the Cas protein is activated or inactivated by regulating and controlling the temperature of the reaction system; the guide RNA includes a guide RNA targeting a target nucleic acid sequence that does not contain a PAM site. Multiple detection can be realized only by using the same type of Cas protein, the design of PAM-free sgRNA or crRNA facilitates the selection of sgRNA or crRNA, meanwhile, the activity of PAM-free sgRNA or crRNA can be regulated and controlled by utilizing temperature rise, and the Cas protein with lower reaction temperature can be inactivated, so that the Cas protein cannot cause interference to the detection of the second stage.

The invention discloses a phoebe zhennan PzGGPS12 gene and application of the phoebe zhennan PzGGPS12 gene to improvement of yield and drought tolerance of plant terpenoids, and belongs to the field of genetic engineering, a key enzyme gene PzGGPS12 for catalyzing biosynthesis of the terpenoids in phoebe zhennan wood is screened and analyzed, and a full-length CDS nucleic acid sequence of the PzGGPS12 is obtained. Transgenic experiments prove that overexpression of the gene improves the relative content of tobacco monoterpenes and triterpenes. Short-term drought experiments prove that overexpression of the gene can promote biosynthesis of sesquiterpenes, diterpenes and triterpenes so as to enhance drought tolerance of transgenic tobacco. Therefore, the gene can be introduced into a plant as a target gene, the yield of terpenoids in the plant is increased, and the drought tolerance of the plant is enhanced so as to improve the variety of the plant.

The invention discloses a primer probe group and a kit for detecting southern rust pathogenic bacteria, namely puccinia polypoda, based on MIRA-LFIA and application of the primer probe group and the kit. The primer probe group comprises a forward primer, a reverse primer and a probe combination, and the nucleotide sequence of the forward primer is as shown in SEQ ID NO.1; the nucleotide sequence of the reverse primer is as shown in SEQ ID NO. 2; the nucleic acid sequence of the probe primer is as shown in SEQ ID NO. 3. The MIRA-LFIA detection method of the pathogenic bacteria is established on the basis of the primers, and the detection method has good specificity and sensitivity, short detection time and low cost, realizes visualization of interpretation results by combining lateral flow immunochromatography color development, and is especially suitable for field real-time monitoring. The detection method provided by the invention lays a foundation for early diagnosis of puccinia multiflora and development of a monitoring technology of pathogenic bacteria, and provides guarantee for high yield of corn.

The invention provides a sequencing linker, a sequencing linker compound and a method for multiple nanopore sequencing of a target nucleic acid sequence. Wherein the sequencing linker comprises a first chain and a second chain, the first chain comprises a sequencing guide sequence, a helicase binding sequence, a limiting structure, a first complementary sequence and a first primer sequence from the 5'end to the 3 'end in sequence, the limiting structure prevents helicase from moving, and the first chain is provided with a 3' free end; the second chain comprises a second complementary sequence and a restraint sequence or a sequence complementary with the nucleic acid sequence of the restraint sequence from the 5'end to the 3 'end in sequence, and the restraint sequence comprises a nucleic acid sequence of which the tail end is connected with a hydrophobic molecule; the first complementary sequence and the second complementary sequence are reversely complementary, and the sequencing linker is formed by annealing the first strand and the second strand. The sequencing linker and the sequencing linker compound are simple in construction process and high in yield.

The invention provides an H1N1 broad-spectrum mRNA (messenger ribonucleic acid) vaccine taking HA (hemagglutinin) and NA (neuraminidase) protein tandem as a target and a preparation method of the H1N1 broad-spectrum mRNA vaccine, and relates to the technical field of vaccine preparation, the amino acid sequence of the mRNA vaccine is shown as SEQ ID NO.1, and the nucleotide sequence of the mRNA vaccine is shown as SEQ ID NO.2, namely, the mRNA vaccine is obtained by connecting GGGSGGSGGGSGGGGS with conserved amino acid sequences of hemagglutinin HA and neuraminidase NA. The defects in the prior art are overcome, and the broad spectrum and the protection effect of the vaccine are improved, so that the vaccine can better cope with the immune escape of the latest variant in the future.

