73 results about "Gene technology" patented technology

The invention discloses a site for stably expressing protein in a CHO cell gene NW023276806.1 and application of the site, and belongs to the technical field of biological genes. The site belongs to a fixed position in a CHO cell genome, different protein genes are introduced based on a micro-homologous end connection mechanism through a CRISPR / Cas9 tool, and stable expression is carried out. By adopting a site-specific integration method, a target gene is integrated to a stable expression area in a site-specific manner, repeated high-expression monoclonal screening is effectively avoided, and an MMEJ mechanism is introduced to integrate a donor fragment, so that the research and development time for constructing a stable expression cell strain in biological pharmacy can be effectively shortened, and the cost is reduced.

The invention belongs to the technical field of genes, and discloses a method for efficiently expressing a foreign protein based on an NC048604-1 site in a CHO cell genome. A site for stably expressing protein in the CHO cell genome is located in the 94142000 to 94148000 basic group range of the CHO cell genome NC048604-1, and the nucleotide sequence of the site is as shown in SEQ ID NO: 1. According to the invention, different protein genes are introduced at fixed positions in a CHO cell genome, and stable expression is carried out.

The invention discloses an annotation method for analyzing a genome centromere region structure, and belongs to the technical field of biological genes. And prerequisites are needed. The method comprises the following steps: firstly, analyzing repetitive monomers of a centromere sequence based on a heuristic strategy, and judging the centromere sequence; when the serial repetition units are identified, an adjacent matrix is obtained based on serial repetition unit information, the analysis problem of the HORs is converted into a graph theory problem of searching a specific structure in the adjacent matrix, and a sliding window strategy is combined to refine the structure analysis result of the HORs. Compared with the prior art, the method has the advantages that the length of the tandem repeat unit can be detected more accurately, and the defect that two continuous tandem repeat units are identified as a single tandem repeat unit by TRASH is effectively overcome. The method has an obvious efficiency advantage. When a human genome centromere area is analyzed, the method is 10 times faster than a current method while the precision is guaranteed.

The invention belongs to the technical field of genes, and discloses a method for efficiently expressing foreign protein based on an NC048599.1 site in a CHO cell genome. A site for stably expressing a protein in a CHO cell genome is located in a range of 69195000 to 69199000 of a CHO cell gene NC048599.1 with a sequence as shown in SEQ ID NO: 1 (Sequence Identifier Number 1), and the sequence of the CHO cell gene NC048599.1 is shown in SEQ ID NO: 1. Different protein genes are introduced at fixed positions in a CHO cell genome, and stable expression is carried out.

The invention relates to the technical field of genes, in particular to preparation of a Trmt61a enzyme activity deleted zebrafish model, and the Trmt61a enzyme activity deleted zebrafish model is used for preparing a zebrafish strain which stably inherits trmt61aD181A-P2A-EGFP knock-in zebrafish strain. According to the invention, trmt61a D181A mutation site specificity is introduced into a zebra fish genome by using a CRISPR / Cas9 technology, and a zebra fish model with Trmt61a catalytic activity deficiency and tRNA m1A modification level significantly reduced is obtained. The zebrafish model fills the existing research blank, provides an available in-vivo research model for biological development regulation mechanism research related to tRNA m1A epigenetic modification and pathological mechanism analysis of diseases (such as malignant tumors and hematopoietic dysfunction), and can be directly used for exploring the core effect of the epigenetic modification in development and diseases.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a chrysanthemum indicum CzNAC1 gene and application thereof, and the nucleotide sequence of the gene is as shown in SEQ ID NO.1. According to the invention, transcriptome data is screened and cloned to the drought-responsive CzNAC1 gene, a carrier is constructed, and a transgenic plant is cultivated, so that the cultivation of a specific variety of transgenic chrysanthemum indicum, drought resistance conditions, economic and social benefits increase, stress-resistant genes are explored, gene resource mining and cross-species application are further promoted, and the application prospect is wide. And a good basis is provided for subsequent gene technology research.

The invention discloses a gene OsNST2 for transporting nucleotide sugar and application of the gene OsNST2. A nucleotide sequence of a coding region of the gene is as shown in SEQ ID No. 1, and an amino acid sequence coded by the gene is as shown in SEQ ID No. 2. It is found for the first time that the growth, development and yield of rice are affected by the OsNST2 by regulating and controlling formation of rice cell walls, an OsNST2 gene knock-down strain is created through an RNAi technology, an OsNST2 gene overexpression strain is obtained through an overexpression transgenic technology, it is verified that the OsNST2 gene is a new gene for regulating and controlling formation of the rice cell walls, and a new gene resource is provided for high yield of rice.

