22063results about "Plant genotype modification" patented technology

The invention discloses an AsMYB19 gene and application thereof, and relates to the technical field of biology, the nucleotide sequence of the AsMYB19 gene is shown as SEQ ID NO.1, the AsMYB19 gene can regulate and control the tiller number and the yield of oat, gene resources and strategies are provided for oat breeding and improvement of oat germplasm resources, the breeding speed is increased, and the breeding efficiency is improved.

The invention discloses a biosynthesis system for high-yield 1, 6-hexamethylenediamine and application of the biosynthesis system, and belongs to the technical field of biology. According to the method, key genes which are excavated and optimized in a 1, 6-hexamethylenediamine biosynthetic pathway are co-expressed in a single cell module or a multi-cell module, and different enzyme elements expressed by the cell module are subjected to enzyme cascade reaction, so that 1, 6-hexamethylenediamine can be efficiently synthesized by taking caprolactam or 6-aminocaproic acid as a substrate; furthermore, after the carboxylic acid reductase MaCAR is mutated, the catalytic activity of the carboxylic acid reductase MaCAR can be remarkably improved, and the yield of the 1, 6-hexamethylenediamine is further improved; in addition, the biosynthesis system disclosed by the invention directly takes the cell module as a whole-cell catalyst, and an enzyme purification step is avoided, so that the biosynthesis system has the advantage of low cost.

The invention discloses an AsSN1 gene and application, and relates to the technical field of biology. The nucleotide sequence of the AsSN1 gene is as shown in SEQ ID NO.1, and the amino acid sequence of the AsSN1 protein is as shown in SEQ ID NO.2. The AsSN1 gene and the protein provided by the invention can regulate and control the spikelet number of the oats, further regulate and control the yield of the oats, provide important support for oat breeding and germplasm resource improvement, and have important significance for meeting the development requirements of animal husbandry.

The invention discloses a glycosyl transferase mutant and an application of the glycosyl transferase mutant in synthesis of rebaudioside. According to the invention, single-point mutation and multi-point mutation are carried out on the basis of a glycosyl transferase amino acid sequence as shown in SEQ ID NO: 1, and a mutant with improved catalytic activity is obtained. The catalytic activity, the substrate specificity and / or the substrate specificity of the glycosyl transferase mutant are / is changed, and the catalytic activity of enzyme to a specific substrate can be remarkably improved by mutation at a specific site. And carrying out induced expression and protein purification on the obtained mutation sequence to obtain the mutant enzyme. The mutant enzyme is used as a catalyst, and UDPG is used as a glycosyl donor, so that the catalytic reaction efficiency of substrates such as stevioside ST, rebaudioside A (RebA) and rebaudioside D (RebD) can be obviously improved. According to the glycosyl transferase UGT76G1 mutant constructed by the research, the catalytic activity of the glycosyl transferase UGT76G1 mutant is improved, and efficient production of rebaudioside M is realized by optimizing a reaction system.

The invention relates to the technical field of genetic engineering, and particularly discloses a genetic engineering bacterium for producing melanin as well as a construction method and application of the genetic engineering bacterium. According to the application, the yield of the melanin can be effectively increased by modifying a metabolic pathway of the genetically engineered bacterium, and the yield of the melanin reaches 6.2 g / L and 18.5 g / L respectively through shake-flask culture and 5L fermentation tank culture. Compared with the prior art, the yield of melanin produced by the genetically engineered bacterium is improved by 30% at the shake flask fermentation level, the fermentation yield of a fermentation tank is improved by 4.3 times, in addition, expensive tyrosine is changed into glucose as a fermentation precursor substance, the production cost is greatly reduced, and an efficient and feasible solution is provided for large-scale production of melanin.

The invention discloses a bovine I-type alpha interferon-ferritin fusion protein, a mutant thereof, a preparation method and an application of the bovine I-type alpha interferon-ferritin fusion protein. The bovine I-type alpha interferon is fused with a ferritin subunit, and interferon molecules are highly repeatedly and orderly displayed on the surface of a ferritin nanocage by utilizing the self-assembly characteristic of ferritin, so that the expression level, the structural stability and the antiviral activity of the interferon are remarkably improved. The fusion protein is further subjected to single-site or multi-site rational design mutation, and a mutant with significantly improved antiviral activity and stability is obtained. According to the invention, a silkworm or insect cell eukaryotic expression system is adopted to express the fusion protein or the mutant thereof, and the expression system is safe to operate, simple and convenient in procedure, low in cost and extremely beneficial to large-scale industrial production; the prepared fusion protein or mutant nanoparticles have application prospects in preparation of drugs or reagents for preventing or treating bovine viral diseases.

