489 results about "Exon" patented technology

The invention relates to the technical field of plant genetic engineering, and discloses a haplotype identification marker of a rice flood-resistant gene ARF9 and application of the haplotype identification marker. The molecular marker is derived from an exon of an ARF9 gene, is located at the 22009082bp and 22010019bp of a No.4 chromosome of rice, and has a basic group of C. The SNP molecular marker is related to the waterflooding resistance of a male parent oryza longistaminate of perennial rice, and the waterflooding resistance of the male parent oryza longistaminate with a locus genotype of CC is obviously higher than that of rice with a locus genotype of non-AA type. Therefore, the invention provides the loop-mediated isothermal amplification primer group for rapidly identifying the ARF9 gene haploid molecular marker, and the loop-mediated isothermal amplification primer group is used for identifying the ARF9 gene haploid molecular marker so as to rapidly screen out waterflooding-resistant rice. The invention provides a group of primers, the flood-resistant rice can be quickly identified by adopting the primers to perform loop isothermal reaction, the detection cost is low, the detection speed is high, the flood-resistant germplasm can be quickly screened in the rice seedling stage by adopting the method provided by the invention, a time-consuming flooding test does not need to be performed in the later stage, and the breeding process is greatly accelerated.

The invention discloses a method, a system and equipment for detecting internal tandem repeat and a storage medium, and the key points of the technical scheme are as follows: obtaining a first reference sequence according to at least one target exon sequence corresponding to a protooncogene, and obtaining a second reference sequence according to at least one target intron sequence corresponding to the protooncogene; comparing the sequencing data of the to-be-detected sample to the second reference sequence to obtain a first comparison result, extracting an uncompared sequence from the sequencing data according to the first comparison result, and comparing the uncompared sequence to the first reference sequence to obtain a second comparison result; and determining a first detection result according to the second comparison result and a first reference sequence, and performing false positive filtering on the first detection result to obtain a second detection result. According to the invention, false positive can be reduced so as to ensure the accuracy and reliability of subsequent analysis.

The invention relates to a double-gene rare variation and disease relevance prediction model and an establishment method and application thereof, and belongs to the technical field of biological medicines.The establishment method of the double-gene rare variation and disease relevance prediction model comprises the following steps that S1, a sample library is screened; s2, performing quality control on whole exome sequencing data (WES); s3, performing phenotype screening; s4, performing grouping design; s5, carrying out PheWAS logistic regression analysis; s6, performing Firth logistic regression analysis and verification; and S7, carrying out double-gene feature analysis and double-gene pathogenicity relevance prediction. The method for analyzing the correlation between the rare double-gene variation and all disease phenotypes is designed for the first time, and a new method is provided for screening hereditary pathogenic factors of various diseases.

The invention discloses an Indel marker related to a corn kernel protein content character and application of the Indel marker, and belongs to the technical field of plant molecular genetics and crop molecular design breeding. The Indel marker is located on a gdh2 gene of a chromosome 10 of a corn genome, and insertion or deletion of a nucleotide sequence as shown in SEQ ID NO.5 occurs in an intron region between a second exon and a third exon. By applying the molecular marker-assisted selection technology, high-protein new germplasms S3317 (the crude protein content is 17.0%) and JS231148 (the crude protein content is 15.6%) have been successfully cultivated, the increase amplitudes of the high-protein new germplasms and JS231148 are both more than 15% compared with those of the conventional control, and the lysine content reaches 0.41% and reaches the national high-quality protein corn standard.

