42 results about "Gene conversion" patented technology

This invention discloses the application of the wheat gene TaPSS2 in improving plant drought resistance, belonging to the field of functional genomics. The nucleotide sequence of the wheat gene TaPSS2 is shown in SEQ ID NO: 2. Transforming this gene into the model plant Arabidopsis thaliana can significantly improve the drought resistance of Arabidopsis. If the wheat gene TaPSS2 is transformed into wheat, rice, maize, cauliflower, and other plants, it is possible to obtain new germplasm with drought-resistant characteristics, which is of great significance for the breeding of superior crop varieties and their widespread application in production.

The invention belongs to the technical field of biology, and relates to application of a ZmCASPL11 gene in improving heat resistance of plants. The ZmCASPL11 gene encodes a protein of a membrane protein (CASPL) family related to a Kjeldahl band structure, the gene expression level of the ZmCASPL11 gene under high-temperature stress is remarkably increased, and a mutant zmcaspl11 of the gene shows a high-temperature sensitive phenotype. The ZmCASPL11 gene disclosed by the invention 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 ZmCASPL11 gene to heat stress is remarkably enhanced. According to the invention, more theoretical bases are provided for corn heat resistance research, and gene resources and biotechnological means are provided for subsequent cultivation and germplasm resource innovation of new heat-resistant corn strains.

Provided herein is a transgenic chicken comprising a genome having an endogenous immunoglobulin heavy chain gene in which the IgY CH1 coding sequence is functionally deleted. In certain embodiments, in B cells of the chicken the endogenous immunoglobulin heavy chain gene comprises: (a) a functional immunoglobulin heavy chain gene comprising, in operable linkage, a nucleic acid encoding an antibody heavy chain variable domain and a constant region coding sequence in which the IgY CH1 coding sequence is functionally deleted; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the variable domain of (a), wherein the pseudogenes are upstream of the nucleic acid encoding an antibody heavy chain variable domain of (a).

This disclosure provides, among other things, a transgenic animal that uses gene conversion for antibody diversification, comprising B cells in which the endogenous immunoglobulin heavy chain locus comprises: (a) a functional immunoglobulin heavy chain gene comprising a nucleic acid encoding an autonomous heavy chain (AHC) variable domain; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the AHC variable domain of (a), wherein the pseudogenes are upstream or downstream of the functional immunoglobulin heavy chain gene.

This disclosure provides, among other things, a transgenic chicken. In some embodiments, the transgenic chicken comprises B cells in which the endogenous immunoglobulin heavy chain locus comprises: (a) a functional immunoglobulin heavy chain gene comprising a nucleic acid encoding a heavy chain variable domain in which the CDR3 is in the range of 30-60 amino acids in length and comprises at least 2 cysteine residues; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the heavy chain variable domain of (a), wherein the pseudogenes are upstream or downstream of the functional immunoglobulin heavy chain gene.

This invention belongs to the field of biotechnology and relates to a... ZmCASPL11 The application of genes in improving plant heat tolerance. ZmCASPL11 The gene encodes a protein belonging to the Casparian band structure family of membrane proteins (CASPL), whose expression level is significantly upregulated under high temperature stress. Mutants of this gene... ZmCASPL11 It exhibits a high-temperature sensitive phenotype. The present invention describes... ZmCASPL11 Genes were transformed into Arabidopsis thaliana and their function was validated. Compared with wild-type Arabidopsis thaliana, overexpression of [the gene] showed [significant improvement]. ZmCASPL11 The gene significantly enhances the tolerance of Arabidopsis thaliana plants to heat stress. This invention provides more theoretical basis for research on heat resistance in maize, and offers genetic resources and biotechnological means for the subsequent breeding of new heat-resistant maize lines and the innovation of germplasm resources.

This disclosure provides, among other things, strategies for minimizing antibody diversification in a transgenic animal that uses gene conversion for antibody diversification. In some embodiments, the animal may comprise a genome comprising an endogenous immunoglobulin light chain locus comprising: (a) a functional immunoglobulin light chain gene comprising a nucleic acid encoding a light chain variable region; and (b) a plurality of pseudogenes that are operably linked to the functional immunoglobulin light chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding a light chain variable region, wherein the pseudogenes are upstream or downstream of the functional immunoglobulin light chain gene and encode the same amino acid sequence as the light chain variable region of the functional immunoglobulin light chain gene of (a). In other embodiments, the locus may have a tandem array of coding sequences for the light chain.