The invention discloses a transcription factor TgWRKY65 of torreya grandis as well as a coding gene and application of the transcription factor TgWRKY65. The amino acid sequence of the transcription factor TgWRKY65 is shown as SEQ ID NO.2, and the nucleotide sequence of the coding gene is shown as SEQ ID NO.1. The invention further discloses a preparation method of the transcription factor TgWRKY65. According to the invention, a homologous expression material is constructed through cloning of the TgWRKY65 gene, a transgenic technology and gene silencing, and the application prospect in the aspect of increasing the content of the torreya grandis catechin is great; when the coding gene is over-expressed, the content of catechin is increased; when the coding gene is silenced, the content of catechin is reduced.

This invention belongs to the field of gene technology for disease control. This invention provides a method for preparing biocontrol engineered bacteria using *Trichoderma harzianum* as a dsRNA vector, comprising the following steps: determining the target gene based on the pathogenic fungus; using the target gene as an arm sequence, and centering on the VdTublin intron sequence of the *Verticillium dahliae* endogenous gene, connecting two identical arm sequences to both ends of the intron in a forward and reverse direction, respectively, to construct the target sequence; and transferring the target sequence into *Trichoderma harzianum* using ATMT to obtain engineered *Trichoderma harzianum* bacteria. The engineered strain obtained by this invention has a good inhibitory effect on the growth and pathogenicity of the corresponding fungus.

The invention relates to a rice gene OsMYB63L and application of the rice gene OsMYB63L in regulation and control of spike branching and yield, and belongs to the technical field of plant gene breeding, the nucleotide sequence of the gene OsMYB63L is as shown in SEQ ID NO.1, the amino acid sequence of protein coded by the gene is as shown in SEQ ID NO.2, overexpression of the gene OsMYB63L promotes development of the spike branching of rice, then the yield of rice is increased, a transgenic technology is utilized, and the yield of the rice is increased. Overexpression is carried out on the rice gene OsMYB63L, and a rice plant with increased ear branch number and increased yield can be obtained. The invention discovers and verifies that the rice gene OsMYB63L has the function of regulating and controlling the ear branching number and the yield of rice. And a new gene resource and a technical path are provided for molecular breeding of high-yield rice.

The invention belongs to the technical field of biochemical engineering and genes, and discloses a preparation method and application of a high-yield mutant strain for improving the yield of carotene in schizochytrium limacinum, and the preparation method comprises the following steps: carrying out ARTP mutagenesis on a bacterial suspension in a logarithmic phase of schizochytrium limacinum HX-308 for 60 seconds, and washing the mutated thalli with 1M sorbitol to obtain an ARTP mutagenesis bacterial suspension; the method comprises the following steps: diluting a bacterial suspension by 100 times with a 1M sorbitol solution, coating the diluted bacterial suspension on a primary screening flat plate containing 25 [mu] M of fluridone, selecting a single colony with the diameter of more than or equal to 0.2 cm for fermentation culture, breeding a schizochytrium limacinum carotene accumulation dominant strain, and determining that the carotene yield of a schizochytrium limacinum mutant strain is more than 1.5 times higher than that of wild type schizochytrium limacinum HX-308. The mutant strain is a high-yield mutant strain for the carotene yield of the schizochytrium limacinum. According to the method, fluridone is used as a screening agent, the specificity is good, high-yield mutant strains can be rapidly and preliminarily screened, and the method is simple and efficient.

The present invention relates to a single nucleotide polymorphism detection primer for the sheep BMPR1B gene and its application, belonging to the field of gene technology. The SNP molecular marker corresponding to the primer of the present invention is located on chromosome 6 of the BMPR1B gene, and the primer is used for sheep marker-assisted selection breeding. The present invention provides a plurality of SNP molecular marker sites related to sheep reproductive traits, and determines the genotype of each site by directly sequencing the PCR product using sheep blood genomic DNA as a template. The present invention can be used as an effective molecular marker to improve sheep reproductive traits. The detection method is simple, rapid, and highly accurate, which can improve the breeding efficiency of sheep. It has high application value and provides more ways for the screening research of sheep reproductive traits.

