52108results about "Plant genotype modification" patented technology

The invention provides an anti-human serum albumin nano antibody and application thereof, and belongs to the technical field of antibodies. The invention provides an anti-human serum albumin nano antibody HSA-32 with relatively high affinity, and the anti-human serum albumin nano antibody HSA-32 is subjected to humanized transformation, so that three obtained humanized anti-human serum albumin nano antibodies can be used for reducing the immunogenicity of the HSA-32 and improving the immunogenicity of the HSA-32; and high specificity and high affinity of combination with the human serum albumin and stability and solubility of the antibody are maintained. The anti-human serum albumin nano antibody can be used for separation, purification and detection of human serum albumin and separation and purification of polypeptide, protein or antibody drugs fused with the anti-human serum albumin nano antibody. Meanwhile, the anti-human serum albumin nano-antibody can effectively prolong the half-life period of polypeptide, protein and antibody drugs fused with the anti-human serum albumin nano-antibody, and can be used for developing long-acting polypeptide, protein or antibody drugs.

A recombinant humanized collagen and a preparation method therefor. The recombinant humanized type III collagen can be efficiently and stably expressed in three expression systems of yeast, Escherichia coli, and CHO cells, and has the biological activity superior to that of a commercially available collagen product. The recombinant humanized type III collagen has structural characteristics and functional activity similar to those of a natural collagen, and thus has wide application prospects in biomedical materials, tissue engineering products, cosmetics, foods, health care products or drugs, medical instruments, medical cosmetology and the like.

The present invention discloses a human serum albumin-targeting nanobody and its application. A variety of human serum albumin-targeting nanobodies that can be used to construct long-acting fusion protein drugs and their immune screening methods are disclosed. The antibody is composed only of the heavy chain variable region and can be fused and expressed with therapeutic proteins or polypeptide drugs such as interleukin, interferon, and tumor necrosis factor in an Escherichia coli expression system. The fusion protein has the activity of specifically binding to human serum albumin, has a relatively high affinity, can effectively extend the drug half-life, and does not affect the biological activity of the recombinant protein drug at the same time.

The invention belongs to the technical field of gene engineering, and particularly relates to a beta-glycosidase SS-BGL mutant for preparing rare ginsenosides and application of the beta-glycosidase SS-BGL mutant. The beta-glycosidase SS-BGL mutant is obtained by conducting single-site mutation or double-site mutation on beta-glycosidase SS-BGL with the amino acid sequence shown as SEQ ID NO.1. The beta-glycosidase SS-BGL mutant is obtained by conducting single-site mutation or double-site mutation on beta-glycosidase SS-BGL with the amino acid sequence shown as SEQ ID NO.2. Wherein the single site mutation is that phenylalanine at the 229 site of SEQ ID NO. 1 is mutated into proline; the double-site mutation is as follows: phenylalanine at the 229 site in SEQ ID NO. 1 is mutated into proline, and glycine at the 301 site is mutated into proline; or the phenylalanine at the 229 site in SEQ ID NO. 1 is mutated into proline, and the leucine at the 337 site is mutated into valine; or the phenylalanine at the 229 site in SEQ ID NO. 1 is mutated into the proline, and the glycine at the 338 site is mutated into the proline. On the basis of natural beta-glycosidase SS-BGL, the molecular structure of the beta-glycosidase SS-BGL is modified through site-directed mutagenesis, and four beta-glycosidase SS-BGL mutants with the saponin conversion activity improved are obtained.

The invention belongs to the technical field of cosmetics, and discloses anti-aging recombinant collagen as well as a preparation method and application thereof. The amino acid sequence of the anti-aging recombinant collagen is as shown in SEQ ID NO.1, and the nucleotide sequence of the anti-aging recombinant collagen is as shown in SEQ ID NO.2. The recombinant collagen CaI1 is constructed and expressed by recombining the effective sequences of the I-type collagen, the III-type collagen and the XVII-type collagen, and the recombinant collagen CaI1 is free of cytotoxicity, free of hidden danger of endotoxin and good in safety. The skin care product added with the recombinant collagen CaI1 can promote skin collagen absorption, inhibit skin aging and improve skin elasticity, and has a good application prospect.

