19 results about "Genetic design" patented technology

The invention provides a multi-stream fusion gene sequence prediction method based on Mama and a double-coding strategy, and aims to improve semantic understanding and modeling capability of DNA sequences, so as to improve accuracy and generalization performance of protein expression value prediction. The method comprises the following steps: synchronously processing a forward chain and a reverse complementary chain of a DNA sequence: carrying out word segmentation and vectorization on the reverse complementary chain by adopting BPE coding to capture a semantic fragment; and the forward chain is subjected to one-hot coding and is linearly mapped into dense representation. Deep features of the two features are extracted through Mama Block and then are spliced and fused, then LSTM is input to model context dependence, and finally an expression value is output through a full connection layer. According to the method, the problems that biological semantic fragments are difficult to capture by One-Hot coding and functional domains are likely to be split by BPE coding are effectively solved. On the basis of a Mama framework, a local structure and global semantics are effectively combined, and the method is good at capturing long-distance dependence while low calculation complexity is kept, and is suitable for tasks such as gene design and protein expression regulation and control.

The invention relates to a multi-target gene detection method based on a liquid phase chip, which comprises the following steps: S1, target genes: selecting O-type serum antigens to synthesize key genes; s2, probe design: synthesizing key genes aiming at seven serotype antigens, and designing a specific primer pair; s3, Luminex microsphere coding and coupling: using MagPlex microspheres with different fluorescence codes, and fixing a probe on the surface of the microspheres through a carboxyl-amino coupling method; s4, carrying out multiplex PCR amplification and hybridization detection; s5, a data analysis algorithm is adopted, and a threshold value is set, wherein a negative control MFI mean value + 3 * standard deviation is a Cuff value; and positive judgment: the MFI value of the microsphere corresponding to a certain serotype is greater than or equal to Cuff, and the signal-to-noise ratio (S / N) is greater than or equal to 5. By adopting the technical scheme, the method is based on a Luminex liquid phase chip technology, target gene sequences are amplified through multiple PCR (Polymerase Chain Reaction), and high-flux specific detection on seven serotypes of Escherichia coli is realized by combining microsphere coding probe hybridization and a biotin-streptavidin signal amplification system.

The invention discloses an sgRNA composition, a vector and a method for synchronous editing of pig SLA-DMA and SLA-DMB genes, and belongs to the field of gene editing. Editing targets capable of being specifically recognized by Cas9 endonuclease are designed for SLA-DMA and SLA-DMB genes of pigs, sgRNAs corresponding to the targets are connected in series with tRNA-sgRNA-gRNA scaffold as a minimum repetitive unit, and then a carrier capable of achieving synchronous expression of the four sgRNAs and targeting the SLA-DMA and SLA-DMB genes at the same time is constructed. When the vector is transfected into porcine cells, the SLA-DMA and SLA-DMB genes can be subjected to synchronous mutation, and an important tool is provided for researching a synergistic effect mechanism of the porcine SLA-DMA and SLA-DMB genes and pig gene editing breeding.

The invention belongs to the technical field of biology, and particularly relates to a construction method and application of a gobiocypris rarus gnaq knockout strain, a CRISPR / Cas9 gene editing technology is adopted, and double-target sgRNA (target 1 induces-8bp deletion and target 2 mediates + 32bp insertion) is designed for gobiocypris rarus gnaq genes. The sgRNA sequence is optimized through ZiFiT target spot design software, and the target specificity is ensured through NCBI BLAST comparison. The sgRNA and Cas9 mRNA which are transcribed in vitro are jointly microinjected into a gobiocypris rarus fertilized egg, and an F0 generation chimera is obtained. After the F1 generation heterozygote selfing, the homozygous mutants gnaq-8 and gnaq + 32 which conform to the Mendel genetic law are successfully screened in the F2 generation. According to the invention, gnaq knockout in gobiocypris rarus is realized for the first time, the blank of gobiocypris rarus G protein signal channel research is filled, and an important model is provided for subsequent fish related gene function research (such as antiviral immunity and tumor mechanism).