The invention provides a gene for regulating and controlling the shape of a tomato leaf and application thereof, and a construction method of a recombinant expression vector is characterized in that an SlPUB26 gene is inserted into a vector containing a strong promoter by utilizing an overexpression transgenic technology. Transgenic plants with different gene expression level up-regulation multiples are obtained through agrobacterium tumefaciens-mediated transformation, so that single plants with different leaf phenotype change degrees are obtained. According to the method, a precise targeting sequence does not need to be designed, the miss-target hidden danger does not exist, and single plants with different SlPUB26 gene expression levels can be obtained only by detecting the gene expression quantity, so that the single plants with different leaf shape change degrees are obtained. The technical operation difficulty is low, and the method is more suitable for non-professional molecular breeding teams or grass-roots agricultural scientific research institutions. The higher the up-regulation multiple of the expression level of the SlPUB26 gene is, the lower the length-width ratio of the tomato leaf is, so that the shape of the tomato leaf can be quickly and accurately adjusted by adjusting the expression level of the SlPUB26 gene.

The invention discloses an FvePREs transcription factor and application thereof in promoting curling of strawberry petals, in a general agricultural cultivation technology, the petals are curled by gibberellin (GA3) or a growth regulator for actively inducing the curling of the petals, and excessive hormone may cause malformation and curling of the petals and also may cause side effects such as fruit hollowness, quality reduction and the like. According to the scheme, the gene technology is ingeniously utilized to directly act on a plant endogenous regulation and control system, the synergistic effect of strawberry petal curling and quality optimization is achieved by activating or inhibiting specific gene expression (such as cell division and nutrient distribution), and in addition, the over-expression FvePREs transcription factor can be used for improving the yield of strawberry petals by changing the hereditary characteristics of strawberries. The good petal curling effect can be kept in the long-term planting process, dependence on external chemical substances can be reduced, and sustainable agricultural development is facilitated.

The invention relates to the field of gene technology application, in particular to a tomato salt stress gene SlGLUB, a primer and application of the tomato salt stress gene SlGLUB. In the invention, the SlGLUB gene can promote the root development of the tomato under normal conditions, and after salt stress treatment, the SlGLUB gene can improve the salt tolerance of the tomato by adjusting the root development of the tomato plant and increasing the total root length and the root surface area, and from the result of paraffin sections of roots, stems and leaves, the salt tolerance of the tomato can be improved on the cellular level. The SlGLUB gene can reduce the damage of salt stress to tomato plants, the SlGLUB gene can improve the capability of removing active oxygen by improving the activity of antioxidant enzyme, and the membrane lipid peroxidation degree can be reduced, so that the salt resistance of the tomato plants is improved.

The invention relates to the technical field of biological genes, in particular to a gene directed editing system applied to mycoplasma bovis, which is an editing system for expressing a combination of a single plasmid of GP35 single-chain annealing protein and a targeting fragment ssDNA (single-stranded deoxyribonucleic acid), and comprises an expression plasmid of a GP35 gene and the ssDNA of the targeting fragment. The invention relates to a gene directed editing method applied to mycoplasma bovis. The gene directed editing method comprises the following steps: constructing a mycoplasma bovis strain for expressing GP35 single-chain annealed protein, constructing an ssDNA single chain of a targeting fragment, and performing gene editing on a target gene to obtain a pdhC gene deleted strain. According to the invention, a KanR gene is successfully inserted at the 158bp position of a pdhC gene coding region of a mycoplasma bovis PG45 strain, so that a mycoplasma bovis pdhC gene oriented deletion strain is obtained. According to the invention, the metabolism-related gene directed deletion strain M. bovis PG45 [delta] pdhC is successfully constructed, and the pdhC protein is characterized to have the ability of regulating the growth rate of mycoplasma bovis and maintaining colonial morphology.

The invention discloses a multi-constraint-based anti-parallel DNA triplex design method and system, and relates to the technical field of DNA triplex targeting genes, and the method comprises the following steps: constructing an mCGR initialization model, adjusting a matrix iteration rule according to a sequence length and a basic constraint, and generating all possible sequence sets satisfying the constraint; mapping the sequence set into flow network nodes, screening out approximate nodes through similarity search, constructing to-be-screened edges, then performing sequence comparison on the to-be-screened edges, only reserving the edges of the sequence within a preset stem length range, and deleting the edges beyond the range, so as to overcome the problem of high complexity of mCGR during combined constraint calculation; and finally, outputting a DNA triplex coding sequence meeting all constraint conditions. According to the method, the anti-parallel DNA triplex TFO sequence meeting multiple constraints can be efficiently generated, the sequence set scale is large, the stability is high, the crosstalk rate is low, and a key technical support is provided for research and development of nucleic acid drugs in the field of targeted gene therapy.

