101 results about "Gene conversion" patented technology

The invention provides a Chinese cabbage clubroot susceptibility related gene BrSWEET14a, a construction method and application, belongs to the technical field of plant genetic engineering, and particularly relates to a Chinese cabbage clubroot susceptibility related gene BrSWEET14a. The CDS sequence of the Chinese cabbage BrSWEET14a is as shown in SEQ ID No. 1 (sequence identifier number 1). The gene is transformed into Col-type arabidopsis thaliana through an agrobacterium flower immersion transformation method, a BrSWEET14a heterologous expression arabidopsis thaliana strain is obtained, and the result shows that T-DNA insertion mutant of the BrSWEET14a homologous gene Atsweet14 is more resistant to clubroot than that of the Col-type arabidopsis thaliana, and heterologous expression of Chinese cabbage BrSWEET14a leads to reduction of disease resistance of arabidopsis thaliana to plasmodiophora brassicae. Therefore, the Chinese cabbage BrSWEET14a is closely related to the clubroot disease, and 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 and have a good application prospect.

A protein capable of inhibiting type I fimbriae of Escherichia coli (E. coli) is identified, where the protein is named as Type I fimbrial repressor (TIFR) protein, and is encoded by a TIfr gene with an ID of GW576_23830. A method is further provided for inhibiting the growth of type I fimbriae of an E. coli strain, in which the TIfr gene is transformed into the strain.

The invention provides an efficient and stable nori pythium gene transformation method which comprises antibiotic concentration screening, protoplast preparation, transformation system optimization, positive transformant screening and transformant phenotype and gene expression stability analysis, and the evaluation method is the nori pythium gene transformation method disclosed for the first time in China. The method provides a valuable tool for researching gene functions and pathogenic mechanisms of pythium aphanidermatum in the future.

The invention discloses an application of a TaAP2-4 gene in regulating and controlling starch content of wheat grains, which is used for enhancing the expression quantity of the TaAP2-4 gene to improve the starch content of the wheat grains and weakening the expression quantity of the TaAP2-4 gene to reduce the starch content of the wheat grains. The TaAP2-4 gene is located on a chromosome 4 of a wheat genome, and three copies, namely TaAP2-4-A, TaAP2-4-B and TaAP2-4-D, are formed on the chromosome 4. According to the invention, a Crispr / Cas9 gene editing vector for knocking out the TaAP2-4 gene is constructed by adopting a reverse genetics method, the TaAP2-4 gene is successfully knocked out in a wheat material by adopting an agrobacterium-mediated method, and the starch content of a knocked-out mutant is reduced, so that the condition that the TaAP2-4 gene plays a positive regulation role in a wheat grain starch synthesis process is known. In practical application, directional knockout mutation of the genome level TaAP2-4 gene can be achieved through a gene transformation process, the gene knockout mutation method can be used for verifying functions of the TaAP2-4 gene, and materials are provided for mining other starch synthesis related genes.

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

The invention relates to the field of transformation methods and application thereof, in particular to an agrobacterium tumefaciens-mediated pea leaf injection transient transformation method and application thereof, which comprises the following steps: introducing an exogenous gene into pea leaf cells through an injection method by using an agrobacterium tumefaciens-mediated gene transformation system in pea leaves, and realizing transient expression of the exogenous gene; according to the pea leaf injection transient transformation system based on agrobacterium tumefaciens mediation provided by the invention, through an optimized technical scheme, the problems of low transformation efficiency, unstable exogenous gene expression, high operation complexity and the like in the prior art are solved, and remarkable beneficial effects are brought to plant gene function research and commercial application; through an efficient, stable, simple and convenient pea leaf instantaneous transformation technology, many problems in the prior art are solved, the efficiency of gene function research and commercial application is improved, and a powerful technical support is provided for further development of plant molecular biology and genetic engineering technologies.

