1573 results about "Recombinant expression" patented technology

The invention provides a thermal-stability glucose oxidase mutant, polynucleotide for encoding the glucose oxidase mutant, a recombinant expression vector containing the polynucleotide, a host cell for expressing the glucose oxidase mutant, and applications of the thermal-stability glucose oxidase mutant, the polynucleotide, the recombinant expression vector and the host cell in the fields of food, chemical industry, medicine, agriculture or feed.

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 the technical field of bioengineering, and discloses a mutant of Burkholderia lipase as well as a recombinant expression vector and application of the mutant, the mutant comprises P131S, T132A, T251A, T251S, P131A, P131Q, T132G, Q88S and P113A single processes; and a Q88S double process and a P113A double process; and a multi-site mutation combination: a combination I: Q88S, P113A, T132G and L287S, and a combination II: Q88S, P113A, T132G and L287S; a combination II: Q88S, P113A, P131S and L287S; and a combination III: Q88S, P113A, P131S and T251A. Based on experiments, the invention finds that each mutant can not only improve the protein yield, but also improve the enzyme activity by 57% at most; ePA enrichment experiments find that the EPA content of the mutant can be increased by 33% at most, and the mutant has excellent EPA purification and enrichment effects.

The invention belongs to the technical field of bioengineering, and particularly relates to recombinant III-type humanized collagen as well as a coding gene and application thereof. According to the invention, a novel III-type humanized collagen sequence is designed by adopting an advanced AI technology and molecular dynamics simulation, so that the recombinant III-type humanized collagen product is ensured to have high stability and degradation resistance. According to the invention, the screened new III-type humanized collagen sequence is transferred to the pichia pastoris strain for recombinant expression, and the prepared recombinant III-type humanized collagen has the advantages of strong stability, high expression quantity and good hydrophilicity, and meanwhile, the recombinant III-type humanized collagen also has obvious effects of promoting cell proliferation and cell adhesion.

The invention discloses a PgEGY3 gene for improving cold resistance of pennisetum alopecuroides and increasing fresh weight of leaves and application of the PgEGY3 gene, and relates to the technical field of genetic engineering, and the nucleotide sequence of the PgEGY3 gene of the pennisetum alopecuroides is shown as SEQ ID NO.1. The invention also discloses a protein coded by the Pennisetum alopecuroides PgEGY3 gene. The invention also discloses a recombinant expression vector and a recombinant host cell comprising the Pennisetum alopecuroides PgEGY3 gene. The invention also discloses application of the PgEGY3 gene, a recombinant expression vector and a recombinant host cell in improving cold resistance and biomass of pennisetum alopecuroides. It is verified that the Pennisetum alopecuroides PgEGY3 gene can improve the cold resistance of plants, and the growth state of the plants under cold stress is remarkably improved through overexpression of the PgEGY3 gene.

The invention relates to the field of gene engineering for enhancing the cold resistance of plants, and relates to a recombinant expression vector, a recombinant strain and a primer pair of a pennisetum purpureum PpMYB2 gene, a method for improving the cold resistance of the plants and application of related materials in improving the cold resistance of the plants. One purpose of the invention is to provide application of the Pennisetum purpureum PpMYB2 gene and related materials thereof in improving the cold resistance of the plant, the amino acid sequence of the encoding protein of the Pennisetum purpureum PpMYB2 gene is as shown in SEQ ID NO.1, and the plant is arabidopsis thaliana or Pennisetum purpureum. Under the background that extreme low temperature events caused by global climate change are increasingly frequent, the cold resistance enhancing technology provided by the invention has important significance on guaranteeing the stability of agricultural production and reducing loss caused by low temperature. Through overexpression of the gene PpMYB2, the adaptability of perennial pennisetum purpureum to low-temperature stress can be remarkably improved, so that the planting range of perennial pennisetum purpureum is expanded to regions with lower temperature, and the survival and growth performance in different ecological environments is enhanced.

The invention provides a recombinant vector for expressing LPL, a recombinant bacterium and an LPL production method, and belongs to the technical field of gene engineering. The invention provides a recombinant expression vector of Pseudomonas aeruginosa lipoprotein lipase LPL, wherein an LPL coding gene is derived from Pseudomonas aeruginosa. The recombinant expression vector is introduced into a pseudomonas aeruginosa PAO1 cell, an expression strain is successfully constructed, the expression quantity of target protein is improved, efficient expression of LPL in PAO1 is achieved, and the produced LPL is high in enzyme activity and wider in application range.

