77 results about "Functional validation" patented technology

The invention belongs to the technical field of plant genetic engineering, and discloses application of an OsSRO1c gene in controlling high-temperature resistance of rice. The OsSRO1c gene capable of improving the drought tolerance of rice is obtained through separation, cloning and functional verification, the nucleotide sequence of the OsSRO1c gene is as shown in SEQ ID NO.1, and the sequence of protein coded by the gene is as shown in SEQ ID NO.2. The gene OsSRO1c for controlling rice high-temperature response is cloned, CRISPR mutant phenotype identification is carried out on candidate genes, high-temperature stress phenotype identification in a seedling stage and an adult-plant stage shows that when a gene segment is deleted, the high-temperature resistance of rice is reduced, and when the expression quantity of the gene is increased, the high-temperature resistance of rice is improved, and the function and the application way of the gene are proved.

The invention discloses an EsSTB1 gene capable of increasing the number of branches of a plant, reducing the plant height and delaying flowering. The EsSTB1 gene has the functions of increasing the number of branches, reducing the plant height and delaying flowering, and a scientific basis and a new material are provided for cooperative regulation and control of tillering formation and reproductive growth of siberian wildrye; besides, the functional verification method of the EsSTB1 gene provided by the invention performs functional verification by applying transgenic arabidopsis thaliana starting from an internal regulation mechanism of tillering regulation of siberian wildrye, has obvious research depth and breadth and technical effects, and can obtain a plant strain with an intentional phenotype. Through verification, the siberian wildrye EsSTB1 gene disclosed by the invention can be used for remarkably increasing the number of branches of a plant, reducing the plant height and delaying flowering. The method is suitable for popularization and application in the biotechnology field.

The present application provides an isolation, cloning, functional verification of a restorer gene OsRf19 for cytoplasmic male sterility in rice and its use in improving rice. The present application specifically provides a nucleotide sequence of the gene OsRf19 and an encoded amino acid sequence thereof, an expression vector comprising the nucleotide sequence of the gene OsRf19, a method of creating a restorer line of rice using the gene OsRf19, and a method of restoring the fertility of a cytoplasmic male sterility line of rice and producing rice hybrid seeds, and related uses.

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 invention discloses a Chinese cabbage clubroot-resistant gene BrRLP31 and application of the Chinese cabbage clubroot-resistant gene BrRLP31. The nucleotide sequence of the Chinese cabbage clubroot-resistant gene BrRLP31 is as shown in SEQ ID No. 1. Functional verification results show that after the expression of the endogenous BrRLP31 of the Chinese cabbage is inhibited by utilizing a VIGS technology, the resistance of the plant to clubroot is obviously reduced, and the disease index and the root swelling severity of the plant are obviously higher than those of a control material. On the other hand, the transgenic flowering cabbage plant over-expressed by the BrRLP31 is constructed by an agrobacterium-mediated method, and disease resistance identification proves that the heterologous over-expression of the BrRLP31 obviously improves the resistance of the plant to plasmodiophora brassicae. The results show that the BrRLP31 gene plays a key positive regulation role in the clubroot resistance process of the Chinese cabbage, the disease resistance of cruciferae crops and other horticultural plants is improved by utilizing the gene and the homologous sequence thereof, and the gene and the homologous sequence thereof have important application value.

The application discloses a kind of BRCA1 / 2 The application discloses a function verification method of genetic unknown meaning variation. By introducing only one target variation in each experiment, combined with optimized gRNA and template design, the method ensures that the target variation has high initial abundance in the edited cell population, thereby significantly reducing the background noise of second-generation sequencing detection and phenotype analysis, improving the sensitivity and reliability of variation function signal detection. The method provides a more flexible, accurate and easy-to-operate function verification scheme for BRCA1 / 2 The method provides a more flexible, accurate and easy-to-operate function verification scheme for interpretation of unknown meaning variation, and is expected to solve the bottleneck of difficulty in obtaining and interpreting function data of unknown meaning variation.