The invention discloses a methionine nutrition selection marker saccharomyces cerevisiae vector as well as a construction method and application thereof, and relates to the field of genetic engineering and synthetic biology, a methionine MET17 promoter and a coding region sequence are obtained through gene artificial synthesis, a leucine promoter and a coding region sequence in a pAG415-GAL1-ccdB vector are replaced by homologous recombination, and thus the methionine nutrition selection marker saccharomyces cerevisiae vector is obtained. A pAG415-GAL1-MET-ccdB expression vector is constructed and obtained, a ccdB gene is replaced by a green fluorescent protein reporter gene GFP sequence to be integrated into the pAG415-GAL1-MET-ccdB vector, a pAG415-GAL1-GFP-MET vector is constructed and obtained, the obtained expression vector can be used for screening exogenous gene transformants in a methionine deficient culture medium, meanwhile, green fluorescent GFP tag protein carried in the vector has the advantages that the expression vector can be applied to screening of the methionine deficient culture medium, and the expression efficiency of the methionine deficient culture medium is improved. And more application space is provided for carrying out subcellular localization, immunoprecipitation and other downstream inheritance and molecular biology analysis on a target gene in wine brewing cells.

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.

This invention discloses the application of the SPL23 gene in regulating the number of tassel branches in maize, including: applying a miR529-insensitive form of the SPL23 gene to regulate the number of tassel branches in maize. The method for increasing the number of tassel branches in maize is characterized by: constructing a vector containing a miR529-insensitive form of the SPL23 gene; and transforming the constructed miR529-insensitive form of the SPL23 gene into maize to obtain maize plants with an increased number of tassel branches. This invention, by constructing miR529-insensitive SPL23 plants, enables the SPL23 gene to resist miR529 degradation and thus undergo overexpression, significantly promoting tassel development and increasing the number of tassel branches, providing an effective approach for the genetic improvement of maize varieties with fewer male tassels.

The invention discloses a construction method and application of a sphingomonas ncd2 gene overexpression strain, and the ncd2 gene overexpression strain is formed by introducing an ncd2 gene into sphingomonas through a recombinant plasmid or integrating the ncd2 gene onto a chromosome of sphingomonas through a genetic engineering means. The construction method comprises the steps of designing an expression vector of the ncd2 gene, cloning the ncd2 gene, transforming sphingomonas, carrying out resistance screening, verifying expression of the ncd2 gene and optimizing culture conditions, after the ncd2 gene is applied to the broiler chicken, the intestinal microbial flora composition and the intestinal histological structure can be improved, meanwhile, the contents of total nitrogen, uric acid and ammonium nitrogen in the chicken manure of the broiler chicken can be remarkably reduced, and the yield of the broiler chicken is improved. The content of nitrate nitrogen is obviously increased, so that the slaughtering rate can be increased, and the economic benefit is further increased.

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.

The invention relates to the field of molecular biology and plant genetic engineering technology research, in particular to application of a sweet potato small peptide family gene IbCLE25 in regulation and control of drought resistance and salt tolerance. The nucleotide sequence of the IbCLE25 gene is as shown in SEQ ID NO. 1. The sweet potato IbCLE25 gene is transformed into a sweet potato variety Jiuzhou 25 to obtain an overexpressed sweet potato plant, and the result shows that compared with a wild type, after the sweet potato with the overexpressed IbCLE25 gene is stressed by drought and salt, the root length and fresh weight of the sweet potato are obviously higher than those of a control group, which indicates that the sweet potato IbCLE25 participates in the regulation and control of the drought resistance and salt resistance of the sweet potato, and the sweet potato IbCLE25 gene can be applied to the control of the drought resistance and salt resistance of the sweet potato. Reference is provided for stress resistance improvement and new variety breeding of sweet potatoes.

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.

The invention discloses a Trichoderma reesei engineering strain for producing trans-aconitic acid as well as a construction method and application of the Trichoderma reesei engineering strain, and relates to the technical field of biological engineering. The construction method comprises the following steps: transforming a coding gene of mitochondrial tricarboxylic acid transport protein and a coding gene of aconitic acid isomerase into a trichoderma reesei host bacterium, and carrying out gene recombination construction to obtain the trichoderma reesei engineering strain, the amino acid sequence of the mitochondrial tricarboxylic acid transporter is as shown in SEQ ID NO. 1; the amino acid sequence of the aconitic acid isomerase is as shown in SEQ ID NO. 2 or SEQ ID NO. 3. A new production strain is provided for microbial fermentation production of trans-aconitic acid, a large amount of trans-aconitic acid can be produced through direct fermentation by using common carbon sources such as glucose or cellulose, and the highest yield can reach 26.5 g / L.