The invention provides a design method of a protein variant structure based on molecular dynamics, and belongs to the technical field of molecular dynamics simulation. A protein-standard nucleic acid compound ground molecule system and a protein-nonstandard sequence compound ground molecule system are respectively constructed, and based on a molecular dynamics simulation technology, protein conformation stability, low-energy conformation states, main change processes, collaborative movement and key residues of the protein-standard sequence system and the protein-nonstandard sequence system are compared, so that protein conformation stability and low-energy conformation states of the protein-standard nucleic acid compound ground molecule system and the protein-nonstandard sequence compound ground molecule system are obtained. The molecular mechanism of protein-nucleic acid interaction is analyzed, the recognition behavior of the protein on the standard nucleic acid sequence and the activation process of the standard nucleic acid sequence on the corresponding function of the protein are understood from the molecular level, and finally the protein variant with more related functions is designed.

The present disclosure relates to T cells engineered to comprise a heterologous nucleic acid sequence encoding a dual mutant transforming growth factor beta 1 (dmTGFB1) under control of a promoter sequence and a modification, e.g., knockdown, of an endogenous nucleic acid sequence encoding a TGFBR2. In certain embodiments, the cells further comprise a modification, e.g., knockdown, of an endogenous nucleic acid sequence encoding a TNFA; a modification, e.g., knockdown, of an endogenous nucleic acid sequence encoding an IFNG; and insertion of sequence(s) encoding a regulatory T cell promoting molecule, and compositions and uses thereof.

The invention belongs to the technical field of molecular breeding of aquatic products, and particularly relates to an Indel molecular marker for identifying production places of finless eels and application thereof, which are particularly suitable for identifying finless eel strains in Jiangxi regions and non-Jiangxi regions. The Indel molecular marker for identifying the producing area of the ricefield eel is an Indel molecular marker B, and the Indel molecular marker B contains nucleotide sequences as shown in SEQ ID NO: 3 and SEQ ID NO: 4. The monopterus albus strains in Jiangxi regions and non-Jiangxi regions can be rapidly identified by utilizing the molecular marker, genetic identification can be rapidly realized, domestication breeding of wild germplasm resources is accelerated, and precise breeding is realized.

The invention belongs to the technical field of molecular breeding of aquatic products, particularly relates to an Indel molecular marker for identifying ricefield eel strains and application thereof, and is particularly suitable for identifying ricefield eel strains in Jiangxi regions and non-Jiangxi regions. The Indel molecular marker for identifying the ricefield eel strain is an Indel molecular marker A, and the Indel molecular marker A contains nucleotide sequences as shown in SEQ ID NO: 3 and SEQ ID NO: 4. The monopterus albus strains in Jiangxi regions and non-Jiangxi regions can be rapidly identified by utilizing the molecular marker, genetic identification can be rapidly realized, domestication breeding of wild germplasm resources is accelerated, and precise breeding is realized.

Methods, systems, and software for estimating circulating tumor fraction are provided. A first plurality of nucleic acid sequences for a plurality of loci in genomic DNA from a solid tumor sample is obtained. A second plurality of nucleic acid sequences for a plurality of cell-free DNA fragments obtained from a liquid biopsy sample from the same subject is obtained. One or more somatic mutations is identified in the first plurality of nucleic acid sequences. A variant allele frequency (VAF) is determined for each somatic mutation based on a frequency of the respective somatic mutation in the liquid biopsy sample and a frequency of the corresponding wild type allele in the liquid biopsy sample, thereby determining a set of VAFs. An estimate of the circulating tumor fraction for the test subject is determined based on the set of VAFs for the one or more somatic mutations.

Methods of labelling one or more subcellular components (e.g., an organelle and / or subcellular region) in vivo are provided. Methods of labelling a protein in vivo are provided. Methods of determining a nucleic acid sequence in situ are also provided.

The invention discloses an oil palm U6 promoter gene and application thereof, and belongs to the technical field of biology. The nucleotide sequence of the promoter gene is shown as SEQ ID NO.1. The oil palm RNA polymerase III type promoter gene, namely the oil palm endogenous U6 promoter gene EgU6, is obtained by cloning in an oil palm genome for the first time, and the promoter gene has high transcriptional activity and can drive downstream fluorescent protein mNeonGreen expression. The candidate oil palm endogenous U6 promoter gene can be provided for subsequently establishing a high-efficiency oil palm gene editing technology system based on a CRISPR / cas9 (Clustered Regularly Interspaced Short Palindromic Repeats / Cas9) system.