The invention provides a DGAT1 gene SNPs marker for detecting sheep milk production traits and application thereof.Nucleotide sequence variation sites of the DGAT1 gene are detected with the first-filial generation of Dongfudrisheng sheep male * Hu sheep female as an object, a general linear mixing effect model is constructed, the correlation between nucleotide sequence variation and sheep milk production traits is researched, and the DGAT1 gene SNPs marker for detecting the sheep milk production traits is obtained. And digging a molecular genetic marker for regulating and controlling the lactation character of the sheep. Three SNPs (single nucleotide polymorphisms) sites are found in total; wherein two variation sites in the first intron are named as c.191 + 411 Cgt, and the second intron is named as c.191 + 411 Cgt; t (SNP1) and c.192-440 Cgt, and c. 292-440 Cgt; t (SNP2) and a variation site in the 17th exon are named as c.1461 Cgt; t (SNP3). For the SNP1 site, the average daily milk yield of the CT genotype ewe is increased by 8.2% and 17.04% compared with that of the CC genotype ewe and that of the TT genotype ewe respectively; for the SNP3 site, the milk fat percentage of the CC genotype ewe is improved by 0.314% compared with that of the CT genotype ewe. A new molecular marker can be provided for selection and improvement of lactation traits of the milk sheep, and the molecular marker has important application prospects in sheep screening or breeding.

The invention belongs to the technical field of gene detection, and particularly relates to application of a CFTR gene mutation combination, an amplification reagent and a product in preparation of a CF risk assessment product. The CFTR gene mutation combination consists of an intron region mutation combination and an exon region mutation combination, based on the CFTR gene mutation combination, the invention further develops an amplification reagent and a CFTR gene mutation detection product, the amplification reagent and the CFTR gene mutation detection product comprise a primer combination for multiplex PCR amplification of the CFTR gene mutation combination, and the primer combination has the characteristics of high accuracy, high specificity and high sensitivity, can accurately detect related gene mutation, and provides a reliable basis for risk assessment and diagnosis of CF. When the amplification reagent and the CFTR gene mutation detection product are combined with sweat chlorine detection for use, the diagnosis rate of CF can be remarkably improved, and the amplification reagent has important application value in clinical diagnosis of CF and is expected to provide more powerful support for early diagnosis and treatment of CF patients.

The invention provides an SNP (Single Nucleotide Polymorphism) marker for targeted identification of systemic lupus erythematosus of children and application of the SNP marker. A new SNP site (RS25671007) is identified in a child systemic lupus erythematosus patient through whole exon sequencing, the site is located at the 734th pair of basic groups of a PABPC3 gene, and mutation from C to T exists at the site, or mutation from threonine to isoleucine exists in coded amino acid. In addition, in-vitro cell model experiments prove that the SNP causes significant up-regulation of the antibody type transformation function, and the SNP can be used as a reference marker for risk stratification or medication guidance of early targeted diagnosis, detection, gene evaluation and the like of systemic lupus erythematosus of children.

Aspects of the disclosure relate to recombinant gene editing complexes comprising a recombinant gene editing protein and guide RNA (gRNA) that specifically hybridizes to a region of a C90RF72 gene (e.g., a region flanking a G4C2 repeat or within a exonic region of the gene).

The invention provides a primer probe set for digital PCR detection of non-small cell lung cancer EGFR mutation and application thereof, and relates to the technical field of digital PCR detection, the primer probe set comprises a primer pair and a probe for specific detection of EGFR gene exon 19 deletion mutation (19del) and exon 21 L858R point mutation, and the 19del mutation probe comprises a plurality of sequences to cover common subtypes. The primer probe group can be combined with a microfluidic digital PCR platform for use, a sample is divided into tens of thousands of microdroplets for amplification, and FAM, HEX, ROX and CY5 fluorescence channel signals are detected to realize absolute quantification. The system provided by the invention has high sensitivity and excellent linearity and precision, is particularly suitable for detecting extremely-low-frequency EGFR mutation in body fluid such as plasma, and has important application value in individualized diagnosis and treatment, curative effect monitoring and drug resistance evaluation of non-small cell lung cancer.

This vector for nucleic acid introduction contains a nucleotide sequence that encodes a Cas protein, a nucleotide sequence that encodes a guide RNA, and a nucleotide sequence of a nucleic acid to be introduced. The guide RNA targets an intron adjacent to the 5' side, with reference to a sense strand, of an exon that includes a stop codon. Also provided is a nucleic acid introduction method comprising a step for introducing a nucleic acid to be introduced through non-homologous end joining by targeting an intron that is adjacent to the 5' side, with reference to a sense strand, of an exon that includes a stop codon.