The application belongs to the technical field of biology and specifically relates to a casein phosphopeptide for promoting calcium, iron and zinc mineral absorption and improving bone health. The application obtains the casein phosphopeptide containing high-activity peptide segments by optimizing the process, and takes six mineral elements of calcium, magnesium, copper, iron, zinc and manganese as research objects. Firstly, the mineral deficiency rat model is used to evaluate the mineral utilization of the CPP. The consumption of 7mg / kg.d increases the contents of Ca, Mg, Cu, Fe, Zn and Mn in femur by 7.12%, 3.56%, 20.76%, 15.84%, 9.94% and 67.12% respectively. Secondly, the molecular docking technology is used to screen the peptide segments with calcium absorption activity from the CPP and perform function verification. The application aims to reveal the effect of the CPP in the mineral absorption and utilization from the cell and animal levels, and provide a theoretical basis for the functional development.

The invention discloses application of a Homer 1 gene in regulating and controlling the water-holding capacity character of a pig, and belongs to the technical field of biology. Candidate genes related to the water-holding capacity are analyzed and screened through transcriptomics; and identifying proteins and metabolic markers related to the water-holding capacity in the longissimus dorsi aging process by utilizing metabonomics and proteomics technologies. And carrying out cellular level function verification on the screened key gene Homer 1 through multi-omics integration analysis. The result shows that the Homer 1 gene is negatively correlated with the water-holding capacity, so that a theoretical basis is provided for understanding a regulation mechanism of the water-holding capacity of the pig skeletal muscle; meanwhile, candidate gene resources and theoretical support are provided for breeding of high-quality and high-efficiency new pig varieties.

This disclosure provides a method and related products for genome sequence generation and model training. During the training phase, pre-trained multimodal data, including sequence data, functional annotation data, omics data, and species evolution information, is acquired. A fused representation is obtained through heterogeneous coding and multimodal fusion, and input into the sequence generation model for feature calculation, outputting the predicted base probability distribution for each base position to be generated. The model is optimized by combining a multi-task training mode of autoregressive generation and intermediate padding, and a weighted loss function based on biological annotation. During the inference phase, sequences are generated progressively based on multimodal generation context information. Candidate bases are obtained through sampling and evaluated using biological constraints, omics validation, structural validation, and functional validation. Resampling is performed when conditions are not met until preset conditions are satisfied. This method can improve the biological rationality and application value of the generated sequences.

The invention discloses a rape cold-resistant gene and application thereof, and belongs to the technical field of gene engineering. Four rape specific lone genes (BnaOGs) and 2859 brassica specific genes (BSGs) are identified through comparative genomics and multi-database artificial verification by combining UniProt-KB, Nrdb and the like, and the characteristics of short sequence, high GC content, no transcription factor and the like are found. Aiming at a BSG gene BnaA03G0334200ZS, a CDS sequence of the BSG gene BnaA03G0334200ZS is cloned, a pEGOEP35S-H overexpression vector containing a 35S strong promoter and a hygromycin selection marker is constructed, and a positive plant is obtained through agrobacterium-mediated transformation of brassica napus Westar. Functional verification shows that overexpression of the gene remarkably enhances the cold resistance of the rape, plant damage is slight at low temperature, the relative conductivity and MDA content are reduced, and the activities of SOD, POD and CAT are improved. The invention provides a new target and technical support for cultivation of cold-resistant rape varieties, and has a wide application prospect.

This invention discloses a method for wheat grain quality detection and functional gene analysis based on hyperspectral imaging technology, belonging to the field of agricultural testing technology. The method includes: extracting hyperspectral indices; measuring wheat grain quality indicators as manual measurements; assigning weight coefficients to each quality indicator to digitize the wheat quality phenotype; extracting characteristic wavelengths to establish predictive models for each indicator and wheat quality values; performing GWAS on hyperspectral indices and chemical indicators as phenotypic traits with genotypes to screen SNP loci, conduct co-location analysis, obtain candidate genes, and perform functional verification. This invention, through hyperspectral technology combined with chemometric modeling, is suitable for real-time monitoring during wheat quality breeding and processing; by combining spectral phenotype with GWAS gene mining, it overcomes the limitations of traditional quality detection that only serves phenotypic screening, forming a closed loop of "non-destructive detection - gene localization," providing targeted genes for wheat quality improvement.

