22results about How to "Efficient expression" patented technology

The invention relates to a strong promoter P7 suitable for streptomyces and application thereof, and relates to the field of genetic engineering and microbial metabolism engineering. The nucleotide sequence of the strong promoter P7 is as shown in SEQ ID No.1, and the strong promoter comprises a plasmid vector of the strong promoter; a host cell comprising the plasmid vector; the invention also discloses application of the strong promoter, the plasmid vector and the host cell in starting expression of a target gene. Compared with the prior art, the characterization of the strong promoter P7 provides an effective tool element for streptomyces strong promoter engineering and high-efficiency gene expression, and has important significance on streptomyces silent gene characterization, high-efficiency gene expression, metabolite synthesis, metabolic pathway reconstruction and the like. The strong promoter can be applied to common streptomyces type strains, and has important significance on high yield of important proteins including enzymes and important metabolites from actinomycetes.

This invention discloses a stable transfected cell line for glucocorticoid detection based on the Gal4-DBD-GRLBD fusion protein of marine medaka and its construction method, belonging to the field of environmental toxicology detection technology. The construction method includes: designing specific primers to amplify the GR ligand-binding domain (LBD) of marine medaka, ligating it to the pBIND vector to form the coding sequence of the Gal4-DBD-GRLBD fusion protein, subcloning it into the Dualpromoter vector to construct the pBIND-omeGR expression plasmid; co-transfecting HeLa cells with the pBIND-omeGR plasmid, UAS reporter plasmid, and transposase plasmid at a mass ratio of 1.66:1:0.34, and obtaining the stable cell line HeLa-pBIND-omeGR expressing the fusion protein after selection with puromycin. The cell line exhibits ECMO activity against dexamethasone. 50 The value was 0.841 nM, and the induction activation factor was 2.34 times. This invention solves the technical problems of existing mammalian GR detection systems, which are difficult to accurately predict fish toxicity effects and have complex construction of full-length receptor systems. It has the advantages of simple construction, strong versatility, and good stability, and can be used for marine ecotoxicological assessment, environmental pollutant monitoring, and chemical safety evaluation.

The invention discloses a bacillus subtilis genome integrated plasmid for efficient and stable expression of genes and application of the bacillus subtilis genome integrated plasmid in production of beta-galactosidase. The plasmid comprises a bacillus subtilis promoter, an RBS sequence, a terminator, a bacillus subtilis resistance gene, a genome integration homologous arm, an escherichia coli replicon and a resistance gene. The homologous arm is selected from upstream and downstream sequences of sites behind termination codons such as genes spxA, tatCY, rlmCD and the like, so that exogenous genes are accurately integrated to a plurality of non-essential sites on the premise of not damaging essential genes. The invention also provides a method for constructing a bacillus subtilis engineering strain for producing beta-galactosidase by using the plasmid. The integration system is high in expression level, stable expression can be achieved without adding antibiotics and inducers, the fermentation cost is remarkably reduced, the enzyme production efficiency is improved, and the integration system is suitable for industrial enzyme preparation production.

The invention discloses an eurytopic chlamydomonas reinhardtii expression vector based on a PSAD promoter and a 3 * Flag tag as well as a construction method and application of the eurytopic chlamydomonas reinhardtii expression vector. The nucleotide sequence of the eurytopic chlamydomonas reinhardtii expression vector is shown as SEQ ID No.1; the vector comprises a PSAD promoter, a target gene insertion site EcoR V, a 3 * Flag tag at the 3'terminal, an rbcS2 terminator, a TUB2 promoter, a hygromycin B screening gene and an rbcS2 terminator. The construction method comprises the following steps: constructing a skeleton of a pHyg-PSAD-3Flag vector; constructing a sequence fragment fused with a 3 * Flag tag; and connecting the sequence fragment fused with the 3 * Flag tag with a plasmid skeleton fragment through T4 DNA (deoxyribonucleic acid) enzyme so as to construct the pHyg-PSAD-3Flag vector. According to the invention, the expression plasmid of the chlamydomonas pHyg-PSAD-3Flag can be used for efficiently screening algae strains capable of stably expressing target protein.