This invention relates to the field of gene technology, and more particularly to the preparation of a zebrafish model lacking Trmt61a enzyme activity, for the purpose of preparing a stable genetic model. trmt61a D181A -P2A-EGFP Knock-in zebrafish strains. This invention utilizes CRISPR / Cas9 technology to... trmt61a d181a trmt61a d181a The mutation site was specifically introduced into the zebrafish genome, resulting in the loss of Trmt61a catalytic activity and tRNA m 1 A zebrafish model with significantly reduced tRNA modification levels. This zebrafish model fills a research gap and provides a basis for tRNA m... 1 A provides a usable in vivo research model for studying the biological developmental regulatory mechanisms related to epigenetic modifications and elucidating the pathological mechanisms of diseases (such as malignant tumors and hematopoietic dysfunction), which can be directly used to explore the core role of this epigenetic modification in development and disease.

The invention belongs to the technical field of genes, discloses a method for detecting and mining associated genes and regulatory pathways by using double-layer optimization, solves the black box problem in a machine learning prediction process and provides biological mechanism explanation. The previous research mostly utilizes multi-omics data to predict phenotypes, but a machine learning mode is difficult to really evaluate the action size and action approach of genes, and lacks interpretability at the same time. According to the RADAR, a parameter is given to each omics of each gene for double-layer optimization solution, phenotypes can be explained to the maximum extent, and meanwhile the action degree of each omics can be evaluated. In addition, RADAR can process input data of any number of omics, and a program can automatically identify the number of the omics and calculate the effect size of each omics. RADAR can accept omics input of different gene numbers, and the actual data condition is better met.

This invention belongs to the field of gene technology and discloses a method for efficient expression of exogenous proteins based on the NC_048601.1 site within the CHO cell genome. The site within the CHO cell genome used for stable protein expression is located within the range of bases 16027000-16033000 of NC_048601.1 in the CHO cell genome, and its nucleotide sequence is shown in SEQ ID NO: 1. This invention introduces different protein genes into a fixed location within the CHO cell genome and achieves stable expression.

The invention relates to a gynostemma pentaphylla oxidized squalene cyclase gene GpOSC2 and application thereof, and belongs to the technical field of genes. The amino acid sequence of the gynostemma pentaphylla oxidized squalene cyclase GpOSC2 is as shown in SEQ ID NO. 1. A coding sequence for coding the gynostemma pentaphylla oxidized squalene cyclase GpOSC2 gene is a nucleotide sequence as shown in SEQ ID NO. 2. According to the present invention, the gene is adopted to carry out fermentation so as to obtain Dammarenediol-II, lupeol, lanosterol, cycloartenol and alpha-amyrin alcohol, such that the application prospect is good, and the promotion and the application are easy.

The invention belongs to the technical field of optimized coding genes, and particularly relates to recombinant human elastin and a preparation method and application thereof.The amino acid sequence of the recombinant human elastin is shown as SEQ ID No.1. Experiments prove that the protein is free of cytotoxicity, high in safety and good in cell proliferation and migration promoting effect, and the recombinant human elastin can be used for preparing the recombinant human elastin. The method can be widely applied to the fields of medicines, medical instruments, biological materials, tissue engineering and the like.

The invention discloses a dsRNA fragment capable of improving wheat amylopectin content and a synthesis method and application thereof, and relates to the field of wheat starch content regulation, pET28a is used as an initial skeleton, a sequence containing a reverse T7 promoter, multiple cloning sites and ISA1 or ISA2 gene fragment is inserted, a recombinant vector is constructed through restriction enzyme digestion connection of restriction enzymes SalI and EcoRI, and the recombinant vector is used as a recombinant vector for improving wheat amylopectin content. And transforming the recombinant expression vector into an escherichia coli HT115 competent cell, and synthesizing the dsRNA fragment after induced expression. According to the invention, a non-transgenic transient silencing technology is adopted, and target gene expression is inhibited in a targeted manner, so that the problems of long period, low efficiency, dependence on complex gene cloning and vector construction processes and potential off-target effect and ecological risk existing in the traditional gene technology are avoided.

The invention discloses a construction method and application of an hMRGPRX3 humanized mouse model and an atopic dermatitis model, and relates to the technical field of disease model construction. The whole gene sequence of the human MRGPRX3 or a vector containing the gene sequence of the hMRGPRX3 is transferred into mouse genome DNA (Deoxyribonucleic Acid) by utilizing a transgenic technology, and a functional human hMRGPRX3 receptor can be simulated and expressed in a mouse model more truly by transferring upstream and downstream regulatory sequences of the hMRGPRX3 gene. The pruritus signal transduction mediated by the receptor in human dorsal root ganglion sensory neurons can be accurately simulated, and signal deviation caused by species difference of a traditional mouse model is overcome.