The invention belongs to the technical field of gene engineering, and particularly relates to application of CmIAA27 in regulation and control of lateral branch development of chrysanthemum. According to the invention, phenotypic analysis is carried out on a CmCMT2-RNAi transgenic 'Shenma' plant obtained at an earlier stage, and lateral branches of the CmCMT2-RNAi transgenic 'Shenma' are increased in a vegetative growth period. RNA-seq is carried out on the gene CmIAA27, it is found that the expression quantity of the auxin inhibitory signal key gene CmIAA27 is remarkably increased, McrBC and BSP experiments show that the CHH type methylation level on a CmIAA27 promoter in a transgenic line is remarkably lower than that of WT, and the expression of the plant lateral branch development key inhibitory gene CmBRC1 in the transgenic line is remarkably reduced. Y1H, LUC and EMSA experiments show that the CmCMT2 can be directly combined with a CmIAA27 promoter and regulate and control the CHH type methylation level in the region so as to influence the expression of the CmIAA27. Through transcription analysis of CmCMT2-RNAi transgenic Shenma and WT, a key factor CmIAA27 for downstream regulation and control of chrysanthemum lateral branch development is obtained, functional verification is carried out, and excellent gene reserve and a new strategy are provided for regulation and control of chrysanthemum lateral branches.

The application discloses a pichia pastoris bioengineering bacterium for relieving degradation of recombinant collagen, a preparation method and application thereof. Through bioinformatics analysis, gene knockout and function verification screening, three protease genes capable of relieving degradation of collagen in a pichia pastoris host are obtained, namely, chr1-4_0611, chr1-4_0362 and chr2-2_0380. Knocking out the protease chr1-4_0611 can make the total protein yield accumulation increase to 3.5 times of that of a control bacterium, and the proportion of a target band is greater than 50%, which can provide good theoretical and technical guidance for effective expression of other recombinant proteins in the pichia pastoris host.

The invention relates to an application of a corn ZmXTH27 gene in improving the cold resistance of plants. The ZmXTH27 gene encodes a protein of a xyloglucan endotransglycosidase / hydrolase (XTH) family, the gene expression level of the ZmXTH27 gene is remarkably increased under low-temperature stress, and a mutant zmxth27 of the gene shows a low-temperature sensitive phenotype. The ZmXTH27 gene is transformed into arabidopsis thaliana and is subjected to functional verification, and compared with wild type arabidopsis thaliana, the tolerance of an arabidopsis thaliana plant over-expressed with the ZmXTH27 gene to cold stress is remarkably enhanced. According to the invention, more theoretical bases are provided for corn cold resistance research, and gene resources and biotechnological means are provided for subsequent cultivation and germplasm resource innovation of new cold-resistant corn strains.

The invention discloses a non-specific peroxygenase mutant and an application thereof. According to the method, Collariella virescens UPO of which the protein structure is analyzed is rationally designed by utilizing a multiple computer-aided strategy, and through functional verification, a UPO mutant library which has rich enantioselectivity to various substrates and is improved in activity is successfully obtained, so that a solid foundation is laid for researching and developing a green and efficient chiral compound production method. Compared with the prior art, the non-specific peroxygenase mutant provided by the invention shows high catalytic activity in selective oxidation of carbon-hydrogen bonds of various substrates, the selectivity is remarkably improved or overturned, the obtained reaction product is high in optical purity, the reaction efficiency is improved, the separation process is simplified, the cost is reduced, and the non-specific peroxygenase mutant is suitable for industrial production. Good industrial application prospects are realized.

The application discloses a non-specific peroxygenase mutant and application thereof. The application uses a multiple computer-aided strategy to rationally design Collariella virescens UPO of a resolved protein structure, and successfully obtains a UPO mutant library with rich enantioselectivity to various substrates and improved activity through function verification, thereby laying a solid foundation for researching and developing a green and efficient chiral compound production method. Compared with the prior art, the non-specific peroxygenase mutant provided by the application has high catalytic activity in the selective oxidation of carbon-hydrogen bonds of various substrates, and the selectivity is significantly improved or reversed, the obtained reaction product has high optical purity, the reaction efficiency is improved, the separation process is simplified, the cost is reduced, and the non-specific peroxygenase mutant has a good industrial application prospect.