This invention relates to a 3'RACE library construction and sequencing method and its application based on third-generation sequencing technology. The method includes: S1, using primers containing anchor sequences and Oligo d(T) to reverse transcribe sample RNA to obtain cDNA, which is then used as a template for specific amplification to obtain the target gene; S2, designing specific primers containing third-generation sequencing adapters and barcodes, using the target gene as a template for amplification, and constructing a high-throughput third-generation sequencing library; S3, performing third-generation sequencing and analysis on the library to obtain the complete 3' end sequence information of the target transcript. This invention abandons traditional vector construction and Sanger sequencing methods, leveraging the high throughput and long read length characteristics of third-generation sequencing to ensure that Race experiments obtain a large amount of long 3' end sequence information in a short period, improving the overall efficiency and data accuracy of the experiment, and laying the foundation for refined mRNA research.

The application discloses a primer, a probe composition for nontuberculous mycobacteria typing and application thereof, and provides a PCR primer and a probe for rapid detection and typing of 13 common NTM in clinic based on genome big data for carrying out pan-genomics analysis, and the specific genes of the common NTM in clinic are screened by using a pan-genomics method, and then typing primers and probes are designed based on the genes, so that the problems of insufficient accuracy and specificity in NTM typing are solved; the various NTM specific sequences screened guarantee the accuracy and specificity; the high sensitivity of the PCR method and the specificity of the sequences guarantee that various interference substances in the clinical samples can be avoided, and the clinical samples can be directly analyzed; the PCR detection is low in cost and low in equipment requirement, and can be carried out in primary hospitals, thereby providing an efficient and economical solution for the precise diagnosis of nontuberculous mycobacteria, the control of disease prevalence and the large-scale disease screening.

The invention provides a quality control primer group and a quality control method for a human-derived sample, and belongs to the technical field of gene sequencing, and the quality control primer group comprises a human-derived DNA quality control primer designed based on conserved genes of GJB2MD, GJB3MD, IVSM and 12SD, a human-derived RNA quality control primer designed based on conserved and highly-expressed housekeeping genes of GAPDH, ACTB and PARP2, and a sex quality control primer designed based on genes of SRY142, DYS14134, YSR240 and YSR186 on a Y chromosome. According to the scheme provided by the invention, multi-dimensional and full-process monitoring on the human-derived sample can be realized, manual operation errors such as reagent failure, reagent addition error, sample confusion and cross contamination in the existing sequencing process are reduced, and the reliability of a detected result is effectively improved.

The application discloses a kind of for sheep MSTN And SOCS2 The sgRNA of gene cytosine base editing and application thereof, the sgRNA includes nucleotide such as any one or several of SEQ ID NO.1~4.This application is respectively designed sgRNA guide sequence for sheep MSTN And SOCS2 Gene, introduce stop codon in target site using CBE system, select sgRNA guide sequence and its combination from which can efficiently target sheep MSTN And SOCS2 Gene, carry out gene editing efficiency verification on cell, and successfully obtain MSTN And SOCS2 Double gene editing goat by prokaryotic embryo microinjection technology, and gene editing sheep shows muscle fiber thickening, and fast growth rate.This application establishes a set of efficient, precise double gene editing technology system, provides key breeding material and technical support for quickly cultivating muscle developed, fast growth meat sheep new variety.

The invention discloses a rapid detection method for vibrio corallinolyticus based on digital PCR, and relates to the technical field of biological detection.The method comprises the following steps that primers and probes are designed and synthesized according to the pyrH gene of the vibrio corallinolyticus; the method comprises the following steps: extracting total DNA of a sample to be detected as template DNA, and configuring a reaction system; and segmenting the prepared reaction system into micro-droplets by using a micro-droplet segmentation technology, then setting a reaction program to carry out PCR amplification, and calculating the content of the pyrH gene in the sample to be detected in combination with an end point fluorescence signal statistical result. According to the method, the pyrH gene is detected based on a digital PCR technology to reflect the pathogenic activity of pathogenic bacteria, the specificity and accuracy of vibrio corallinolyticus detection are remarkably improved, and an efficient, economical and accurate technical means is provided for comprehensive prevention and control of coral diseases.

This invention discloses a rapid multiplex nucleic acid detection method and application for reproductive tract pathogens based on RPA-CRISPR / Cas12a technology, belonging to the field of biodetection technology. The method uses HPV, UU, CT, and MG as detection targets. It designs and screens corresponding RPA primers and crRNAs targeting the specific target genes of each pathogen, optimizing the optimal primer pairs such as HPV-F2 / R2 and the optimal crRNAs such as HPV-crRNA2. Furthermore, it optimizes the CRISPR / Cas12a reaction system with a Cas12a to crRNA molar ratio of 0.9:1 and a final concentration of 500 nM for the fluorescent ssDNA reporter molecule. This integrates RPA isothermal amplification with CRISPR / Cas12a targeted detection, constructing a closed two-stage reaction system to achieve single-tube multiplex detection. This method can complete the test within 45 minutes. The detection limit for HPV / CT / MG is 10 copies / µL, and for UU it is 10² copies / µL. It has no specific cross-reactivity, and the clinical validation shows up to 100% consistency with qPCR. It does not require sophisticated instruments, and the endpoint detection method can be adapted to rapid on-site testing and can also be extended to other nucleic acid testing scenarios.