The invention discloses a camel source single-domain antibody GH12 for recognizing a conformation B cell epitope in clostridium difficile glutamate dehydrogenase and application of the camel source single-domain antibody GH12. The nucleotide sequence of the camel source single-domain antibody GH12 is shown as SEQ ID NO: 1, and the amino acid sequence of the camel source single-domain antibody GH12 is shown as SEQ ID NO: 2. The camel source single-domain antibody GH12 disclosed by the invention has the advantages of strong specificity, high sensitivity, good accuracy and the like, and it is proved that the camel source single-domain antibody GH12 can be used for detecting recombinant expressed CD-GDH and endogenous CD-GDH by enzyme-linked immunosorbent assay (ELISA), protein dot blot hybridization (DB) and other methods.

The invention belongs to the technical field of bioengineering, and particularly relates to recombinant III-type humanized collagen and application thereof. Area fragments with good hydrophilicity in a human III-type collagen alpha chain are selected and sequentially connected to serve as expression protein, the amino acid sequence is 100% identical to that of a corresponding part of natural human III-type collagen, and immunological rejection cannot be generated when the human III-type collagen is applied to a human body. According to the invention, the screened new III-type humanized collagen sequence is transferred into pichia pastoris for recombinant expression, the prepared recombinant III-type humanized collagen has the advantages of strong stability, high expression quantity and good hydrophilicity, and meanwhile, the recombinant III-type humanized collagen also has obvious effects of promoting cell proliferation and cell adhesion.

The invention discloses humanized III-type collagen as well as a preparation method and application thereof, and belongs to the technical field of bioengineering. According to the method, firstly, on the basis of an original humanized III-type collagen gene, cell adhesion activity is taken as an evaluation index, a better cell adhesion sequence is screened out and spliced and integrated with the original collagen gene, and brand-new humanized III-type collagen is obtained. And constructing a recombinant expression vector of the humanized III type collagen containing the adhesion sequence by taking the probiotic EcN as a host cell, and introducing the recombinant expression vector into the EcN to obtain a strain of functional recombinant probiotic. Compared with a traditional escherichia coli engineering bacterium BL21 (DE3), the recombinant probiotics have good cell adhesion activity while efficiently expressing the humanized type III collagen, the bacterial endotoxin content of the recombinant humanized type III collagen obtained through purification meets the industrial standard, and the recombinant probiotics have good application prospects. The purification cost caused by industrial removal of endotoxin can be avoided, so that the production cost is reduced, and the method has a good application prospect.

Provided are an anti-BCMA single-domain antibody, and a preparation method therefor and the use thereof. Specifically, provided is a single-domain antibody having an amino acid sequence of SEQ ID No. 1. The single-domain antibody has high affinity, can thoroughly specifically target BCMA-positive cells, and can be applied to the detection of BCMA expression in bone marrow cells of MM patients. The single-domain antibody can be prepared into a specific antibody drug clinically used for preventing and treating BCMA-target-related diseases (such as multiple myeloma, B-cell acute lymphoblastic leukemia, non-Hodgkin's lymphoma and Hodgkin's lymphoma); or a BCMA protein detection kit, etc. The single-domain antibody has a stable structure, a small molecular size, is easily recombinantly expressed and has a low production cost, can be used alone or as a drug delivery system to carry relevant drugs, and has very wide prospects and important significance in fields such as drug application and clinical diagnosis.

The invention discloses a collagenase mutant, a gene segment, a recombinant plasmid, a recombinant expression system, a collagen peptide, and a preparation method and application thereof, and belongs to the field of gene engineering and fermentation engineering. The collagenase mutant KT513 with high specific activity is obtained through recombinant expression in a trichoderma reesei host, and the enzyme activity of the collagenase mutant KT513 reaches 7423.18 U / mL. The optimum action pH of the enzyme is 3.0, the red snapper scale collagen can be subjected to enzymolysis in the pH range of 3.0-5.0, and the enzyme has excellent enzymolysis activity. After the collagenase mutant KT513 is subjected to enzymolysis at 40 DEG C for 4 hours, the extraction rate of fish scale collagen reaches 80.09%, and the prepared collagen peptide has good antioxidant activity. When the addition concentration is 10 mg / mL, the in-vitro scavenging activity of the prepared collagen peptide on hydroxyl radicals reaches 99.26% and is equivalent to that of ascorbic acid, and the collagen peptide has a wide application prospect in preparation of antioxidant preparations.