The invention belongs to the technical field of plant genetic engineering and molecular breeding, and particularly discloses a key negative regulation gene GhHSFC1 for regulating verticillium wilt resistance of cotton and application of the key negative regulation gene GhHSFC1. The gene is positioned on a chromosome A05 of upland cotton of a genetic standard line TM-1, the coding sequence (CDS) of the gene is shown as SEQ ID NO. 1, and the sequence of a protein coded by the gene is shown as SEQ ID NO. 2. Through bioinformatics analysis and in combination with a core virus-induced gene silencing (VIGS) technology, functional verification is carried out, and it is confirmed for the first time that the GhHSFC1 gene is a negative regulatory factor of cotton verticillium wilt resistance. According to the gene resource and the application thereof provided by the invention, a novel target spot subjected to functional verification and a precise and efficient improvement strategy are provided for molecular breeding of cotton verticillium wilt resistance, and the gene resource has important research value and application prospect.

This invention discloses a method for screening and regulating environmentally adaptive minor genes during wheat propagation, comprising the following steps: Step S1: Define the target environment and adaptability phenotype, extract key environmental covariates of the target ecological zone, and construct a digital environmental profile; screen physiological and ecological traits corresponding to environmental stress and measure them using high-throughput phenotyping technology; Step S2: Construct genetic materials and design experiments, using a genetic material system composed of a core germplasm bank, a multi-parental design population, and chromosome segment substitution lines, and setting up multi-environment gradient experiments and precise stress control experiments; Step S3: Integrate data analysis and candidate gene mining, integrate environmental covariates, and screen genetic loci that change synergistically with environmental signals; determine gene information strongly correlated with adaptability phenotypes through weighted gene co-expression network analysis; Step S4: Functional verification and mechanism analysis to confirm the environment-specific function of the genes.

The invention belongs to the fields of biotechnology, protein engineering and plant protection science, and particularly relates to an antagonistic protein mutant for cultivating a sweet potato composite virus disease (SPVD) resistant plant and application of the antagonistic protein mutant. The key pathogenic factor of the SPVD is the RNase3 protein coded by the sweet potato chlorotic stunt virus (SPCSV), and the RNase3 protein is used as an RNA silence suppressor (RSS) to destroy host immunity. The invention provides an SPCSV-RNase3 antagonistic protein mutant, which is derived from a natural antagonistic protein, namely a sweet potato trypsin inhibitor (IbSPLTI-a). According to the present invention, a compound structure model (such as construction through AlphaFold2) of wild type IbSPLTI-a and RNase3 is analyzed, and the IbSPLTI-a is modified by using a protein directed evolution strategy, particularly by using a hotspot amino acid scanning and protein surface design strategy, such that the high-activity inhibition mutant (such as IbSPLTI-a-m0805) is successfully created and screened; in-vivo and in-vitro function verification shows that the binding affinity of the mutant (such as IbSPLTI-a-m0805) and SPCSV-RNase 3 is remarkably enhanced, and the mutant shows a virus accumulation inhibition effect superior to that of a wild type IbSPLTI-a. The invention also provides a nucleic acid sequence for coding the mutant, a plant expression vector containing the sequence, and a method for culturing an anti-SPVD transgenic plant (especially sweet potato) by using the mutant. The invention provides a core gene resource and a technical path for genetic improvement of SPVD and development of a novel antiviral protein preparation.

This invention relates to the field of bioengineering technology, specifically to an endogenous signal peptide element fused with the MF-αpro region and its applications. The endogenous signal peptide element is composed of an endogenous signal peptide from Pichia pastoris with the C-terminus fused with MF-αpro or with amino acids 38 to 51 of the MF-αpro region detached; the amino acid sequence of the MF-αpro region is shown in SEQ ID NO:8; the amino acid sequence of the MF-αpro region with detached amino acids 38 to 51 is shown in SEQ ID NO:10. This invention provides a complete solution from rational screening to functional verification to element optimization. The resulting highly efficient, patent-free signal peptide element is suitable for the production of industrial enzymes, pharmaceutical proteins, and diagnostic reagents.