The invention discloses a capsicum annuum cell cycle gene CaTAM and application thereof. The DNA sequence of the capsicum annuum cell cycle gene CaTAM is as shown in SEQ ID No.1. The gene is transformed into Columbia type arabidopsis thaliana through an agrobacterium flower immersion transformation method, a CaTAM heterologous expression arabidopsis thaliana strain is obtained, and it can be known that the heterologous expression of CaTAM can cause early bolting time of arabidopsis thaliana, increase of the number of leaves, decrease of the length and width of the leaves and increase of the root length. The expression of CaTAM of capsicum is silenced through a VIGS technology, a plant with reduced CaTAM expression is obtained, and it can be known that due to silencing of CaTAM, part of pollen cells exit meiosis in advance, secondary meiosis is not carried out, and large pollen grains are formed. It shows that CaTAM plays an important regulation and control role in meiosis, bolting time regulation and control, leaf growth, primary root development and the like, and the gene can be applied to breeding of solanaceae vegetables and other horticultural plants and has a good application prospect.

The invention discloses a ceramic cultural and creative product design and generation method. The method comprises the following steps: culture gene extraction: extracting dominant culture genes and recessive culture genes from target cultural heritage; user demand analysis: carrying out demand investigation on a target user based on a rooting theory, constructing a user demand emotion profile through an emotion calculation technology, and obtaining quantization intensity of user emotion dimensions; the material character elements are converted into external characterization of a product through a symbol conversion method, space narrative genes are converted into interactive logic or experience processes of the product, quantitative mental concepts and user demand emotion profiles are subjected to matching analysis, and emotion resonance design is conducted through an emotion migration method; and product generation and resource management: generating a ceramic cultural and creative product design scheme based on the translated design elements, and monitoring and evaluating the resource pushing condition in the design process.

The invention discloses a promoter PSCBV-FN39 and application of the promoter PSCBV-FN39. The nucleotide sequence of the promoter PSCBV-FN39 is as shown in SEQ ID NO: 1. The promoter disclosed by the invention is a rhizome tissue high-expression promoter, can be applied to gene transformation of various crops, preparation of transgenic plants and breeding of the transgenic plants, and has an important value for promoting acquisition of plant strains with high quality, high yield, high disease resistance, high stress resistance and other special characters; the development of a modern genetic breeding technology can be effectively promoted.

The invention provides application of a TaPAL8 gene in positive regulation of phenolic acid content of wheat, and belongs to the technical field of gene engineering. Wide targeted metabolome detection is carried out on wheat whole growth period materials by utilizing an HPLC-MS / MS technology, specific accumulation of various phenolic acids in stalks is highly related to expression of multiple phenylpropane pathway genes, one gene is annotated as phenylalanine ammonialyase, the gene number is TraesCS6D01G212200, and the gene is named as TaPAL8. According to the embodiment of the invention, the gene TaPAL8 is transformed into a wheat variety Fielder to obtain a transformed plant with overexpression TaPAL8, and in the overexpression mutant, the content of various acyl modified quinic acid substances in stalks and grouted grains is remarkably increased.

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 application relates to the field of plant genetic engineering, and specifically discloses a high-efficiency gene transformation system based on the stem tip meristem of a yellowing seedling of Zhongmian 113. The core components of the system include L1 medium suitable for Zhongmian 113, Agrobacterium GV3101 carrying a target gene, a special resuspension solution containing acetosyringone and Silwet-L77, and special primers for overexpression and gene editing; and the preparation method is as follows: seed disinfection and germination are performed to obtain a yellowing seedling, the stem tip is cut in an X shape to expose the meristem, Agrobacterium infection is performed, and then dark culture and light culture are performed, and after transplanting and cultivation to the true leaf stage, positive plants are screened and identified through PCR; the system can be used for gene transformation of Zhongmian 113 and improvement of high yield, high quality, disease resistance and the like, and has the advantages of strong adaptability, high transformation efficiency and short cycle; in addition, the preparation method of the application does not need callus regeneration, is simple and convenient to operate, has a standardized process, is good in repeatability and stability, and can quickly realize transgenic seed acquisition.