A Chinese cabbage hybrid seed production marking method relates to the technical field of Chinese cabbage hybrid seed production. The technical scheme of the method is as follows: after Chinese cabbage hybrid seed production pollination is finished, firstly holding the pedicel of a pollinated flower with the left hand; then transversely tearing off two sepals from the root with the right hand, wherein light movement is required. The Chinese cabbage hybrid seed production marking method can perform marking without rope tying, ring arranging or stamp-pad ink coating, and is low in cost and high in accuracy.

The invention provides a production strain of an antibiotic-free plasmid, the production strain is a gene editing strain of a PIR strain and is named as PIR1-WN:: 0636 or PIR1-PR: 0636, the production strain contains a nucleotide sequence for coding toxin protein and the antibiotic-free plasmid, and the antibiotic-free plasmid contains a nucleotide sequence for coding antitoxin protein; and preferably, the replicon DNA element of the nonreactive plasmid is R6K-gamma. The toxin protein gene of the production strain disclosed by the invention can be stably passaged, has lethality after being induced and can be used for plasmid screening; according to the invention, the positive rate of transforming the nonreactive plasmid into the PIR1-WN:: 0636 strain is more than 80%, and stable production of the plasmid with a high superhelix ratio can be realized.

The invention discloses a rational copolymerization strategy for improving the stability of an oligomeric structure of acid-induced high-order oligomeric decarboxylase AdiA in a neutral to alkaline environment and application of the rational copolymerization strategy, and belongs to the field of bioengineering. According to the strategy, rational charge overturning transformation is carried out on a pentamer radial oligomeric interface and a dimer weft-wise oligomeric interface at the same time, and stable assembly and efficient catalysis of a decamer under the condition that the pH value is from 7.0 to 9.0 from neutral to alkaline are achieved. Wherein positive charges are introduced into a meridian interface to weaken electrostatic repulsion, and negative charges are introduced into a latitudinal interface to enhance dimer compactness and substrate transfer efficiency. The specific enzyme activity of the representative double mutant AdiAD471K / E467K / H736E is 45.5 times that of a wild type when the pH value is 8.0, and the representative double mutant AdiAD471K / E467K / H736E keeps a complete decamer state when the pH value is 7.0-9.0. The yield of butanediamine is up to 156.5 g / L by using the mutant to catalyze whole cells.

Provided are S-adenosylmethionine synthetase mutants and the use thereof, belonging to the technical field of biological enzyme engineering. Site-directed combinatorial mutations are performed on positions 189, 200, 234, 266 and 371 of S-adenosylmethionine synthetase MAT to obtain a series of mutants with improved enzyme activity, wherein the specific enzyme activity of I189V / V266H and V266H / N371R is increased by more than 10 times relative to the parent. The yields of S-adenosylmethionine synthesized by whole cells of recombinant bacteria expressing mutants I189V / V266H / N371R, V266H / Q234N / N371R and I189V / V266H / Q234N / N371R all reach 1550 mg / L or more. The provided enzyme mutants have broad application prospects in the production of S-adenosylmethionine and the construction of genetically engineered microorganisms for S-adenosylmethionine.

The invention relates to the technical field of biology, and particularly discloses a recombinant collagen III and application thereof in preparation of gel. The recombinant collagen III is designed by optimizing functional area sequences of human I-type and III-type collagen, the amino acid sequence is shown as SEQ ID No.2, and the recombinant collagen III has the characteristics of high stability, good hydrophilicity and low immunogenicity. The preparation method comprises the steps of expression vector construction, escherichia coli induced expression, affinity chromatography purification and renaturation. The recombinant collagen III can be prepared into a gel dressing and comprises sodium alginate, methylparaben and other components. Experiments show that the gel can effectively promote cell proliferation and has no cytotoxicity; in a mouse skin injury model, the collagen can relieve inflammation, accelerate wound healing and inhibit scar formation, and the effect of the collagen is superior to that of natural human III-type collagen. The invention provides a safe and efficient novel material for wound repair, and is suitable for medical dressings and tissue engineering.