The invention discloses SGNH family esterase SH2 screened in a Tibetan hot spring environment and application of the SGNH family esterase SH2, and belongs to the technical field of gene engineering. According to the invention, a new SGNH family esterase gene is screened from a Tibetan hot spring environment, and recombinant expression is realized in escherichia coli. The recombinant esterase prepared by the invention has the characteristics of thermal stability, salt resistance, organic solvent resistance and the like, can be safely applied to tooth whitening or skin exfoliating products, avoids damage of traditional chemical bleaching agents to enamel, and has important application value in the fields of daily chemicals, medicines, food processing and the like.

The invention provides a BaP4H mutant and application thereof, and belongs to the technical field of enzyme engineering, the BaP4H mutant is mutated on an amino acid sequence as shown in SEQ ID NO.1; the invention further provides a recombinant expression vector for expressing the BaP4H mutant, a recombinant cell, a preparation method and application. According to the BaP4H mutant provided by the invention, the recombinant collagen proline is efficiently hydroxylated, and the stability of the recombinant collagen is enhanced.

The invention relates to the technical field of gene engineering, in particular to a cat serum albumin preparation and characterization method. The cat serum albumin gene sequence is optimized, a recombinant expression vector is successfully constructed, pichia pastoris genetically engineered bacteria are prepared, high-yield expression of the cat serum albumin gene in pichia pastoris is achieved, a purification system is established and perfected, and a pure product with the purity larger than 98% is obtained through affinity exchange column purification. Compared with the natural cat serum albumin, the cat serum albumin has a secondary structure, stability and antioxidant activity which are consistent with those of the natural cat serum albumin, has the characteristic of low hemolytic activity, can realize large-scale efficient production, can be applied to development of pet foods and medicines, and has wide application value and market prospect.

The present invention relates to a recombinant human type IV collagen and a preparation method therefor, which belong to the technical field of bioengineering. A recombinant human type IV collagen is designed on the basis of a partial sequence in an α2 chain of human type-IV collagen, and DNA encoding of the protein is ligated into an expression vector to construct a recombinant expression vector; then the recombinant expression vector is transformed into competent Escherichia coli cells, and a recombinant expression plasmid is extracted; after the plasmid is linearized, the linearized plasmid is transformed into an engineered strain HCPB-PPKEX2 to obtain a recombinant engineered strain; and the engineered strain is cultured, the expression of a target protein is induced, and the target protein is purified and identified. It is further demonstrated experimentally that the collagen can be completely cleaved by Kex2, Ste13 and CPB enzymes and efficiently secreted and expressed extracellularly, and the obtained sequence is completely consistent with a theoretical sequence. It is also verified experimentally that the recombinant collagen has a cell adhesion activity and a cell migration-promoting activity. The method avoids the risk of residual exogenous proteins, shortens the time and reduces the cost of the subsequent purification process, and thus has a relatively high safety and application value.

The invention discloses application of scylla serrata antibacterial peptide Scycin in preparation of an anti-inflammatory composition and a preparation method of the scylla serrata antibacterial peptide Scycin. The nucleotide sequence of the scylla serrata antibacterial peptide Scycin is shown as SEQ ID NO.01. The invention further discloses a preparation method of the scylla serrata antibacterial peptide Scycin. In an LPS (lipopolysaccharide)-induced RAW 264.7 macrophage inflammation model, the recombinant expression plasmid vector containing the scylla antibacterial peptide Scin remarkably inhibits an inflammation cascade reaction by down-regulating expression of proinflammatory factors such as TNF-alpha (tumor necrosis factor-alpha) and IL-6 (interleukin-6), the anti-inflammatory effect of the recombinant expression plasmid vector is equivalent to that of a mesalazine positive control group, and obvious cytotoxicity is not observed.

The invention relates to the technical field of biomedicine, discloses mutant photo-induced ion channel ChR-2 protein, and further discloses a nucleic acid construct, an expression vector, a cell, related application and a computer model. The light-sensitive channel ChR2 protein mutant obtained by the invention has stronger light current, and the light-sensitive capability of the light-sensitive channel ChR2 protein mutant is at least improved by 100 times; besides, the invention also obtains a nucleotide sequence for coding the light-sensitive channel ChR2 protein mutant, constructs a recombinant expression vector, and obtains the light-sensitive channel ChR2 protein with higher expression quantity and stronger light sensitivity, and the light-sensitive channel ChR2 protein is very suitable for expression in cells of mammals (especially human); according to the invention, the rhodopsin in different channels is systematically studied by modifying the position G224 in the helix 6 of seven transmembrane helix motifs, which proves that the mutation of the position G224 in the helix 6 in WT ChR2 accelerates the photosensitivity of the channels, and the considered position is homologous in the helix 6 of the rhodopsin in different channels.