This invention discloses the application of the jujube ZjUVI4 gene in rice grain shape improvement, belonging to the field of plant genetic engineering technology. The invention cloned the coding region sequence of ZjUVI4 from the jujube variety 'Dongzao' using PCR. The full-length sequence is 774 bp, encoding 257 amino acids. The nucleotide sequence of the coding region of the ZjUVI4 gene is shown in SEQ ID NO:1. Then, an overexpression vector was constructed, and transgenic experiments were conducted using the rice variety 'Zhonghua 11' as the transformation recipient. The ZjUVI4 gene was heterologously overexpressed in rice, and its biological function in the positive regulation of rice grain shape was investigated. This invention reveals the biological function of the jujube ZjUVI4 gene in positively regulating seed size. By transforming the ZjUVI4 gene into rice, the transgenic rice lines showed a significant increase in grain length and thousand-grain weight, while grain width remained largely unchanged, providing a new gene resource for rice grain shape improvement and high-yield, high-quality breeding.

The application discloses a Phyllostachys edulis PeSVP gene, a coding protein and application thereof, and belongs to the technical field of plant genetic engineering.The nucleotide sequence of the PeSVP gene disclosed by the application is shown as SEQ ID NO.1, and the amino acid sequence of the coding protein is shown as SEQ ID NO.2.The application constructs a PeSVP gene overexpression vector of Phyllostachys edulis, transforms the constructed PeSVP gene overexpression vector of Phyllostachys edulis into Arabidopsis thaliana, and cultivates, screens and obtains a transgenic Arabidopsis thaliana strain with delayed flowering time.The application discloses the function of the PeSVP gene for the first time, the transgenic Arabidopsis thaliana plant has delayed flowering time after the PeSVP gene is transformed into the Arabidopsis thaliana plant, shows an obvious late-flowering phenotype, and the number of rosette leaves is increased;the expression amount of a downstream regulation gene FT is significantly reduced.The application can be used in molecular breeding of plant flowering period regulation, and has important significance for cultivating new plant germplasm with higher biological yield.

This invention discloses Shprh The application of gene-targeted therapies in the preparation of drugs that enhance immunity falls within the field of molecular biology. This invention utilizes the Cre-loxp system to construct a conditional knockout mechanism in B cells. Shprh Genetic Shprh fl / fl Cd19 Cre / + Mice. Specific knockout in B cells was found. Shprh Following gene therapy, the frequency of gene conversion in mice was significantly increased, and the diversity of antibodies in mice was enhanced, which is expected to further improve their immunity.

The application discloses a beta-lactamase and a gene for coding the beta-lactamase, and further discloses an expression cassette, a recombinant carrier, a recombinant cell or a recombinant bacteria, and a construction method of the recombinant bacteria, and further discloses the application of the beta-lactamase, the gene for coding the beta-lactamase, the expression cassette, the recombinant carrier, the recombinant cell or the recombinant bacteria, and the construction method of the recombinant bacteria in removing beta-lactam antibiotic pollution in the environment. The beta-lactamase gene is transformed into escherichia coli, so that the escherichia coli can hydrolyze beta-lactam antibiotics in the environment, including ampicillin, cefazolin, cefotaxime, imipenem and amikacin, and has great application value in eliminating beta-lactam antibiotic pollution in the environment.

The invention discloses a Chinese cabbage clubroot-resistant gene BrRLP30 and application thereof. The nucleotide sequence of the Chinese cabbage clubroot-resistant gene BrRLP30 is as shown in SEQ ID No.1. The invention further discloses a preparation method of the Chinese cabbage clubroot-resistant gene BrRLP30. After the endogenous BrRLP30 gene of the Chinese cabbage is silenced by a VIGS technology, the susceptibility of the plant to clubroot is increased, the disease index is obviously higher than that of a control group, and the root swelling symptom is aggravated. The gene is transformed into a Chinese flowering cabbage plant through an agrobacterium flower immersion transformation method to obtain a BrRLP30 heterologous overexpression strain, and a disease resistance identification result shows that the overexpression of BrRLP30 significantly enhances the resistance of the plant to clubroot. Therefore, the gene BrRLP30 is closely related to the clubroot disease, the gene and the homologous gene of the gene in other cruciferous plants can be applied to breeding of the cruciferous plants and other horticultural plants, the disease resistance of the plants can be improved, and the gene BrRLP30 has a good application prospect.