The invention discloses a recombinant XVII type collagen engineering bacterium as well as a preparation method and application thereof, and belongs to the technical field of genetic engineering, and the recombinant XVII type collagen engineering bacterium is realized through three steps of designing and optimizing collagen, improving translation through codon optimization and selecting a vector for cloning. By optimizing the amino acid sequence and the codon, the expression efficiency and the stability of the recombinant XVII type collagen in engineering bacteria are improved. The optimization not only improves the production efficiency of the protein, but also reduces the production cost. The recombinant XVII type collagen has potential in the medical field, especially in the fields of skin repair, wound healing, aging resistance and the like. Meanwhile, in the fields of beauty and biological materials, collagen is widely applied to products such as masks and skin care products as a basic component, and can promote skin cell regeneration and enhance skin elasticity and repair capacity.

The invention relates to a construction method of escherichia coli engineering bacteria with high yield of N-acetylglucosamine. According to the method, the metabolic network of escherichia coli is directionally modified through metabolic engineering, the problems of'product re-decomposition ', by-product accumulation, poor genetic stability and the like of natural strains are solved, and efficient industrial production of GlcNAc is realized.

The invention relates to the technical field of biological catalysis, and discloses an L379A mutant enzyme for preparing rebaudioside I. The enzyme is obtained by the following mutations generated by UGT76G1: leucine of the 379th amino acid sequence is mutated into alanine; the enzyme can be applied to RA in-vitro conversion preparation of RI with higher utilization value, the conversion rate is higher than 50%, and the enzyme activity is remarkably improved by 7 times compared with the original enzyme catalytic conversion enzyme activity. The UGT76G1 mutant disclosed by the invention has the advantages that (1) the blank is filled, and a special enzyme catalyst capable of realizing efficient in-vitro synthesis of rebaudioside I (RI) is provided for the first time; 2) efficiency jump: the catalytic conversion rate is greatly increased from original about 7% to more than 50% (increase gt; and 3) stability and reliability: in the cross-scale reaction of 10mL to 5L, the catalyst has stable catalytic performance, shows excellent industrial application potential, and is suitable for popularization and application.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to an SCRE10 gene for improving rice disease resistance and application of the SCRE10 gene, the base sequence of the SCRE10 gene is shown as SEQ ID NO.1, and the amino acid sequence of the SCRE10 gene is shown as SEQ ID NO.2. Through construction of a dexamethasone induced expression SCRE10 transgenic rice plant, it is found that the transgenic rice can significantly induce PR gene expression and active oxygen outbreak, and the resistance of rice to false smut and bacterial leaf blight can be improved. The invention proves that heterologous inducible expression of the Ustilaginoidea virens SCRE10 gene has the function of positively regulating the disease resistance of the rice, and the SCRE10 gene can be used for improving the disease resistance of the rice, which is of great significance to the creation of disease-resistant germplasm of the rice.

The present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to human cMET. The disclosure also provides polynucleotides encoding the antibodies or antigen binding fragments thereof, vectors comprising the polynucleotides, host cells comprising the vectors, methods of generating the antibodies, and compositions comprising the antibodies.

This invention provides a humanized monoclonal antibody that binds to advanced glycation end product (AGE) modified proteins or peptides on a cellular surface. [Solution] A humanized monoclonal antibody for advanced glycation end products, comprising at least one amino acid sequence selected from a group consisting of eight specific amino acid sequences, each with a different sequence. The antibody may be bound to a carboxymethyllysine-modified protein or peptide on a cell. The antibody can be used in a cell separation process, for example, in magnetic cell separation.

The invention belongs to the technical field of biological dyes, and discloses a bio-based melanin precursor compound, bio-based melanin as well as a preparation method and application of the bio-based melanin precursor compound and the bio-based melanin. The invention provides a bio-based melanin precursor compound 4-[5-(2, 2-diaminoethyl)-2, 3-dihydroxyphenyl]-1H-indole-5, 6-diphenol, and a preparation method of the bio-based melanin precursor compound. The bio-based melanin precursor compound is stable in chemical property and easy to store. And the catalyst can be used as a raw material to efficiently synthesize bio-based melanin through a catalytic oxidation reaction. The process for preparing the bio-based melanin is simple, mild, high in yield and suitable for large-scale industrial production. The bio-based melanin prepared by the invention has good application performance during dyeing, does not generate harmful substances, and is safe and environment-friendly.