The application relates to the technical field of marketing, and discloses a marketing management method and system based on big data, which comprises a dynamic situation sensing and demand prediction module: potential consumption demand is predicted based on authorized multi-source data; an active situation declaration and solution generation module: structured life task declarations actively input by users are received, and a solution package is generated; an individual utility preview and verification module: a product is subjected to multi-physical field simulation and multi-dimensional adaptability evaluation; a closed-loop feedback and ethical guarantee module: the system is continuously optimized and data compliance is guaranteed. Through the cooperative operation of the modules, the demand identification mode can be optimized, the marketing precision can be improved in combination with a double path, invalid guidance can be reduced, the user interaction experience can be improved, the decision cost can be reduced, the cold start problem can be alleviated, data compliance can be guaranteed through a multiple privacy protection mechanism, user trust can be enhanced, long-term iterative optimization of the system can be realized, and operation stability can be guaranteed.

The application belongs to the technical field of genetic engineering, and particularly relates to a method for establishing a plant line of an influenza B virus HA protein expressed by Nicotiana benthamiana. According to the method, a nucleotide sequence coding the BV-HA protein is first connected to a plant binary expression vector pHREAC to construct a recombinant plasmid, then the recombinant plasmid is transformed into an E. coli competent cell, an Agrobacterium competent cell in sequence, and the Agrobacterium competent cell is used to infect the Nicotiana benthamiana, so that the BV-HA protein is obtained through transient expression in the Nicotiana benthamiana cell. The BV-HA protein purified through the method has high biological activity, and can be used for further biological function research. The method can realize one-step purification of the protein, is simple and fast, and has high protein purity. The exogenous recombinant protein obtained through the method can be used for biological safety evaluation and biological function research.

This invention provides a method and application for transient expression of collagen in plants. The collagen fragment C3S1, C3S2, C3S3, C3S4, and C3S5 are linked together with the amino acid sequences shown in SEQ1, SEQ2, SEQ3, SEQ4, and SEQ5, respectively. This collagen fragment can be expressed using tobacco as a bioreactor. By designing expression vectors, its expression effect in tobacco can be compared, and a transient expression system in tobacco can be established to achieve efficient expression of the collagen fragment in tobacco. This preparation method can express recombinant collagen fragments in tobacco with high expression levels. Compared to full-length intron-modified collagen or expression using type I collagen, this method achieves efficient expression of type III collagen fragments in tobacco bioreactors. It has a short cycle time, requires no maintenance of cell lines and bioreactors, significantly reduces production costs, and has a high safety index.

This invention belongs to the field of biomedical technology and discloses a chimeric antigen receptor targeting CD20, its gene expression construct, and its applications. The chimeric antigen receptor provided by this invention specifically includes an amino acid fragment with the sequence shown in SEQ ID NO:29 or a variant fragment having at least 85% sequence identity with SEQ ID NO:29. Its functional fragments, including the signal peptide, antigen-binding region, hinge region, transmembrane region, intracellular co-stimulatory domain, and signal transduction domain, form an organic whole, working synergistically to endow the chimeric antigen receptor with good CD20-targeting binding, T-cell activation, and cell-killing effects, demonstrating excellent application potential in the preparation of drugs for the treatment of relapsed / refractory B-cell malignancies.

This invention discloses an amidase gene for degrading amide alcohol antibiotics and amide herbicides. right here The amidase gene, along with its encoded protein and applications, has the nucleotide sequence SEQ ID NO.1, a full length of 912 bp, encoding 303 amino acids, and its amino acid sequence is SEQ ID NO.2. This invention relates to the gene... right here This is the first publicly disclosed amidase gene for amide alcohol antibiotics cloned from the genome of a single isolated strain. The amidase encoded by this gene can not only hydrolyze the amide bonds of amide alcohol antibiotics, but also degrade various amide chemical herbicides such as propargite, bromoxynil, chlorpyrifos, and chlorobromopropylate. The amidase encoded by this gene, as well as genetically engineered bacteria containing the amidase gene, have very important theoretical and applied value in the degradation of amide alcohol antibiotic pollutants in aquatic environments such as sewage treatment plants and livestock and poultry breeding wastewater, and in the remediation of herbicide-contaminated soil.

The application discloses a recombinant carrier for preparing HLA / beta 2M complex and a preparation method and application thereof, relates to the technical field of antibody preparation of MHC-polypeptide complex, and provides a recombinant carrier for preparing HLA / beta 2M complex, wherein FOS leucine zipper bZIP motif is inserted between NheI / AvrII enzyme cutting sites of an original carrier, JUN leucine zipper bZIP motif is inserted between BsiWI / BlpI enzyme cutting sites, beta 2M gene sequence is inserted between AgeI / NheI enzyme cutting sites, and HLA sequence is inserted between specific enzyme cutting sites, so that the prepared HLA / beta 2M complex has high expression, and the highest expression reaches 92.45 mg / L.