The invention belongs to the field of biological medicine, and relates to a guide agRNA for regulating RNA splicing. The invention provides an application of a guide agRNA (Ribonucleic Acid) in preparation of a medicine for changing expression of a target protein by cells of a subject. The guide agRNA recruits ADAR protein to the 3 'splice site or pseudo 3' splice site to edit the A base of the 3 'splice site or pseudo 3' splice site, thereby splicing the whole exon located at the 3 'splice site or flanking the pseudo 3' splice site of the intron from the precursor mRNA, thereby changing the level of mRNA encoding the target protein, and altering the expression of the target protein in the cell. The target gene pre-mRNA selective splicing is regulated and controlled to influence the function or expression of the functional RNA and the target protein of the target gene, so that the purpose of treating diseases is achieved.

The application provides a rice heart-white gene WCG1 and application thereof, and particularly relates to positioning, cloning, function verification and application of a WCG1 gene for controlling a rice kernel heart-white phenotype. A C / A natural variation exists at a ninth exon 986 site of the WCG1 gene, the variation site makes the rice kernel present a heart-white phenotype, that is, a kernel middle heart-white and edge transparent phenotype, and the phenotype is an excellent phenotype of "sake brewing good suitable rice" specially selected in sake brewing.

The application discloses a KASP molecular marker of a Nicotiana benthamiana NbSAG101a gene and application thereof. The KASP molecular marker is an InDel insertion and deletion marker of the Nicotiana benthamiana NbSAG101a gene, is located at 256-262 bp of a second exon of the Nicotiana benthamiana NbSAG101a gene, and the nucleic acid sequence of the KASP molecular marker is shown as SEQ ID NO: 4. A wild-type allele of the Nicotiana benthamiana NbSAG101a gene is C, contains a 7 bp sequence shown as SEQ ID NO: 4, and a mutant-type allele is A, and the 7 bp sequence shown as SEQ ID NO: 4 is deleted. The KASP marker provided by the application can accurately and quickly distinguish wild-type homozygotes, heterozygotes and mutant-type homozygotes of the NbSAG101a gene, has high specificity, high accuracy, fast detection speed, low cost and high flux.

This invention discloses an efficient RNA data preprocessing method to address the problems of low processing efficiency and low information density in high-throughput sequencing data. Its core steps include: (1) introducing a parallel processing scheme for high-throughput sequence data, rapidly mapping RNA-seq data to a reference genome to generate a BAM file; (2) extracting base sequences and expression levels and storing them as compact PKL format files; (3) extracting all exon position information by parsing the genome annotation file; (4) combining multi-sample expression level data to screen for highly variable exons and constructing a high-information-density feature list based on the sample set; and (5) accurately extracting target sequences from the preprocessed file based on this list. Compared to traditional methods, this innovative approach achieves triple optimization: full-process parallel processing for accelerated computation, high-compression data storage, and adaptive feature selection. Processing speed is increased by 3-5 times, and data volume is reduced by more than 90%, making it suitable for high-throughput RNA-seq data analysis with large sample sizes.

The invention discloses construction and application of a Primrose syndrome non-human animal model, and belongs to the technical field of animal models and disease research. According to the invention, through a CRISPR / Cas9 technology, a codon CAC for coding 596th histidine in a No.14 exon of a mouse Zbtb20 gene is subjected to site-directed mutagenesis into CGC for coding arginine, and a hybrid mouse model for simulating pathogenic point mutation (p.H596R) of human Primrose syndrome is constructed. The model can stably reproduce key clinical phenotypes of the Primrose syndrome, including overgrowth after adult, serum IGF-1 rise, memory dysfunction and anxiety behaviors, and shows abnormal hippocampal neuronal development and synaptic transfer related pathways and the like. The invention provides an important experimental tool for deeply revealing the pathological mechanism of Primrose syndrome, developing drug screening and treatment intervention research and the like.