The invention discloses a wheat grain quality detection and functional gene analysis method based on a hyperspectral imaging technology, and belongs to the technical field of agricultural detection. The method comprises the following steps: extracting a hyperspectral index; determining a wheat grain quality index as an artificial measurement value; a weight coefficient is given to each quality index, and the wheat quality phenotype is digitalized; extracting a characteristic wavelength, and establishing a prediction model of each index and a wheat quality value; gWAS, SNP site screening and co-localization analysis are performed on the hyper-spectral index and the chemical index as phenotypic characters and genotypes respectively, and candidate genes are obtained and subjected to functional verification. The method combines a hyperspectral technology with chemometrics modeling, and is suitable for real-time monitoring in wheat quality breeding and processing processes; the spectrum phenotype is combined with GWAS gene mining, the limitation that traditional quality detection only serves phenotype screening is broken through, a'nondestructive testing-gene positioning 'closed loop is formed, and a target gene is provided for wheat quality improvement.

The utility model discloses a multifunctional verification and targeted culture integrated device for breast milk source probiotics, and relates to the technical field of breast milk source probiotics. The device comprises a detection system and a liquid supply system, the detection system is composed of a plurality of culture bins, an experiment cavity and a detection groove, the liquid supply system comprises a plurality of liquid supply points, a peristaltic pump and a valve block which are sequentially connected, each liquid supply point comprises a square seat, an upper sealing shell and a liquid storage tank, and a weighing sensor is arranged at the top of each square seat. According to the utility model, the verification module and the targeted culture module of each function of a bacterial strain are integrated, and the liquid supply system is designed, so that liquid can be supplied uniformly, the functional verification and culture efficiency of probiotics is improved, the research and development period is shortened, and the liquid supply amount is detected by designing a novel liquid supply point and in a weighing manner; and no interference exists between the liquid supply pipe and the liquid storage tank, so that the liquid supply accuracy is greatly improved, and the subsequent verification and culture effects are improved.

The invention discloses truncated high-activity cytosine deaminase as well as a related biological material and application thereof, and belongs to the technical field of protein. In order to improve the base editing efficiency and range, the invention provides a 15th-150th protein with an amino acid sequence of SEQ ID NO.2 and application of the 15th-150th protein as cytidine deaminase. The invention also provides application of the protein and related biological materials thereof in single base editing. According to the application, metagenome big data is utilized, comprehensive and systematic information mining analysis is carried out on novel cytidine deaminase, and a protein L91 which is highly similar to a known DddA protein structure but low in sequence similarity is selected from the novel cytidine deaminase for functional verification. The protein L91 has high catalytic activity as cytidine deaminase, is not limited by target sequence preference, can be widely applied to gene editing in vitro and in cells / tissues, and has good application potential and development value.

This invention belongs to the field of rice molecular breeding technology and discloses genes regulating cold and salt tolerance in rice seedlings. SOP17 Its applications. This gene has the nucleotide sequence shown in SEQ ID NO.1, and the amino acid sequence of the protein it encodes is shown in SEQ ID NO.2. This invention obtained a seedling-stage cold-resistant mutant of indica rice 9311 through EMS mutagenesis. sop17-1 The clone was obtained by analyzing the Mutmap. SOP17 Genes. Constructed using CRISPR / Cas9 gene editing technology. SOP17 Gene knockout vector, to obtain SOP17 Homozygous mutant lines. Functional validation showed that... SOP17 SOP17 This gene simultaneously negatively regulates cold and salt tolerance in rice seedlings. Knocking out this gene significantly enhances the tolerance of rice seedlings to low temperature (8°C) and salt (150mM NaCl). This invention provides new gene resources and molecular targets for breeding stress-resistant rice varieties.

The application discloses a rice gene expression and function verification integrated detection device, which comprises a box body, a liquid nitrogen freezing assembly for freezing rice is installed in the box body, a gene extraction assembly for extracting rice genes is installed in the box body, and a gene detection assembly for detecting and verifying rice genes is installed at the top of the box body; a plurality of steps such as liquid nitrogen freezing, sample grinding, nucleic acid extraction, purification and qPCR detection are integrated, full-process automatic operation is realized through a control module, manual frequent intervention is not needed, human operation errors are greatly reduced, the accuracy and repeatability of detection results are improved, a high-concentration saline solution is combined, a low-salt elution process design is adopted, automatic magnetic bead adsorption and cleaning processes are combined, interference substances such as proteins and impurities in samples are effectively removed, the nucleic acid purity can reach more than 95%, the recovery rate can reach more than 85%, and the reliability of gene expression and function verification results is ensured.