The invention belongs to the technical field of bioengineering, and particularly relates to an aspergillus niger chassis cell capable of efficiently expressing foreign protein, an expression system and a method for efficiently expressing foreign genes by using the aspergillus niger chassis cell. The Aspergillus niger chassis cell is constructed by integrating a PSST sequence containing a specific spacer sequence to a site beneficial to high expression of a foreign protein, so that the copy number of a foreign protein gene can be increased, the foreign protein can be prevented from being degraded, the integration efficiency of the foreign protein gene can be improved, the utilization rate of saccharides during fermentation can be increased, and the application prospect of the Aspergillus niger chassis cell is widened. Further, multi-copy exogenous genes can be efficiently integrated at one time, and the efficient expression of the exogenous genes is realized under the guidance of the strong promoter and the signal peptide.

The invention discloses a method for realizing transient expression of exogenous target genes in rhododendron molle plants by using a vacuum agrobacterium infection method, and belongs to the technical field of forest tree genetic engineering. The method comprises the following steps: performing ultrasonic pretreatment on rhododendron molle seedlings, immersing the plants into an agrobacterium infection solution containing a target gene, performing circulating vacuum treatment with specific parameters to enable the infection solution to efficiently permeate into tissues, and finally performing shading and normal culture to complete transient expression. By optimizing the ultrasonic treatment conditions and combining the circulating negative pressure vacuum infiltration technology, the problems that a traditional injection method is low in efficiency, tedious in operation and incapable of achieving batch treatment are solved. According to the method, the conversion efficiency and expression uniformity of the exogenous gene at the whole rhododendron molle plant level are remarkably improved, the operation is simple and convenient, the period is short, the repeatability is good, and an efficient and reliable technical platform is provided for gene function research and molecular breeding of rhododendron molle and rhododendron plants.

This invention discloses a recombinant Beauveria bassiana, its preparation method, and its applications. Firstly, this invention discloses a recombinant Beauveria bassiana capable of expressing parasitic bee venom proteins. It further discloses the preparation method of the recombinant Beauveria bassiana and its application in insecticidal applications. Based on the codon preference of Beauveria bassiana, this invention optimizes the gene sequence of parasitic bee venom proteins, expands the resource library of virulence factors for biocontrol, and integrates this sequence into Beauveria bassiana to obtain recombinant Beauveria bassiana, which can efficiently express parasitic bee venom proteins, overcoming the shortcomings of slow onset and weak toxicity of traditional biocontrol bacteria. Furthermore, the preparation method of the recombinant Beauveria bassiana is simple, and the obtained recombinant Beauveria bassiana is very stable, with continuous expression of exogenous virulence factors.

This invention relates to the field of animal immunology, and more particularly to an antigen that enhances the immune response to Newcastle disease virus (NDV), its preparation method, and its application. The antigen is a fusion protein obtained by expressing the Ii-key-F-Fc recombinant vector using engineered bacteria, named the Ii-key-F-Fc fusion protein. The Ii-key-F-Fc recombinant vector is obtained by splicing three major antigenic epitope fragments from the NDV F protein and then tandemly connecting them with chicken IgY Fc and Ii-key. This invention constructs the pCold-TF-Ii-key-F-Fc prokaryotic expression plasmid by tandemly connecting the novel chicken vector Ii-key-Fc with the F antigenic epitope, and then expresses the fusion protein using an E. coli expression system to obtain a new antigen that enhances the immune response in poultry. This provides a new approach for developing a novel NDV vaccine and a new strategy for improving the immune response of subunit vaccines in poultry.

The invention belongs to the technical field of enzyme engineering, and discloses a novel xanthine oxidase (XOD) gene cluster, a recombinant expression vector, a recombinant strain and application thereof. The gene cluster is a novel XOD gene cluster found in a Pseudomonas putida KT2440 genome for the first time through comparative analysis of bioinformatics, the gene composition mode of the gene cluster is remarkably different from that of a known XOD gene cluster, and a brand new gene resource and a technical path are provided for developing recombinant XOD with high performance and low cost. According to the present invention, with the recombinant expression vector constructed by replacing the recombinant expression vector with the strong promoter, the efficient expression of the novel xanthine oxidase can be achieved in the Pseudomonas putida KT2440;

The application discloses a recombinant herpesvirus of turkeys co-expressing classic strains and variant strains of IBDV VP2 and a construction method and application thereof, and belongs to the field of veterinary biological technology. The recombinant virus has good genetic stability, and has a consistent replication rate with HVT vaccine viruses in vitro. When used as a vaccine, 1-day-old chicks can be clinically protected against vvIBDV and nVarIBDV at 100% after single immunization, can induce the body to produce high-titer specific antibodies, protect the immune organs of chickens from virus invasion, and has a broad industrial application prospect in the preparation of IBDV multivalent vaccines.