The invention relates to the technical field of competent escherichia coli cell preparation, and discloses a preparation method of competent escherichia coli cells for gene transformation, which comprises the following steps: step 1, strain activation; step 2, strain amplification culture; step 3, low-temperature suspension and washing: transferring the bacterial liquid into a pre-cooling centrifugal tube, standing on ice, then centrifuging, discarding supernate, adding a pre-cooled CaCl2 solution, gently resuspending, placing on ice, and then centrifuging; step 4, preparation of a competent suspension: removing the supernatant, adding a precooled CaCl2 solution containing glycerol, slightly suspending thalli, and standing on ice for 3-5 minutes to prepare a competent cell suspension; the mass fraction of glycerol in the CaCl2 solution containing glycerol is 15%; the concentration of CaCl2 is 0.05 mol / L; and step 5, sub-packaging and cryopreservation: sub-packaging the obtained competent suspension, and cryopreserving at-80 DEG C. According to the scheme, by optimizing the concentration of CaCl2 and glycerol in the suspension, the suspension temperature and the like, the conversion efficiency, the storage period and the cell membrane stability are effectively improved.

The application discloses a Trichoderma reesei engineering bacterium for secreting and expressing egg white lysozyme and a construction method thereof, and belongs to the field of molecular biology and biotechnology. The application firstly provides an egg white lysozyme mutant, and the amino acid sequence is shown in any one of SEQ ID NO. 2-4. The construction method of the Trichoderma reesei engineering bacterium for secreting and expressing egg white lysozyme comprises the following steps: taking Trichoderma reesei as a starting strain, and transforming an expression module into the Trichoderma reesei to construct the Trichoderma reesei engineering bacterium; the expression module comprises a promoter, a secretion peptide gene, an egg white lysozyme gene and a terminator which are sequentially connected. By transforming the coding gene of the egg white lysozyme into the Trichoderma reesei genome, the Trichoderma reesei engineering strain for secreting and expressing egg white lysozyme is successfully constructed. It is found that different egg white lysozyme variants can bring different effects, and when the Trichoderma reesei RUT-C30 is used as the starting strain, the effect is better than that of QM6a.

The invention discloses a method for producing an active rhBMP9 bone repair material based on a double-transgene bombyx mori silk gland biosynthesis system as well as a product and application of the active rhBMP9 bone repair material, and realizes cooperative expression of complex precursor protein Pro-mhBMP9 and precursor protein invertase hFurin in bombyx mori posterior silk glands. An hBMP9 / hFurin double-gene biosynthesis system capable of completing precursor protein shearing, maturation and functionalized silk material synthesis in vivo is established, the rhBMP9 / CaP / Sericin composite scaffold is further constructed by extracting silk protein of double-gene transformation bombyx mori, and the rhBMP9 / CaP / Sericin composite scaffold shows excellent bone repair and angiogenesis capacities in both the cell level and the animal level, and can be used for preparing a bone repair material. And a solid experimental and theoretical basis is provided for a functional protein-material-tissue regeneration integrated strategy based on a silk gland biosynthesis system.

The present application relates to the field of biotechnology, in particular to a white flower carum gene MaUGT79 and its application, the present application is based on white flower carum genome data and BSA resequencing data, cloning MaUGT79 gene, using real-time quantitative PCR technology to analyze the expression pattern under drought stress, and construct the recombinant prokaryotic expression vector to verify MaUGT79 gene function, it is found that transgenic escherichia coli can catalyze scopolin to produce scopolamine; construct plant overexpression vector and RNAi vector to transform the gene into white flower carum hairy root, verify MaUGT79 gene to participate in scopolin synthesis, has the function of resisting drought stress, the present application lays the foundation for revealing the drought tolerance mechanism of MaUGT79 gene, provides gene resources and theoretical basis for plant drought tolerance molecular breeding.

The invention discloses a plant insect-resistant related protein as well as a coding gene and application thereof. According to the invention, the full length of the SlJAIB14 gene is amplified from tomato, and two restriction enzyme cutting sites of BamHI and Sac I are introduced to the 5'and 3 'ends of the gene through primer sequences. The newly synthesized gene is connected with a pYPX158 plant expression vector containing double 35S promoters through a T4 DNA (Deoxyribose Nucleic Acid) ligase, so that a recombinant plasmid pYPX-S1JAIB14 containing a target gene S1JAIB14 is obtained. According to the invention, plasmids are introduced into agrobacterium tumefaciens (GV3101) by using an electric shock method. The SlJAIB14 gene is transformed into tomato by using an agrobacterium-mediated method, and the resistance of the transgenic plant to cotton bollworm is verified through a feeding experiment.