The invention discloses a method for screening a transcriptional regulatory element and application thereof, relates to the technical field of genes, and provides a method for predicting a collaborative transcriptional regulatory element based on combination of single-cell multi-omics and experimental exploration. The potential synergistic effect between transcriptional regulation elements, chromatin remodeling complex interaction and other cross-level regulation events can be revealed, and a method is provided for recognition of disease key regulation nodes. On the other hand, the application systematically reveals key transcriptional regulatory elements involved in disease responses based on single cell multi-omics sequencing techniques. Through integration of single cell chromatin accessibility analysis and transcriptome sequencing data, interaction among a core transcription factor, an epigenetic regulation element and a signal transduction pathway for driving disease generation is clarified, and the mononuclear cell specific epigenetic-transcriptional regulation network is successfully constructed.

The application belongs to the technical field of genes, and discloses a method for efficiently expressing exogenous proteins based on a NC_048604-1 site in a CHO cell genome. The site for stably expressing proteins in the CHO cell genome is located in a 94142000-94148000 base range of the NC_048604-1 in the CHO cell genome, and the nucleotide sequence is shown as SEQ ID NO:1. In the fixed position in the CHO cell genome, different protein genes are introduced and stably expressed.

The invention discloses an FvePREs transcription factor and application thereof in promoting strawberry fruit expansion, compared with a traditional expansion agent, the scheme has the advantages that the safety is improved, and the existing expansion agent may cause malformed fruits and hollow fruits and has chemical residue risk. The FvePREs transcription factor regulates and controls fruit development through a molecular level, exogenous hormones are not needed, and chemical pollution and potential food safety hazards are avoided; the scheme is more accurate in action mechanism, the existing biological agent (such as a formula containing alginic acid and brassinolide) needs to depend on external application, and the effect is influenced by environmental factors. The gene technology directly acts on a plant endogenous regulation and control system, and the synergistic effect of fruit expansion and quality optimization is achieved by activating or inhibiting specific gene expression; the scheme has sustainability, the overexpression FvePREs transcription factor can change the genetic characteristics of strawberries, so that the strawberries can keep a good expansion effect in the long-term planting process, dependence on external chemical substances is reduced, and sustainable agricultural development is facilitated.

The present invention discloses the use of a biological factor Sparcl1 in a drug that promotes the regeneration of inner ear hair cells, belonging to the field of gene technology for inner ear cell regeneration. The biological factor Sparcl1 is used to promote the regeneration of inner ear stem cells into hair cells. The amino acid sequence of the biological factor Sparcl1 is shown in SEQ NO.1. Compared with the prior art, this application explores the regenerative effect of the Sparcl1 gene on inner ear hair cells by combining gene regulation with the restoration of auditory function by inner ear stem cells. This patent uses AAV-ie as a viral vector for delivering the target gene, and then injects it through the round window of the ear of a newborn mouse. This method has the highest delivery efficiency and causes less damage to the cochlea, fundamentally avoiding the problems of low vector transfection efficiency and the choice of delivery method. Secondly, this patent uses human recombinant Sparcl1 protein, which can act directly on humans and therefore has important value and role in the regeneration of inner ear hair cells and the recovery of hearing loss in clinical practice.

The application belongs to the technical field of genes, and discloses a method for efficiently expressing exogenous proteins based on a Dlg3 gene site in a CHO cell genome. The Dlg3 gene site for stably expressing proteins in the CHO cell genome is located in the range of 69498000-69504000 bases of the CHO cell genome NC_048604.1, and the nucleotide sequence is shown as SEQ ID NO:1. In the fixed position of the CHO cell genome, different protein genes are introduced and stably expressed.

This invention relates to the field of gene technology, specifically disclosing a method for enhancing the secretion of secondary metabolites based on the regulation of quorum sensing signal molecules. This method activates the expression of bacterial secondary metabolite synthesis gene clusters through bacterial-fungal interaction, comprising the following steps: (a) co-culturing bacteria and fungi in contact or non-contact mode; (b) during co-culturing, the fungi secrete the quorum sensing signal molecule farnesol; (c) the bacteria sense farnesol through a two-component system, activating the expression of secondary metabolite synthesis genes. This invention effectively overcomes the gene silencing problem caused by metabolic load or lack of regulation in natural strains by activating the originally trace expression of secondary metabolite synthesis gene clusters in bacteria using the fungal-derived quorum sensing signal molecule farnesol.