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 present invention provides a sweet potato salt-tolerance-related gene IbCHR10 and its application, belonging to the field of molecular biotechnology. The present invention provides a sweet potato salt-tolerance-related gene IbCHR10, which lacks a conserved DST element region downstream of the 3' UTR region, resulting in stable mRNA after translation, thus filling a gap in the prior art. The present invention successfully cloned IbCHR10 from sweet potato and confirmed that the gene is expressed at higher levels in sweet potato fibrous roots than in other tissues. The gene was transformed into sweet potato using Agrobacterium-mediated transfection, and it was found for the first time that overexpression of IbCHR10 enhances salt tolerance in sweet potato, further demonstrating that the IbCHR10 gene is related to sweet potato salt tolerance. This indicates that the present invention's gene IbCHR10 has broad application prospects in sweet potato stress resistance research.

The application discloses a Phyllostachys edulis PeBBX19 gene, a coding protein and application thereof, and belongs to the technical field of plant genetic engineering.The nucleotide sequence of the Phyllostachys edulis PeBBX19 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 an overexpression vector of the Phyllostachys edulis PeBBX19 gene, transforms the constructed overexpression vector of the Phyllostachys edulis PeBBX19 gene into Arabidopsis, and cultivates, screens and obtains a transgenic Arabidopsis strain with delayed flowering time.The application discloses the function of the PeBBX19 gene for the first time, the transgenic Arabidopsis plant has delayed flowering time after the PeBBX19 gene is transformed into the Arabidopsis plant, obviously shows a late flowering phenotype, and the number of rosette leaves is increased; and the expression amount of a downstream regulation gene FT is significantly reduced.

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.

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 present invention provides, among other things, a transgenic chicken. In some embodiments, the transgenic chicken comprises a B cell having an endogenous immunoglobulin heavy chain locus comprising: (a) a functional immunoglobulin heavy chain gene comprising a nucleic acid encoding a heavy chain variable domain having a length of CDR3 in the range of 30 to 60 amino acids and containing at least two cysteine ​​residues; and (b) a plurality of pseudogenes operably linked to the functional immunoglobulin heavy chain gene and, by gene conversion, providing a nucleotide sequence to the nucleic acid encoding the heavy chain variable domain of (a), wherein the pseudogenes are located upstream or downstream of the functional immunoglobulin heavy chain gene.

Disclosed herein are systems, devices, and methods for identifying recombinant variants (such as gene conversion variants) of genes such as RHD gene and RHCE gene, the copy numbers of recombinant variants, and gene variant status (for example, heterozygous or homozygous). In some embodiments, the disclosed systems, devices, and methods include steps of receiving sequence reads which align to a RHD gene or a RHCE gene, estimating a combined copy number of a RHD gene and a RHCE gene, estimating copy numbers of a RHD-specific base and a RHCE-specific base at each of a plurality of pre-determined differentiating sites of the RHD gene and the RHCE gene, and calculating a probability of a RHCE*CE-D(2)-CE gene conversion in the nucleic acid sample.

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 application provides a Chinese cabbage root knot disease related gene BrSWEET14a, a construction method and application, belongs to the technical field of plant genetic engineering, and particularly relates to the following technical scheme. The CDS sequence of the Chinese cabbage BrSWEET14a is shown as SEQ ID No. 1. The gene is transformed into Col-type Arabidopsis thaliana through an Agrobacterium flower dipping transformation method, and a BrSWEET14a heterologous expression Arabidopsis thaliana strain is obtained. It is found that the T-DNA insertion mutant of the BrSWEET14a homologous gene Atsweet14 is more resistant to root knot disease than the Col-type, and the heterologous expression of the Chinese cabbage BrSWEET14a leads to the decrease of the disease resistance of Arabidopsis thaliana to the root knot fungus. It can be seen that the Chinese cabbage BrSWEET14a is closely related to the root knot disease, and the gene and the homologous genes of the gene in other cruciferous plants can be applied to the breeding of cruciferous plants and other horticultural plants, and have a good application prospect.

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.