The invention provides a single-chain and double-protein-expressed circular RNA construct and a preparation method and application thereof, and the single-chain and double-protein-expressed circular RNA construct comprises 5'and 3 'introns used for realizing RNA self-splicing and cyclization as a first kind of introns; the 5'and 3 'homologous arms are used as pairing sites in RNA molecules; the exon 1 and the exon 2, which are connected after being spliced, can be spliced and connected together under the action of the first type of introns to form a complete and continuous open reading frame; an interval sequence for providing spatial and adjustment sequence structures; the IRES is used for starting translation; a glycine-serine linker as a flexible linker; a green fluorescent protein for gene expression monitoring; a luciferase for producing light by a catalytic chemical reaction. All the advantages of the double-IRES system for generating natural complete protein are reserved, and the core defect of low efficiency can be thoroughly overcome.

The invention provides a culter turunx2b gene knockout method and application of the culter turunx2b gene knockout method in creation of intermuscular thorn-free mutants, and belongs to the technical field of gene editing. The invention provides an efficient gene knockout method which comprises the following steps: selecting two targets on a second exon of a culter turunx2b gene as target sites, designing and synthesizing two gRNAs, mixing the gRNAs with Cas9mRNA, introducing the mixture into a fertilized egg of culter turunx2b in an I cell stage in a microinjection manner, hatching, and screening mutated F0-generation culter turunx2b individuals. By utilizing the method disclosed by the invention, the F0-generation knockout efficiency is up to 87.5% under the condition of ensuring that the fertilized eggs are hatched to survive. The method disclosed by the invention is simple and easy to implement and simple to operate, the culter alburnus gene mutation strain can be rapidly obtained by utilizing the gene mutation method, and the method has important scientific significance for developing economic fish gene function research, revealing genetic development law and creating new germplasm.

The invention relates to the technical field of molecular biology, in particular to a methylation marker for diagnosis of diabetic nephropathy. Specifically, the methylation marker comprises a P site of a CAT gene promoter region and / or an E site of a CAT gene exon 1 region. Based on a detection system combining methylation sensitive restriction enzyme with quantitative PCR, the kit has the advantages of simplicity and convenience in operation, good repeatability, accuracy in quantification and easiness in standardization, and is suitable for low-invasive samples such as peripheral blood. By effectively distinguishing pure type 2 diabetes patients from diabetic nephropathy combined patients, a reliable technical means is provided for risk stratification, early intervention and dynamic monitoring of diabetic people, and the method has good clinical application prospects and popularization value.

The invention relates to the technical field of molecular markers, in particular to an SNP (Single Nucleotide Polymorphism) molecular marker for predicting the daily age character of a pig for correcting 100 kg weight and application of the SNP molecular marker. The molecular marker is Ggt at the 53th site, the 157th site and the 723th site of the 12th chromosome of the Sscrofa11.1 version of a pig genome; according to the present invention, the CHD3 gene has the A single nucleotide polymorphism (SNP), and the site is located in the CHD3 gene exon region and belongs to the nonsynymous SNV; whole genome association analysis (GWAS) is carried out on a group containing 428 pigs, it is found that the site is extremely remarkably related to the growth speed required for correcting the day age of 100 kg of the weight of an individual (P = 1.27 * 10 <-10 >), and the individual carrying G allele shows higher breeding value, can serve as an important marker for molecular assisted breeding, is not limited by factors such as age and sex, and can be used as a marker for molecular assisted breeding. The method is suitable for breeding excellent boars, and can accelerate the formation of populations with excellent growth performance and improve the breeding efficiency of improved breeds.

The application discloses a kind of methods, systems, equipment and media for analyzing EML4-ALK fusion gene type, belong to the technical field of fusion gene detection.The method comprises obtaining RNA sequencing data of a sample to be tested;According to the primer combination coverage area, obtain the primer coverage area sequence file;The RNA sequencing data is compared with the primer coverage area sequence file for the second time;The number of sequencing data that can be aligned to the exon sequence of two genes at the same time is counted.Using the method and system of the application, unknown type of fusion gene can be accurately detected, which is more conducive to the development of downstream research on the related mechanism and function of fusion gene.