The invention relates to the technical field of anthrax detection / monitoring, and particularly discloses an anthrax wild strain and vaccine strain identification nucleic acid detection method. Comprising the following steps: S1, screening differential fragments of bacillus anthracis and designing primers and probes: designing primer and probe combinations for sap, pXO1 and pXO2 genes by referring to a whole genome sequence of the bacillus anthracis in a database; s2, gene cloning and standard substance preparation; s3, determining a reaction system of triple qPCR; and S4, carrying out qPCR (quantitative polymerase chain reaction) amplification detection. According to the fluorescent quantitative PCR detection method disclosed by the invention, the specific three genomes are screened and verified to cooperate as the identification marker, and the matched specific primer and probe sequences with high specificity and sensitivity are designed, so that the method can be used for efficiently identifying the bacillus anthracis wild strain and the vaccine strain; the method has the advantage of realizing efficient and specific synchronous amplification and detection of three target genes in the same reaction system.

The invention discloses a method for improving screening accuracy of specific primers of Shigella by using fluorescent loop-mediated isothermal amplification and a high-resolution melting curve, and belongs to the field of molecular biology. The method comprises the following steps: designing a primer group through an ipaH gene, extracting Shigella genome DNA, taking the Shigella genome DNA as a template, carrying out a fluorescent LAMP amplification reaction on the designed LAMP primer group, and carrying out HRM analysis on an amplification product; the positive amplification curve in the amplification curves is in a typical S shape, the negative control is not amplified, or the peak time of the amplification curve lags the positive peak time by more than 30 min is screened as a specific primer; in an HRM melting curve, a positive template is in a single sharp peak shape, and the positive Tm value is 90.0-91.0 DEG C; the negative control does not have a specific melting peak or has a wide and blunt non-specific peak, and the difference value between the negative control Tm and the positive Tm is greater than or equal to 2.0 DEG C. Through dual verification of fluorescent LAMP and HRM accurate analysis, the advantages and disadvantages of the primers are accurately evaluated, accurate and efficient screening of the optimal primer group is realized, and sensitive and specific primers are provided for subsequent LAMP detection tests.

The invention relates to a gene design and evolution path inference method based on a self-adaptive constrained depth model, which comprises the following steps: generating a random gene sequence set through a preset algorithm rule, converting the random gene sequence set into an evolutionary sequence containing evolutionary information, namely an evolutionary vector, and calculating mutation robustness; constructing a deep self-encoding network model, dynamically determining the optimal prototype number of the network through small-batch iterative training by adopting a prototype clustering analysis method, and performing model parameter optimization based on training data; inputting the evolutionary vector into an encoder module of an auto-encoder to obtain a low-dimensional representation of the evolutionary vector, namely an evolutionary space; establishing a deep regression prediction model, and accurately calculating the protein expression level corresponding to each regulatory gene sequence; establishing a low-dimensional tensor according to the obtained low-dimensional representation of the evolutionary vector and the corresponding expression value, performing Monte Carlo sampling in the low-dimensional tensor, mapping the protein expression value to the fitness, and generating a fitness contour map containing an evolution trajectory, namely obtaining a fitness terrain containing evolution information; and dyeing the low-dimensional tensor by using mutation robustness, and selecting a gene sequence with required properties on the graph. According to the technical scheme, the efficiency problem of existing gene sequence design is effectively solved, and the problem that the fitness terrain cannot represent evolution information of each genotype is effectively solved.

The application provides a multi-flow fusion gene sequence prediction method based on Mamba and a double coding strategy, aims to improve the semantic understanding and modeling ability of DNA sequences, and thus improves the accuracy and generalization performance of protein expression value prediction. The method synchronously processes the forward chain and the reverse complementary chain of the DNA sequence: the reverse complementary chain is segmented and vectorized by BPE coding to capture semantic fragments; the forward chain is encoded by one-hot and linearly mapped to a dense representation. After extracting deep features by Mamba Block, the two features are spliced and fused, and then input into LSTM to model the context dependence, and finally output the expression value through the full connection layer. The method effectively solves the problem that One-Hot coding is difficult to capture biological semantic fragments and BPE coding may split the functional domain. Based on the Mamba architecture, the local structure and global semantics are effectively combined, the long-distance dependence is captured while keeping low computational complexity, and the method is suitable for tasks such as gene design and protein expression regulation.