Microbes are transformed with psilocybin genes under the control of weak or medium level promoter to make low levels of psilocybin therein. Low dose, microdose and sub-microdose foodstuff are then made with such microbes.

Provided is a mutant polyhydroxyalkanoic acid synthase comprising an amino acid sequence which has at least 90% sequence identity with the amino acid sequence represented by SEQ ID NO: 1 and in which at least one among 11th, 23rd, 74th, 176th, 224th, 241st, 327th, 360th, 400th, 419th, 452nd, 484th, 485th, 509th, and 511th amino acids from the N-terminal is substituted with a specific amino acid, wherein the mutant polyhydroxyalkanoic acid synthase has a synthetic activity of a copolymerized polyhydroxyalkanoic acid including a 3-hydroxyalkanoic acid monomer unit having 8 or more carbon atoms.

The invention belongs to the technical field of gene engineering, and particularly relates to application and a method of an SPL9 gene in regulation and control of insect resistance of forest trees and cultivation of insect-resistant forest tree varieties. According to the application of the SPL9 transcription factor gene in regulating and controlling the insect resistance of forest trees, the sequence of the SPL9 gene is shown as SEQ ID NO: 1. The insect resistance of the forest plant with the SPL9 gene knocked out is remarkably improved in the forest plant transformed by using the SPL9 gene with the designed specific targeting coding sequence shown as SEQ ID NO: 1, and the insect resistance of the forest plant with the SPL9 gene knocked out is remarkably improved. According to the invention, the SPL9 transcription factor gene is knocked out from forest trees through a gene editing technology, so that the SPL9 transcription factor gene is further used for cultivating new varieties with improved insect resistance, and has great application value in molecular breeding.

The invention discloses an escherichia coli mutant, a construction method and application thereof, and a sorting and enriching method of membrane-free organelles, and relates to the technical field of microorganisms. The escherichia coli mutant is obtained by inactivating an minD gene on the basis of escherichia coli BL21 (DE3). The escherichia coli mutant provided by the invention can generate small cells, and the small cells can be used as a carrier for sorting and enriching membrane-free organelles. Specifically, after a gene for coding a bacterial phase change protein is transformed into an escherichia coli mutant for culture, sorting and enrichment of the membraneless organelle can be realized by collecting small cells in a bacterial solution, and a natural structure of the membraneless organelle can be reserved under an in-situ condition.

The invention discloses a yeast heterologous expression recombinant human collagen III and a preparation method thereof. The yeast heterologous expression recombinant human collagen III comprises an amino acid sequence as shown in SEQ ID No. 1. The preparation method comprises the following steps: cloning a nucleotide sequence as shown in SEQ ID No.2 into a vector to construct a recombinant expression vector plasmid I; the method comprises the following steps: cloning a nucleotide sequence shown as SEQ ID No.3 into a vector, and constructing a recombinant expression vector plasmid II; transfecting the recombinant expression vector plasmids I and II into host cells, and screening to obtain double-gene transformed recombinant bacteria; fermenting and culturing the double-gene transformation recombinant bacteria; and purifying the yeast heterologous expression recombinant human III type collagen. On the basis of retaining the hydrophobic structure and activity of the original human-derived III-type collagen, the stability and hydrophilicity of the protein structure are improved; a P4H sequence is introduced, so that the defect that a pichia pastoris expression system cannot be hydroxylated is overcome.

A method for efficient genetic transformation and gene editing of a Brassica crop mediated by Agrobacterium rhizogenes is provided. Genetic transformation of the Brassica crop and establishment of a gene editing system are achieved by Agrobacterium rhizogenes-mediated delivery of three developmental regulators (DRs), ZmWUS2, AtIPT, and AtPLT5. The Agrobacterium rhizogenes-mediated delivery of three DRs, ZmWUS2, AtIPT, and AtPLT5, may induce explants to form callus and then directly form buds. The method solves the problem of difficult root regeneration for the Brassica crop, and enables species with a poor root regeneration ability to achieve efficient transformation.