The invention discloses a bovine serum albumin combined nano antibody and a preparation method and application thereof, the amino acid sequence of the bovine serum albumin combined nano antibody Nb1 is shown as SEQ.ID No.1, the nano antibody Nb not only has high affinity to bovine serum albumin, but also has good binding activity to both human serum albumin and mouse serum albumin, and the nano antibody Nb has high affinity to human serum albumin and mouse serum albumin. And the antibody has a relatively good broad-spectrum application range, and can be widely applied to the aspects of drug half-life period enhancement, tumor targeted therapy and imaging, immunodetection or diagnosis, recombinant albumin purification or enrichment and the like.

The invention belongs to the technical field of biology, and relates to an escherichia coli engineering strain for producing L-isoleucine through a methyl malic acid way as well as a construction method and application of the escherichia coli engineering strain. According to the invention, Escherichia coli BW25113 is used as a chassis strain, and a methyl malic acid synthase high-activity mutant gene cimA3.7, an isopropyl malic acid isomerase gene GsleuCD, a 3-isopropyl malic acid dehydrogenase gene AfleuB and a leucine dehydrogenase gene Lsleudh are over-expressed; a methyl malic acid absorption protein gene is over-expressed, and a branched chain amino acid transport system is modified to improve the exosome of the L-isoleucine; all methyl malic acid pathway related genes are integrated to a genome of a chassis strain, and a plasmid-free L-isoleucine production strain is constructed; the copy number of a key gene on a genome is optimized, and the yield of the engineering strain L-isoleucine is increased by introducing a non-oxidative glycolysis approach. The yield of the L-isoleucine after the engineering strain is fermented for 34 hours in a fermentation tank is 56.6 g / L.

To provide antibodies that bind to lymphocyte activation gene 3 (LAG3) protein, which is a T cell co-inhibitor, and methods of use.SOLUTION: In various embodiments of the invention, the antibodies are fully human antibodies that specifically bind to LAG3. In some embodiments, the antibodies of the invention are useful for inhibiting or neutralizing LAG3 activity, thus providing means of treating a disease or disorder such as cancer or viral infection. The antibodies of the present invention are useful, inter alia, for targeting immune cells expressing LAG3 and for modulating LAG3 activity.SELECTED DRAWING: None

The invention relates to the technical field of biology, in particular to a verticillium dahliae virulence gene, a verticillium dahliae virulence protein and application. The invention discloses a verticillium dahliae virulence gene VdPHO23like. The verticillium dahliae virulence gene VdPHO23like comprises a polynucleotide sequence for coding an amino acid sequence of SEQ ID NO: 3. According to the invention, expression of the gene is inhibited in a targeted manner through an RNA interference mediated gene silencing technology so as to weaken virulence of pathogenic bacteria, and an efficient, specific and environment-friendly comprehensive prevention and control strategy is provided for verticillium wilt of crops such as cotton.

The invention belongs to the technical field of gene engineering, and particularly relates to applications of OpgG and a mutant thereof in production of human milk oligosaccharide. The invention provides a dextran biosynthetic protein OpgG mutant, and the OpgG mutant is obtained by mutating the 443rd amino acid from threonine to proline on the basis of a wild type dextran biosynthetic protein OpgG as shown in SEQ ID NO.1. The invention also provides a preparation method of the dextran biosynthetic protein OpgG mutant. An opgG gene and / or a T443P mutant coding gene are / is introduced into a human milk oligosaccharide production strain and applied to production of human milk oligosaccharide, and the yield of 3 '-SL, 6'-SL, LNT II, LNnT and LNT is greatly increased.

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 present invention relates to the fields of genetic engineering and plant genetic breeding, and specifically to the application of wheat TaPND gene. In the present invention, by taking gene TaPND as a target, designing a sgRNA sequence based on CRISPR / Cas9, connecting a DNA fragment containing the encoding sgRNA sequence to a vector carrying CRISPR / Cas, a plant binary expression vector capable of knocking out the TaPND gene in wheat is constructed; by genetically transforming wheat, offspring with TaPND gene knockout are obtained, and wheat mutants with early flowering time are obtained. Experiments have confirmed that transgenic wheat with TaPND gene knockout achieves the advancement of wheat flowering time. The technical scheme of the present invention provides a practical method for realizing rapid wheat breeding using genetic engineering technology, which has important breeding application value and broad market application prospects.