The application provides a fusion protein for antibody site-specific coupling and application thereof. The fusion protein of the application is mainly fused by an antibody Fc affinity protein binding monomer and an avidin protein, realizes direct coupling of the antibody Fc affinity protein and a biotin-labeled molecule, and no longer uses a click chemistry scheme, so that the production and use difficulty of the antibody Fc affinity protein Protein G is greatly simplified, the Protein G protein no longer needs to be activated through the click chemistry in the production, the transportation and storage conditions no longer need to be strictly controlled, and even one-step antibody molecule coupling can be realized.

The application discloses an enhancer regulatory element for transcriptional regulation and expression of a human LTF gene and application thereof, wherein the sequence of the regulatory element is SEQ ID No. 1. Experiments prove that the human-specific regulatory element can significantly activate the expression of a reporter gene (up to 14.93 times) in a bovine mammary epithelial cell line (MACT), breaks through the limitation of a species-conserved element in the prior art, and an expression system designed based on the element can realize efficient expression of LTF at a human level, thereby providing a new tool for mammary gland bioreactors and development of functional dairy products.

This invention relates to the field of antimicrobial peptide technology, and particularly to the antimicrobial peptide Ple-AB, its preparation method, and its applications. Using the fungal defensin Plectasin as a template, this invention designs and modifies the antimicrobial peptide Ple-AB. Compared with Plectasin, this antimicrobial peptide exhibits significantly higher antimicrobial activity and better bactericidal effects against Gram-positive bacteria; it also shows higher stability, particularly excellent resistance to trypsin; and its fermentation expression level is significantly improved. Furthermore, this antimicrobial peptide has very low hemolytic activity and cytotoxicity, making it suitable for development in antimicrobial drugs, additives, cosmetics, health products, and other fields, with broad application value and market prospects.

This invention provides recombinant humanized elastin, its preparation method, and its applications, relating to the field of bioengineering technology. Based on humanized elastin, this invention utilizes a combination design based on the functional domains of elastin to obtain recombinant humanized elastin, discovering novel bioactive functional fragments of humanized elastin. The recombinant humanized elastin of this invention integrates multiple elastin functional sites. Specifically, this recombinant humanized elastin exhibits significant effects in promoting cell proliferation and increasing the expression of collagen and elastin genes, demonstrating high safety and wide applicability in various scenarios, and providing a new active ingredient for anti-aging related products. Furthermore, the theoretical molecular weight of this recombinant humanized elastin is significantly smaller than that of natural full-length elastin, reducing obstacles for elastin to cross the skin barrier and exert its biological functions, thus exhibiting excellent biological efficacy.

The present application relates to the field of bioengineering, and particularly relates to a mutant of mu-conotoxin and a preparation method and an enzyme composition thereof.The present application provides a mutant of mu-conotoxin, which has an amino acid sequence as shown in SEQ ID NO:1, or an amino acid sequence obtained by substituting, deleting or adding one or more amino acids in the amino acid sequence, and an amino acid sequence which is functionally identical or similar to the amino acid sequence, or an amino acid sequence which is at least 80% identical to the amino acid sequence.The technical route and process provided by the present application greatly improve the production efficiency and quality of mu-conotoxin, are suitable for industrial large-scale production, and have a wide application prospect.

The present application relates to ionizable lipid molecules containing a tocopherol structure, lipid nanoparticles comprising the same and uses thereof. Specifically, an ionizable lipid molecule containing a tocopherol and derivatives thereof structure represented by formula (1), lipid nanoparticles comprising the same, methods of preparation and uses thereof are provided. Compared with ionizable lipid molecules conventionally used in the art, the lipid nanoparticles prepared from the ionizable lipid molecules represented by formula (1) of the present application can significantly improve the delivery efficiency and expression of nucleic acids.