The application belongs to the field of molecular biology, and particularly relates to a specific marker of an exon region of a Oplegnathus punctatus hmcn2 gene, a primer pair, a detection method, a kit and application. The specific marker of the exon region of the Oplegnathus punctatus hmcn2 gene is a nucleotide sequence of non-insertion and insertion specific markers of the exon region of the Oplegnathus punctatus hmcn2 gene DNA fragments, and the sequence is shown in bases of SEQ ID NO:1 and SEQ ID NO:2. The specific marker is applied to identification of female and male genetic sex of the Oplegnathus punctatus. The application establishes a method for realizing rapid genetic sex identification by rapidly detecting an exon DNA insertion variation of the Oplegnathus punctatus hmcn2 gene, and the method has a wide application prospect in the fields of sex identification of the Oplegnathus punctatus, preparation of high male seedlings and family selection breeding.

Antisense oligonucleotides for modulating UNC13A splicing (e.g., inhibiting the inclusion of UNC13A hidden exons in mature UNC13A mRNA), compositions comprising such antisense oligonucleotides, and methods of use are described. Also disclosed are pharmaceutical compositions comprising one or more antisense oligonucleotides, and methods for treating UNC13A-related diseases or diseases associated with TDP-43 dysfunction in a subject by administering the antisense oligonucleotides to that subject.

The invention relates to a full-length sequence amplification primer, an amplification method and a three-generation sequencing method of an HLA-I antigen gene, and belongs to the field of gene detection. The HLA-I antigen gene is subjected to PCR (Polymerase Chain Reaction) amplification by using 8 pairs of characteristic amplification primers. And each gene can be subjected to high-yield specific amplification on a full-length sequence containing complete intron and exon regions only through one round of amplification and enrichment, and 7.8 Kb long-fragment genes can be captured once at most. The experimental deviation caused by simultaneous amplification of a single gene by using a plurality of pairs of segmented primers and the indirect error caused by gene splicing are effectively avoided. The method comprises the following steps: carrying out quality inspection and purification on amplicon PCR (Polymerase Chain Reaction) products, mixing samples, constructing an HLA-I type antigen gene library, carrying out accurate and complete sequencing on the full-length sequence of the HLA-I type antigen gene by adopting a three-generation PacBio Sequel II platform, and ensuring that the Hifi reads base accuracy can reach 99% or above. The reading of HLA-I type antigen gene full-length sequence variation information and the haplotype analysis at a high resolution level can be realized.

Double-stranded ribonucleic acid (dsRNAi) agents that target exon 1 of the huntingtin (HTT) gene are provided.SOLUTION: A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of huntingtin (HTT), the agent comprising a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a certain specific nucleotide sequence, and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from another certain specific nucleotide sequence, provided are dsRNA agents wherein one or more lipophilic moieties are conjugated to one or more internal positions on at least one of the sense or antisense strands.SELECTED DRAWING: None

The invention provides a method and a kit for detecting full-length mRNA of a human cell telomerase subunit TERT transcript, and belongs to the technical field of biology. The invention provides a method for detecting human cell telomerase subunit TERT transcript full-length mRNA, a specific reverse transcription primer TERT rt1 is adopted for reverse transcription to obtain cDNA, and if a sixth exon and a seventh exon of the telomerase subunit TERT can be detected in the cDNA, it is judged that a sample to be detected contains the human cell telomerase subunit TERT transcript full-length mRNA. The method disclosed by the invention can be used for distinguishing the full-length mRNA of TERT mRNA and other shear isomer mRNA, and a new technical means is provided for more conveniently, accurately and indirectly evaluating the activity of telomerase in cells.