The invention discloses a sugarcane germplasm resource evaluation and breeding method for smart agriculture. The sugarcane germplasm resource evaluation and breeding method comprises the following steps: step 1, collecting materials with wide genetic diversity; establishing an intelligent incubator and a greenhouse, and monitoring environmental parameters by using an Internet of Things sensor; 2, performing data acquisition on the related germplasm resources by using an unmanned aerial vehicle, a robot and a near infrared spectrum technology, and constructing a phenotype database for data management; meanwhile, genotype identification is carried out by combining molecular marker-assisted selection and genome selection technologies, molecular markers of key genes are mined, and the genetic value of related germplasm is evaluated; step 3, constructing a hybrid combination prediction model by using an AI algorithm, optimizing parent matching of the related germplasm and the cultivated sugarcane, monitoring and collecting the filial generation at the same time, and analyzing and screening in combination with smart agriculture; 4, demonstration planting is conducted on the bred excellent sugarcane strain, and application of the excellent strain is improved through government-enterprise-scientific research institution collaborative popularization and in combination with farmer technical training.

The invention provides a carbonyl reductase mutant. The carbonyl reductase mutant is obtained by performing single-point mutation or multi-point combined mutation on the 17th site, the 40th site and the 64th site of an amino acid sequence as shown in SEQ ID NO.1. The invention also provides a coding gene, a recombinant vector containing the coding gene, a co-expression engineering bacterium and an application of the co-expression engineering bacterium. Compared with a wild type enzyme, the carbonyl reductase mutant has high activity and high stereoselectivity, the enzyme activity can reach more than two times that of the wild type enzyme, the catalytic efficiency on a substrate precursor ketone is remarkably improved, the yield of a statin drug intermediate and chiral alcohol synthesized by an enzyme method of the carbonyl reductase mutant is remarkably improved, and the yield of a rosuvastatin intermediate (3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R) is remarkably improved. The highest yields of the (3R, 5R)-6-chloro-3, 5-dihydroxyhexanoic acid tert-butyl ester and the atorvastatin intermediate (3R, 5R)-6-cyano-3, 5-dihydroxyhexanoic acid tert-butyl ester reach 99% and 97.6% respectively, e.e. Is larger than 99%, and the method has high industrial application value.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody 4F6 for HPV16 type E7 protein detection and application. The heavy chain variable region CDR sequences and the light chain variable region CDR sequences of the antibody are SEQ ID NO.1-3 and SEQ ID NO.4-6 respectively, and the antibody can be specifically combined with HPV16 type E7 protein without cross reaction. A double-antibody sandwich ELISA detection system constructed based on the antibody is combined with a biotin-avidin amplification technology, the sensitivity reaches 100 pg / mL, and the antibody is suitable for rapid detection of the HPV16 type E7 protein in a cervical exfoliated cell sample. The invention further provides the HPV16 type E7 recombinant protein which is obtained through prokaryotic expression and has immunocompetence, and the HPV16 type E7 recombinant protein is applied to antibody screening and detection. Compared with nucleic acid detection, the method has a lower false positive rate, can be used for early screening of cervical cancer and precancerous lesions, and has a good application prospect.

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 provides humanized recombinant XVII collagen for promoting hair follicle stem cell repair and resisting aging as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The humanized recombinant XVII type collagen comprises polypeptide derived from the XVII type collagen or repeated protein formed by N times of repeated series connection of the polypeptide serving as a unit, n is an integer from 2 to 4; the polypeptide derived from the XVII type collagen comprises a polypeptide A and / or a polypeptide B; the amino acid sequence of the A polypeptide is as shown in SEQ ID NO: 1; and the amino acid sequence of the B polypeptide is as shown in SEQ ID NO: 2. The humanized recombinant XVII type collagen not only can be massively and stably recombined and expressed, but also has cell adhesion activity, cell proliferation promotion activity, cell migration promotion activity and excellent hair follicle repair and regeneration promotion activity, and has no sensitization side effect. The invention provides a new way for preparing collagen products with rich functions.

The invention relates to a recombinant A type humanized XVII type collagen as well as construction and application thereof, the protein is the humanized XVII type collagen which is composed of an intracellular region, a transmembrane region and an extracellular region, and the amino acid sequence is as shown in SEQ No. 1. The hair follicle repairing agent has the advantages of hair follicle repairing, hair regeneration, good biological activities of cell proliferation, cell adhesion and the like, and can be widely applied to the fields of medicines, medical instruments, biological materials, tissue engineering, cosmetics and the like. The gene sequence of the recombinant A type XVII collagen is 100% identical to the corresponding part of the gene sequence of the recombinant A type XVII collagen, so that the recombinant A type XVII collagen has good hydrophilicity and high activity, and has application potential in the fields of biomedical materials, beauty cosmetics, skin care products and the like.