The invention discloses a Grin2b stably transfected cell strain, a construction method and application thereof and a method for screening pollutant high-throughput neurotoxicity or learning and memory disorders, and belongs to the technical field of biologication.The construction method comprises the following steps that 1, cells are cultured; (2) constructing a plasmid vector, wherein the plasmid vector comprises a Grin2b coding sequence; (3) virus coating and virus collection; (4) testing the titer of the lentivirus; (5) stable plant screening; and (6) carrying out stable plant transformation function verification. The Grin2b stably transfected cell strain based on the Grin2b related ion channel constructed by the invention provides a brand new technical platform for high-throughput neurotoxicity recognition, learning memory screening and mechanism research of organic pollutants and degradation products thereof.

This invention belongs to the field of genetic engineering technology, specifically involving TaSCAMP17 The application of its recombinant overexpression vector in improving plant salt tolerance, the TaSCAMP17 The nucleotide sequence is shown in SEQ ID NO.1, and the amino acid sequence is shown in SEQ ID NO.2. This invention will... TaSCAMP17 Functional validation was performed in Arabidopsis thaliana after overexpression, and the results showed that overexpression... TaSCAMP17 The salt tolerance of wild-type Arabidopsis thaliana is significantly improved, therefore TaSCAMP17 It plays a positive regulatory role in wheat salt tolerance, providing a theoretical basis and gene resources for wheat molecular breeding, and is of great significance for obtaining salt-tolerant and high-yielding lines and improving wheat salt tolerance in the future.

The invention discloses a bergamene synthetase SaBERS as well as a coding gene and application thereof. The amino acid sequence of the bergamotene synthetase is as shown in SEQ ID NO. 2. According to the invention, the function verification is carried out on the bergamene synthetase SaBERS from sandalwood. It is found that the sandalwood-sourced enzyme can be successfully expressed in a prokaryotic expression system to generate recombinant protein, and in an in-vitro enzymatic reaction system, the enzyme catalyzes FPP to generate alpha-bergamoene and beta-bergamoene. The contents of the two compounds calculated according to the peak area respectively reach 62.3 ng / ul and 13.4 ng / ul. According to the invention, the enzyme can be massively prepared by utilizing escherichia coli, is applied to synthesis of in-vitro bergamocene, can be further applied to synthesis of alpha-bergamocene or beta-bergamocene, and has wide industrial practical prospects and large-scale development potential.

The present invention belongs to the field of biotechnology. Provided are a new effector nuclease and the use thereof. On the basis of discovered and studied Cas12a effector nuclease sequences, two new clustered regularly interspersed short palindromic repeats (CRISPR)-associated effector nucleases are obtained by means of design, screening, and functional verification. Specifically provided is a Cas protein. The Cas protein is Cas-kf8 or Cas-kf9, wherein the Cas-kf8 has an amino acid sequence as shown in SEQ ID No. 1, and the Cas-kf9 has an amino acid sequence as shown in SEQ ID No. 4. The provided Cas protein demonstrates nuclease activity in experiments and has broad application prospects.

The invention provides a method for verifying functions of Cre in oil-producing microalgae and application of the method in enhancing biosynthesis of eicosapentaenoic acid, and belongs to the technical field of genetic modification. According to the invention, functional verification of Cre in oil-producing microalgae is completed by a method of inserting an intron from an endogenous Phatr3J51169 gene of phaeodactylum tricornutum into a Cre gene, that is, a selection marker located between two loxP sequences can be removed, and a free plasmid transformation method more suitable for Cre removal is further developed. Based on a Cre / loxP system, the expression of EPA synthetic pathway genes in the oil-producing microalgae is enhanced to obtain an engineering algal strain with high yield of EPA, and compared with a wild type, the relative content of EPA can be increased by 24.7%.