The invention discloses application of a Chinese rose gene RhMYB1 in regulation of Chinese rose axillary bud germination. Relates to the technical field of molecular biology, and provides application of a product for inhibiting expression of an RhMYB1 gene or transcriptional translation protein of the RhMYB1 gene in regulation of axillary bud germination of Chinese roses, and a nucleotide sequence of the RhMYB1 gene is shown as SEQ ID NO.1. According to expression results of the RhMYB1 gene in buds at different sites of Chinese roses, it is found that the expression quantity of the RhMYB1 gene in upper buds (namely active buds) is high. The RhMYB1 gene in the axillary bud is instantaneously silenced, the silent RhMYB1 gene is found to significantly inhibit the germination and growth of the axillary bud, and the overexpression RhMYB1 can promote the growth of the axillary bud.

The invention discloses a high-stability hydroxysteroid dehydrogenase mutant, a co-immobilized enzyme construction method and application. According to the invention, a site-directed mutant library is constructed based on computer-aided design, and mutants with significantly improved thermal stability and greatly improved catalytic efficiency compared with wild type mutants are obtained through screening; meanwhile, a co-immobilized enzyme construction method based on electrostatic adsorption and chemical crosslinking is established, the hydroxysteroid dehydrogenase mutant and lactic dehydrogenase are co-immobilized, the co-immobilized enzyme can still keep high activity under high substrate concentration, and the conversion rate is still maintained to be 90% or above after 30 batches of co-immobilized enzyme are repeatedly used. And an important foundation is laid for industrial application of the hydroxysteroid dehydrogenase.

The invention relates to a CHO-S cell strain capable of stably expressing H5N1 hemagglutinin protein and a construction method of the CHO-S cell strain, and belongs to the field of bioengineering.H5N1 hemagglutinin protein expression plasmids are obtained in an in-vitro synthesis and seamless cloning mode, 293T cells are transfected through the H5N1 hemagglutinin protein expression plasmids, and it is proved that the H5N1 hemagglutinin protein is expressed; the CHO-S cell strain capable of stably expressing the H5N1 hemagglutinin protein is obtained by transfecting CHO-S cells by using the H5N1 hemagglutinin protein expression plasmids and carrying out multiple rounds of cloning and screening, so that a basis is provided for obtaining H5N1 recombinant protein influenza vaccines.

The invention belongs to the technical field of gene engineering, and particularly relates to a mutant of polymyxin efflux transporter and application of the mutant. The mutant is a PmxD transporter mutant, the amino acid sequence of the PmxD transporter mutant is shown as SEQ ID NO: 1, and compared with wild type PmxD, the polymyxin transport capacity of the PmxD transporter mutant (T38W) is improved by 450.46%; and the total discharge amount of polymyxin is increased by 85.72%. Meanwhile, the mutant can significantly improve the growth ability of the strain on a plate containing 250 [mu] g / mL of polymyxin B, namely significantly improve the autoresistance of paenibacillus polymyxa to polymyxin.

The invention discloses an inositol dehydrogenase mutant and a preparation method of D-chiral inositol, and belongs to the technical field of genetic engineering. The inositol dehydrogenase mutant is obtained by mutating an amino acid sequence as shown in SEQ ID NO.2 through the following mutations: the 200th amino acid is mutated into C from V, the 234th amino acid is mutated into Q from V, and the 256th amino acid is mutated into E from R. According to the invention, the amino acid sequence of wild type inositol dehydrogenase is mutated to obtain the inositol dehydrogenase mutant which is a high-activity mutant capable of catalyzing conversion of myo-inositol into 2-keto-myo-inositol; therefore, when the inositol dehydrogenase mutant and keto isomerase act together to prepare D-chiral inositol by taking myo-inositol as a substrate, the yield of the D-chiral inositol is effectively improved.

The invention discloses protease for improving the resistance of potatoes to late blight as well as a coding gene and application thereof, and belongs to the technical field of genetic engineering and plant disease resistance breeding. The amino acid sequence of the protease StRD21 is as shown in SEQ ID NO.2, or the protease StRD21 has a derivative sequence with the same disease-resistant function. The nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1. The invention also provides a specific primer pair for cloning the gene, a potato recombinant overexpression vector containing the gene and a transgenic host cell. The StRD21 gene is introduced into potatoes and overexpressed, so that the resistance of the potatoes to late blight can be remarkably enhanced. The invention provides an effective gene resource and a biotechnological means for cultivating a new variety of disease-resistant potatoes.