The current disclosure provides binding polypeptides (e.g., antibodies), and effector moiety conjugates thereof (e.g., antibody-drug conjugates or ADCs), comprising a site-specifically engineered drug-glycan linkage within native or engineered glycans of the binding polypetpide. The current disclosure also provides nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided.

The invention belongs to the technical field of agricultural microorganisms, and particularly relates to a method for identifying a compound for inhibiting synthesis of vomitoxin from wheat stem rot pathogen. The method comprises the following steps: cloning an open expression cassette of a key catalytic enzyme gene FpgTri4 in a process of biologically synthesizing vomitoxin by pathogenic bacteria, constructing recombinant expression plasmids from the open expression cassette, and transduction the recombinant expression plasmids into a Fusarium pseudograminearum strain to induce expression to generate toxosome; and the inhibitor with inhibition on the generation of vomitoxin can inhibit the generation of toxosome in the recombinant strain, so that the related inhibitor can be screened by using the method. The method disclosed by the invention is rapid, simple and easy to implement, can be used for high-throughput screening, explores compounds capable of preventing diseases and controlling toxicity, and has important significance on scientific prevention and treatment of wheat stem rot.

The invention relates to a lipase mutant and application thereof in preparation of epoxide, in particular to a lipase mutant which is derived from Tilletia indica and has remarkably improved peroxidation activity, a coding nucleic acid of the lipase mutant, a recombinant expression vector and a recombinant expression transformant containing a nucleic acid sequence, and an application of the lipase mutant in preparation of epoxide. The recombinant lipase or the recombinant expression transformant is used as a catalyst for preparing peroxy acid, and the peroxy acid reacts with olefin to generate epoxide. Compared with other methods (including a chemical method or a biological method and the like) for preparing alpha-epoxy pinane, the enzyme and the technical method thereof disclosed by the invention have the advantages of mild reaction conditions, environmental friendliness, lower cost, simplicity and convenience in operation, easiness in amplification and the like, and have a very good application prospect in the field of producing epoxy products by taking carbon-carbon double bonds as substrates.

The invention discloses a novel biosynthesis path for producing 3 '-phosphoadenosine-5'-phosphosulfuric acid and application, and belongs to the technical field of bioengineering. According to the invention, through recombinant expression and purification of ribose kinase, ribose phosphate pyrophosphate kinase, ribose phosphate transferase, adenine thiotransferase, adenosine phosphate sulfate kinase and polyphosphate kinase 2, a six-enzyme cascade reaction taking ribose and adenosine as substrates in vitro is constructed, the molar conversion rate reaches 22.85%, and the escherichia coli and bacillus subtilis PAPS cell factory can be used for preparing the recombinant ribose phosphate pyrophosphate kinase, the ribose phosphate transferase, the adenine thiotransferase, the adenosine phosphate sulfate kinase and the polyphosphate kinase 2. The yield of PAPS in a 5-L fermentation tank reaches 7.60 g / L and 5.03 g / L respectively. The PAPS synthesis route provided by the invention provides an extensible, economical and efficient solution for the production of sulfated compounds.

The invention discloses a double-structural-domain recombinant human fibronectin, the sequence of which is shown as SEQ ID NO.3, and the double-structural-domain recombinant human fibronectin is formed by fusing a collagen binding structural domain and a cell binding structural domain of human fibronectin. The invention also provides a recombinant expression vector containing the protein coding gene and a host cell. The protein is efficiently expressed through eukaryotic cells and engineering bacteria escherichia coli, and a fermentation and purification process suitable for an industrial scale is established in a matched manner. Functional experiments prove that compared with commercially available fibronectin products with a single structural domain, the double-structural domain recombinant human chemical fibronectin disclosed by the invention can remarkably promote proliferation, migration, adhesion and differentiation activity of human skin fibroblasts, and has wide application potential in the fields of skin repair, skincare and biomedical materials.