The invention discloses application of a peanut AhNF-YC9 gene in improving the oil content and salt tolerance of plants, and belongs to the technical field of gene engineering. The nucleotide sequence of the peanut AhNF-YC9 gene is as shown in SEQ ID NO: 1, and the coded amino acid sequence of the peanut AhNF-YC9 gene is as shown in SEQ ID NO: 2. The AhNF-YC9 gene is used for constructing a plant expression vector to transform peanuts, so that the AhNF-YC9 gene is excessively expressed in the peanuts, and the oil content of obtained transgenic peanut seeds can be increased by 4.0% at most compared with that of non-transgenic seeds; after stress treatment with 350mM NaCl, the salt tolerance of the transgenic peanut is obviously higher than that of a non-transgenic plant. Therefore, the peanut AhNF-YC9 gene has an important application prospect in improving the oil content and the salt tolerance of the plant.

The invention discloses keratinase Phyto-Keratinase and application of the keratinase Phyto-Keratinase. The keratinase Phyto-Keratinase comprises the following components: (1) protease which is coded by a nucleotide sequence as shown in SEQ ID NO.1; and / or (2) protease which is derived from bacillus subtilis, is coded by a nucleic acid sequence with more than 98% of identity with the SEQ ID NO.1 nucleotide sequence and has the characteristic of hydrolyzing keratin. According to the plant keratinase Phyto-Keratinase disclosed by the invention, the Phyto-Keratinase (plant keratinase) produced by utilizing microbial fermentation is a serine proteolytic enzyme, is also a biocompatible cutinase, and is obtained by fermenting saccharomyces cerevisiae. Phyto-Keratinase provides a nitrogen source for the growth of plants (beans), and contains various amino acids (functional peptides) and special proteolytic enzymes derived from the plants at the same time. The Phyto-keratinase can provide the effects of resisting inflammation, removing cutin and providing nutrition for skin regeneration at the same time, can relieve various inflammations by inhibiting PAR-2 receptors, and has potential value in the fields of cosmetics, beauty medicine and environment.

Systems and methods for determining a blood tumor mutational burden (bTMB) for a test subject are provided in which there is obtained, from a panel-enriched sequencing reaction, a plurality of nucleic acid sequences. The plurality of nucleic acid sequences comprises a corresponding sequence for each cell-free DNA fragment in a plurality of cell-free DNA fragments obtained from a liquid biopsy sample from the test subject. Each respective cell-free DNA fragment in the plurality of cell-free DNA fragments corresponds to a respective probe sequence in a plurality of probe sequences used to enrich cell-free DNA fragments in the liquid biopsy sample in the panel-enriched sequencing reaction. There is determined, using the panel-enriched sequencing reaction, that a circulating tumor fraction (ctFE) is above a threshold ctFE value. Responsive to this determination, the bTMB is calculated for the test subject from the panel-enriched sequencing reaction and reported.

Methods of labelling one or more subcellular components (e.g., an organelle and / or subcellular region) in vivo are provided. Methods of labelling a protein in vivo are provided. Methods of determining a nucleic acid sequence in situ are also provided.

The invention relates to a Kluyveromyces cerevisiae gene modified cell containing GDP-mannose dehydratase polypeptide, which can be applied to synthesis of human milk oligosaccharide, and belongs to the field of synthetic biology. According to the technical scheme, the gene modified cell comprises a recombinant nucleic acid sequence for coding a polypeptide with GDP-mannose dehydratase polypeptide (GDP-mannose dehydratase polypeptide) activity. The polypeptide capable of catalyzing the conversion of GDP-mannose into GDP-4-keto-6-deoxymannose comprises a polypeptide of which the amino acid sequence is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2 and / or the amino acid sequence of polypeptides M1-M13, wherein the amino acid sequence of the polypeptide is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to the amino acid sequence of the polypeptides M1-M13. The invention provides a kluyveromyces kluyveromyces genetically modified cell with fucosyllactose synthesis activity.