The application provides a wheat flour wet gluten content main effect site and SNP marker and application, and belongs to the wheat breeding technical field, and mainly provides SNP molecular markers Marker45889, Marker45896, Marker45348, Marker45403, Marker45397 and Marker45431, and the SNP site provided by the application has important significance for detecting and breeding wheat flour high FWG varieties, lines and breeding materials, and molecular markers can be developed according to the SNP site, so that the wheat quality breeding efficiency is accelerated.

The invention discloses a 3-sterone-delta1-dehydrogenase mutant and application of the 3-sterone-delta1-dehydrogenase mutant in preparation of a steroid drug intermediate. The mutant is obtained by performing single mutation or multiple mutation on 551, 468, 48 and 157 sites of an amino acid sequence of 3-sterone-delta1-dehydrogenase as shown in SEQ ID NO: 1. The 3-ketosterone-delta1-dehydrogenase is subjected to molecular modification through directed evolution and semi-rational design, the mutant with improved enzyme activity is screened in a high-throughput mode, the mutant can effectively improve the conversion rate of a substrate 11alpha, 17alpha-dihydroxyprogesterone, and the substrate conversion rate of the mutant ISM-2 is improved to 92.4% compared with 36.7% before mutation.

The invention belongs to the technical field of microbial genetic engineering, and particularly relates to an alpha-2, 3-sialyltransferase mutant and application thereof. The a-2, 3-sialyltransferase mutants V175N, P198A, G173D, D174Q, F196K and A315H are obtained by performing site-specific mutagenesis on six amino acids in an a-2, 3-sialyltransferase amino acid sequence coded by a gene derived from Campylobacter jejuni (OH4384), the yield of 3 '-sialyllactose is remarkably increased by single-point mutagenesis of the V175N, the yield of 3'-sialyllactose is remarkably increased by single-point mutagenesis of the V175N, the yield of 3 '-sialyllactose is remarkably increased by single-point mutagenesis of the P198A, the G173D, the D174Q, the F196K and the A315H, the optimal recombinant microbial strain 3 '-SL-09 (V175N / P198A / D174Q) is obtained through combination of multiple mutation sites, the yield can reach 5.91 g / L, the bottleneck problem of low catalytic efficiency of sialyltransferase is effectively solved, and an enzyme element with excellent performance is provided for high-efficiency and low-cost biological manufacturing of 3'-SL.

Provided in the present invention are a Brassica napus drought-tolerant gene BnaC04ARL and the use thereof. The gene is expressed in a plant to improve drought tolerance of the plant. The gene comprises a nucleotide sequence selected from the group consisting of: a nucleotide sequence shown as SEQ ID NO: 1; and / or a nucleotide sequence encoding an amino acid sequence shown as SEQ ID NO: 2. The overexpression of the gene can improve the viability of Brassica napus under drought conditions, enhance root system development, and improve the thousand-grain weight, the grain yield per plant, the number of siliques per plant, dry weight per plant, silique length, etc.

The invention discloses an aspergillus niger recombinant bacterium AnCT-XynB-K-agdA and application thereof, and belongs to the technical field of microbiology and bioengineering. According to the invention, Aspergillus niger AnCat is taken as an expression host, firstly, an acid protease regulatory factor prtT is knocked out to obtain a defective strain, and then different chaperonins are subjected to fusion expression by optimizing integration sites of a laccase lcc9 expression cassette, so that the high-expression laccase chaperonin is obtained. According to the method, an expression vector pC3-5 'agdA-PcitA-XynB-L-lcc9-hph-3' agdA is constructed, a recombinant bacterium AnCT-XynB-K-agdA is further obtained, the enzyme activity of the extracellular laccase subjected to shake flask fermentation reaches 1821.2 U / L, and finally, the recombinant efficient expression of the laccase Lcc9 in aspergillus niger is realized.

To provide a pharmaceutical composition for treating a cancer patient having a tumor having a total passenger gene mutation amount larger than the background mutation amount of the tumor.SOLUTION: A pharmaceutical composition for treating a subject having a tumor with a total passenger gene mutation load that is greater than the background mutation load of the tumor, wherein the background mutation load has been determined based on randomly selected genes of the tumor, comprising antibodies that bind to PD1 as an active ingredient. Antibodies that bind PD1 comprise a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 21 and / or comprise a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 22.SELECTED DRAWING: Figure 1