The invention discloses a mutant gene OsNRAMP5 capable of reducing cadmium content in rice grains and application of the mutant gene OsNRAMP5. The gene has a genome nucleotide sequence as shown in SEQ ID No.1, and the 8875149th nucleotide on the No.7 chromosome of the wild type of fragrant rice, namely the 10th exon splicing site of the LOCOs07g15370 (OsNRAMP5) gene, is mutated from A to T, so that the gene is subjected to variable splicing in the transcription process. The invention also discloses a protein coded by the nucleotide sequence and used for regulating and controlling low accumulation of cadmium in fragrant rice grains, and the protein has an amino acid sequence as shown in SEQ ID No.3. The OsNRAMP5 gene for regulating and controlling the accumulation of the harmful heavy metal element cadmium is cloned in the fragrant rice by utilizing a forward genetics technology, and the mutant gene not only can improve the content of fragrant substances in the fragrant rice, but also can greatly reduce the content of cadmium in the rice, so that the quality safety of the fragrant rice is ensured.

The present application relates to the technical field of gene mutation prediction, and discloses a gene mutation prediction method generated by multi-mode annotation, and the specific process comprises the following steps: carrying out mutation type annotation on input single-base mutation position information to obtain mutation basic information containing mutation types, then using an ANNOVAR annotation tool, SpliceAI splicing effect prediction software and reference mutation information of a function effect database to carry out multi-dimensional feature annotation, using Bayesian PCA based on the obtained multi-dimensional feature mutation data set to fill in the annotation data, then using an automatic engineering feature list and a separated feature selection list to carry out feature combination and screening, and obtaining a gene mutation prediction score after gradient generation tree algorithm. The present application can be used for predicting all non-synonymous exon mutations, has good performance in classifying rare benign mutations, and can identify a small amount of mutations with high pathogenic probability from a large amount of candidate mutations.

This invention relates to a primer set and method for high-throughput assessment of drug resistance levels in rodent populations. The invention provides a set of universal degenerate primers for amplifying the Vkorc1 gene in multiple rodent species. The primer set includes primers for amplifying exon 1, exon 2, and exon 3 of the Vkorc1 gene, respectively. By designing universal degenerate primers and coupling them with next-generation sequencing technology, this invention can simultaneously amplify the Vkorc1 gene coding region of at least 15 rodent species across genera and families. This significantly improves the species universality and detection throughput of rodent drug resistance monitoring, reduces the cost of large-scale screening, overcomes the limitation of Sanger sequencing in accurately analyzing heterozygotes with insertions and deletions, and enhances the ability to discover new resistance mutations. It provides a standardized and low-cost technical solution for efficient assessment of rodent drug resistance.

The invention provides a gRNA, an AAV expression vector and a CRISPR-CasRx system for treating glaucoma, and particularly provides (a) a gene editing protein or an expression vector thereof, the gene editing protein comprises a VI type Cas protein; and (b) gRNA or an expression vector thereof, the gRNA is RNA for guiding the gene editing protein to be specifically combined with an aqueous humor outflow related gene and / or an aqueous humor generation related gene in an eye tissue, the aqueous humor outflow related gene comprises Rock1 and Rock2, and the aqueous humor generation related gene comprises Aqp1 and Adrb2. The gRNA designed aiming at exon regions of aqueous fluid outflow related genes (such as Rock1 and Rock2) and aqueous fluid generation related genes (such as Aqp1 and Adrb2) has the characteristics of relatively high editing efficiency and relatively low off-target rate, and the combination of the VI type Cas protein and the gRNA designed by the invention can effectively prevent and / or treat glaucoma or ocular hypertension.

The application discloses a molecular marker, a primer, a detection method and application related to pig muscle fiber, rib number and back fat thickness, and relates to the technical field of molecular markers. The molecular marker is located in a MYLPF gene, and a sequence of the molecular marker is shown as SEQ ID NO: 4. The sequence is located at a first exon of a 202 transcript of the MYLPF gene, and is used for detecting SNP site mutation. The application determines a cause mutation affecting the circumference, area of pig muscle fiber, rib number and average back fat thickness by using a MYLPF gene promoter constructed to replace a double luciferase reporter vector. A specific primer is designed on the mutation site of the promoter and amplification is performed. The site is detected for polymorphism by using a restriction endonuclease NlaIII. According to the result of the polymorphism detection, differences in the circumference, area of muscle fiber, rib number and average back fat thickness among pig individuals are distinguished, and a large population scale sampling and determination is not needed.