The invention relates to the field of rice gene engineering, and particularly provides OsSULTR2; the invention also discloses application of the 2 protein and the coding gene thereof in regulating and controlling the salt tolerance of rice. The protein meets the following conditions: B1) a protein with an amino acid sequence of SEQ ID NO.1; and B2) a fusion protein with the same function obtained by connecting a tag to the N end and / or C end of B1). The method is used for detecting OsSULTR2; the salt stress phenotype identification in the seedling stage is carried out on the transgenic rice with the gene knockout 1, 2, and the result shows that when the gene segment is deleted, the salt stress tolerance of the rice is improved, and the function and the application way of the gene are proved. Therefore, the OsSULTR2 of the present invention; the 2 protein and the coding gene thereof can regulate and control the salt tolerance of rice, and have important significance for cultivating salt-tolerant transgenic rice.

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 high temperature resistant xanthan gum lyase mutant and a preparation method thereof, wild type xanthan gum lyase comes from Paenibacillus anensis, site-directed mutagenesis is carried out on the wild type xanthan gum lyase to obtain the high temperature resistant xanthan gum lyase mutant, and the mutant comprises at least one of N66V, R292A and S676V. The enzymatic activity of the xanthan gum lyase mutant treated in a buffer solution with the pH value of 5.0 at the temperature of 65-85 DEG C for 2 h is remarkably improved compared with that of an original enzyme, and the enzymatic activity of N66V / R292A / S676V is improved by 3.8 times compared with that of the original enzyme at the temperature of 85 DEG C.

The invention discloses a high-yield cold-resistant pennisetum germplasm screening method and application thereof, and the method is based on 10 yield traits, 4 cold-resistant indexes and 4 low-temperature photosynthetic ability indexes of pennisetum, constructs a multi-dimensional dynamic monitoring system across autumn, winter and spring, and performs high-yield cold-resistant comprehensive evaluation on the pennisetum germplasm, thereby screening out a high-yield cold-resistant breeding material. The invention provides a comprehensive, efficient and accurate high-yield cold-resistant Chinese pennisetum germplasm screening method, through a multi-dimensional dynamic monitoring system crossing autumn, winter and spring, yield traits, cold resistance indexes and low-temperature photosynthetic ability indexes are integrated, and the problems that a traditional method is single in evaluation dimension, disjointed with field reality and insufficient in dynamic adaptability are solved. The comprehensive evaluation system based on the principal component quantitative model not only fills the blank of the high-yield and cold-resistant evaluation model of the pennisetum alopecuroides, but also remarkably improves the breeding efficiency, provides powerful support for cultivating cold-resistant and yield-reduction-free breakthrough varieties, and has important application value and popularization prospect.

The invention relates to a saccharomyces cerevisiae strain for efficiently synthesizing cordycepin as well as a construction method and application of the saccharomyces cerevisiae strain, and belongs to the technical field of microbial genetic engineering. The construction method provided by the invention comprises the following steps: carrying out multi-copy genome integrated expression on heterologous codon optimized 2 '-carbonyl-3'-deoxyadenosine reductase genes and 3 '-adenosine monophosphate phosphohydrolase genes, so as to increase the dosage of key enzymes; and supplementing the defective gene of the engineering strain to obtain the saccharomyces cerevisiae strain B5U3 with high yield of cordycepin. According to the obtained saccharomyces cerevisiae strain for efficiently synthesizing the cordycepin, the cordycepin yield reaches 2.92 g / L after shake flask fermentation, the cordycepin yield reaches 7.5 g / L after fermentation in a fermentation tank, and a foundation is laid for safer and more reliable industrial production of the cordycepin.

The invention discloses preparation of a targeting LYPD3 antibody and application of the targeting LYPD3 antibody in tumor immunotherapy. The antibody or antigen binding fragment thereof comprises a heavy chain variable region having CDR-H1, CDR-H2 and CDR-H3 and a light chain variable region having CDR-L1, CDR-L2 and CDR-L3 wherein CDR-H1, CDR-H2 and CDR-H3 comprise the amino acid sequences of SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, respectively, and CDR-L1, CDR-L2 and CDR-L3 comprise the amino acid sequences of SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, respectively.