The invention discloses a method for detecting copy number of trophoblast residual quantity and application of the method. The method comprises the following steps: (1) inoculating trophoblasts containing a BCR-ALB mutant genome into a culture medium for amplification; (2) extracting and detecting the concentration and the purity of the BCR-ALB mutant genome; (3) designing a primer and a probe aiming at the BCR-ALB mutant genome and the reference gene; (4) constructing a real-time fluorescent quantitative PCR reaction system to carry out a real-time fluorescent quantitative PCR reaction; (5) cloning the BCR-ALB mutant genome to a vector to obtain a plasmid standard substance, carrying out gradient dilution after linearization, and drawing a standard curve; and (6) calculating the copy number of the trophoblast residual quantity in the sample to be detected according to the Ct value of the sample to be detected and the standard curve. The detection method can quantitatively detect the copy number of residual trophoblasts, is high in sensitivity, and can distinguish fragment interference signals.

The application relates to a gene design and evolutionary path inference method based on an adaptive band constraint deep model. First, a random gene sequence set is generated and converted into an evolvability vector containing evolutionary information, and the mutation robustness thereof is calculated. A deep auto-encoding network is constructed, model parameters are optimized through prototype clustering and small batch training, the evolvability vector is encoded into a low-dimensional representation, and an evolvability space is obtained. Then, a deep regression model is established to predict the protein expression level corresponding to the gene sequence. The low-dimensional vector and the expression value are combined into a low-dimensional tensor, the expression value is mapped into fitness through Monte Carlo sampling, and then an adaptive fitness topographic map showing the evolutionary track is generated. Finally, the tensor is stained according to the mutation robustness, and the gene sequence with the target property is screened. The scheme improves the gene design efficiency, and solves the problem that the fitness topography is difficult to represent the genotype evolutionary information.

The invention relates to the field of corn breeding, and particularly discloses a corn breeding process design system and an implementation method, the corn breeding process design system comprises a core germplasm resource module used for storing and managing screened and identified high-quality corn core germplasm resources, the core germplasm resources are subjected to gene mining based on extreme traits, and the core germplasm resources are subjected to genetic analysis; the method is obtained through a mixed sample analysis method, and has definite general combining ability (GCA) and special combining ability (SCA) evaluation. The core database module is used for storing genetic composition information of core germplasm resources, and the genetic composition information comprises high-throughput phenotype data, combining ability data and heritability data; according to the method, bad resources are eliminated from phenotypes through multi-environment accurate evaluation, SCA, GCA and heritability evaluation are carried out on selected excellent resources according to genetic design, and a genetic information base of core resources is constructed in the process, so that a breeder can accurately push a target improvement basis according to high GCA and clear SCA rules, and the breeding efficiency is improved. Powerful support is provided for subsequent breeding work.

The invention provides a specific primer and probe for detecting bovine tuberculosis and a kit composed of the specific primer and probe, and belongs to the technical field of biological detection. Specific primer pairs and probes are designed for cattle CXCL10, IFN-gamma and GAPDH genes, and quantitative detection of the transcription level of CXCL10 and IFN-gamma in cattle in-vitro whole blood is realized by using a qPCR technology, so that cattle tuberculosis is diagnosed. The method greatly shortens the detection time (incubation only needs 4 hours), simplifies the operation process, eliminates the system error in the sample adding process by using the GAPDH gene as the internal reference, and improves the accuracy of the detection result.

This invention discloses primers, kits, and methods for rapid sex identification of the peach fruit borer based on specific molecular markers. The primer pairs are based on male-specific genes of the peach fruit borer. CpunM The design incorporates the following primer sequences: 5'-AGGCTTTCGACAAGCTTCTCTA-3' and 5'-CCTGATTTGCGTGATAAGTGGT-3'. Specific PCR detection enables reliable sex identification of peach fruit borer larvae, pupae, and adults. This invention provides efficient and reliable technical support for implementing sex-specific control strategies for peach fruit borer, including sex ratio monitoring, mating behavior research, and sterile insect technology.