The invention discloses coprinus cinereus sourced laccase as well as a recombinant expression strain and a high-efficiency expression method thereof, and belongs to the technical field of microbiology and bioengineering. According to the invention, aspergillus niger MA70.15 is used as an experimental strain, and a random integration strategy is adopted, so that a pC3-PcitA-SPGlaA-lcc9-TtrpC-pyrG expression vector is integrated in an upstream region of a promoter of host bacteria citrate synthase in a form of nine continuous copies. In order to explore the influence of particles and osmotic regulators in a fermentation culture medium on laccase production through fermentation of recombinant aspergillus niger, the particles and osmotic regulators of different types and concentrations are added, so that the enzyme activity of the laccase fermented in a shake flask of the recombinant strain AnRcgL1 reaches 1566.7 U / L, the enzyme activity fermented in a 1L fermentation tank reaches 1961 U / L, and the recombinant high-efficiency expression of the laccase Lcc9 in the aspergillus niger is successfully realized.

The invention discloses a method for promoting lactobacillus activity enhancement, which comprises the following steps: S1, amplifying a target gene corresponding to an LPxTG structural protein, and amplifying a target gene corresponding to a signal peptide; s2, obtaining a target gene connected with the LPxTG structure and the signal peptide; s3, connecting the target gene obtained in the step S2 with a plasmid gene sequence to construct a recombinant expression vector; s4, introducing the recombinant expression vector obtained in the step S3 into escherichia coli BL-21 so as to increase the number of the recombinant expression vector; s5, introducing the recombinant expression vector obtained in the step S4 into lactobacillus plantarum C8 to obtain a recombinant strain; and step S6, co-culturing lactobacillus plantarum C8 and the recombinant strain obtained in the step S5. The lactobacillus plantarum C8 has the advantages that the whole lactobacillus plantarum C8 and the recombinant strain have better survival condition and adhesiveness in gastric juice and intestinal juice.

The invention belongs to the technical field of gene engineering, and particularly relates to a potato StCBF3 gene, a recombinant expression vector and application and a method thereof. The nucleotide sequence of the potato StCBF3 gene provided by the invention is as shown in SEQ ID NO.1, and the amino acid sequence of the protein expressed by the potato StCBF3 gene is as shown in SEQ ID NO.2. By constructing a potato transgenic line of the StCBF3 gene of an overexpression and silent expression CBF / DREB1 subfamily, physical and chemical indexes of the transgenic line and a wild type line under abiotic stress conditions such as high temperature, low temperature, high salt and drought are analyzed, and the effect of the StCBF3 gene in resisting abiotic stress of potatoes is disclosed; the problem that in the prior art, the resistance of potatoes to various different abiotic stresses cannot be improved at the same time is solved.

The present invention discloses an antibacterial peptide cecropin A mutant, a recombinant expression vector, a Bacillus subtilis engineering bacterium and applications thereof. By modifying the antibacterial peptide cecropin A, the present invention screened mutants with antibacterial activities superior to those of the original cecropin A, and through the construction of a recombinant expression vector, heterologous expression was carried out using the biosafe strain Bacillus subtilis. Compared with the original antibacterial peptide cecropin A, the antibacterial activities of the mutants of the present invention were increased by 60% and 80% respectively. The present invention uses Bacillus subtilis as a secretion expression system to express the antibacterial peptide cecropin A mutant, which is conducive to large-scale industrial production and cost savings.

Disclosed herein are modified strains for improving secretion of recombinantly expressed products secreted from a host organism with improved growth and productivity characteristics, as well as methods of using the modified strains.

The present invention relates to the field of genetic engineering technology, specifically disclosing a raccoon dog interferon-α and β-defensin fusion protein, its encoding gene, and its application. The raccoon dog interferon-α and β-defensin fusion protein has the amino acid sequence shown in SEQ ID NO:1. The present invention also provides a gene encoding the fusion protein, a recombinant expression vector, and a host cell. The fusion protein provided by the present invention possesses the dual biological activities of raccoon dog interferon-α and β-defensin, and has certain antiviral and antibacterial activities.

The present disclosure concerns polypeptide having fumonisin esterase activity exhibiting increased fumonisin esterase activity, when measured at a temperature of at least 37° C., when compared to the control polypeptide comprising the amino acid of SEQ ID NO: 3. The polypeptides of the present disclosure include one or more amino acid variations which contribute to the increase fumonisin esterase activity. The present disclosure also includes compositions comprising the polypeptide having fumonisin esterase activity, methods for detoxifying a fumonisin mycotoxin as well as processes for making the compositions comprising the polypeptide having fumonisin esterase activity.