The invention discloses polyketide synthase or a polyketide synthase mutant and application thereof, and belongs to the technical field of gene engineering. The polyketide synthase is AspiPKS4, AspiPKS5, AspiPKS6 and AspiPKS7 from cyathea spinulosa, FhPKS2, FhPKS3, FhPKS4, FhPKS5 and FhPKS6 from ficus himalayana, and CrPKS from floating fern. The mutant is obtained by carrying out single point mutation on the second amino acid of AspiPKS7 or deleting an N-terminal redundant sequence. The polyketide synthase or the mutant is used for replacing NnHisspS in an original FBP luminescence system, and the biological self-luminescence level can be remarkably improved. The invention further provides a simplified and improved FBP system which comprises the polyketide synthase or the polyketide synthase mutant, CPH, H3H and Luz, and the application convenience and the application luminescence capacity of the FBP system are improved.

The invention discloses a glycosyl transferase UGT76G1 mutant and a method for catalytically synthesizing rebaudioside M by using the glycosyl transferase UGT76G1 mutant, and relates to the technical field of biological catalytic synthesis. The glycosyl transferase UGT76G1 mutant is any one of the following components (A)-(C): (A) a protein obtained by performing any one or more of the following mutations on the basis of an amino acid sequence as shown in SEQ ID NO.1: the 89th amino acid is mutated into H from M; the 380th amino acid is mutated from L to M; the 411th amino acid is mutated from A to Y; (B) a protein which has 95% or 98% or more of identity with the amino acid sequence defined by (A) and has the same function as the amino acid sequence defined by (A); and (C) a fusion protein obtained by connecting a tag to the terminal of the protein defined in (A) or (B). The glycosyl transferase UGT76G1 mutant disclosed by the invention is obtained by mutating and screening glycosyl transferase UGT76G1, has higher catalytic activity, and can be used for efficiently synthesizing rebaudioside M by taking rebaudioside D and UDPG as substrates.

The present invention belongs to the field of biological detection technology, specifically relating to a monoclonal antibody combination for detecting HPV18 E7 protein and its application. This combination consists of monoclonal antibodies 5F8 and 6G6, wherein the heavy and light chain variable regions of 5F8 and 6G6, respectively, contain specific complementarity-determining region sequences (as shown in SEQ ID NOs. 1 to 12), conferring high specificity for HPV18 E7 protein. Verification using a double-antibody sandwich ELISA and a biotin-avidin amplification system demonstrated that this antibody combination specifically recognizes HPV18 E7 protein, exhibits no cross-reactivity with other high-risk HPV oncoproteins, and exhibits a detection sensitivity of 1 ng / mL. This combination can be used to construct detection systems such as ELISA kits and immunochromatographic test strips, suitable for rapid screening of HPV18 E7 protein in cervical exfoliated cell samples.

The invention discloses a radish bolting and flowering gene RsSUF4 regulated by m6A demethylase and application of the radish bolting and flowering gene RsSUF4, relates to the technical field of bioengineering, and is technically characterized in that the gene is RsSUF4. The functions of the demethylase RsALKBH10B and the flowering inhibition gene RsSUF4 in the flowering process are analyzed in a targeted manner, and a specific association mechanism between m6A modification and radish flowering regulation and control is defined.

The invention relates to the field of gene engineering, and discloses a D-3-phosphoglycerate dehydrogenase mutant, a coding gene thereof and application of the D-3-phosphoglycerate dehydrogenase mutant and the coding gene. Compared with a protein with an amino acid sequence as shown in SEQ ID NO: 1, the mutant comprises mutation at the 131 site and / or the 285 site. The mutant can specifically catalyze a substrate 3-phosphoglycerate to synthesize 3-phosphate-hydroxypyruvic acid, can effectively promote synthesis of L-serine in a metabolic pathway of microorganisms, improves the yield and conversion rate of the L-serine, and reduces the yield of byproducts at the same time.

The invention provides a modified TnpB-omega RNA gene editing system and application of the modified TnpB-omega RNA gene editing system. In particular, the present invention provides a modified TnpB related [omega] RNA skeleton. The modified omega RNA skeleton provided by the invention has at least three stem-loop structure fragments, and has a shorter length and a compact structure than a natural omega RNA skeleton, so that the modified omega RNA skeleton and TnpB can be jointly carried in a single AAV vector, the administration efficiency is improved, and high-efficiency gene editing is realized.