96results about "Polypeptide with GST-tag" patented technology

Provided is a method for biosynthesis of a human body structural material type-VIII collagen. Also provided is a collagen, comprising one or more repeating units, wherein the repeating units are connected directly or by means of linkers, and each repeating unit comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 4, 7, 10, 13, 16, 19, 22, 25 or 28, or a variant thereof. The collagen can be used for biological dressings, human body bionic materials or plastic surgery materials. According to the method, the type-VIII collagen is produced by utilizing a genetic engineering technology, thereby overcoming the defects in the prior art.

The invention provides a preparation method, application and the like of large intestine expression recombinant A-type botulinum toxin (BONT / A). The recombinant BONT / A nucleic acid expression cassette comprises a tag, a restriction enzyme cutting site and a BONT / A nucleic acid sequence, and an exogenous protease recognition site is not introduced between a BONT / A light chain and a BONT / A heavy chain. The invention also provides a recombinant vector containing the nucleic acid expression cassette, a recombinant bacterium, a coded and expressed protein and the like, the prepared recombinant BONT / A does not introduce exogenous amino acid or only introduces two amino acids at one position, and the consistency of the recombinant BONT / A with the natural A-type botulinum toxin is ensured to the greatest extent. Through novel molecular design and preparation process, the recombinant reBONT / A with higher toxicity and low immunogenicity is efficiently obtained, and wide industrial application is facilitated.

The invention is applicable to the technical field of molecular biology, and provides a polypeptide specifically bound with a transferrin receptor, the polypeptide comprises a general formula of H1-H2-E1-E2-H3-H4-E3, and H1, H2, H3 and H4 independently comprise an alpha helical domain; the E1, the E2 and the E3 respectively and independently comprise a beta folding structural domain; when the polypeptide is combined with the hTfR1, transferrin is allowed to be combined to the hTfR1; the polypeptides are capable of binding and delivering oligonucleotides. The invention also provides a recombinant nucleic acid encoding the polypeptide, an expression vector comprising the recombinant nucleic acid, a recombinant host cell comprising the polypeptide, the nucleic acid and / or the expression vector, a conjugate of the polypeptide and oligonucleotide, and a corresponding pharmaceutical composition. The polypeptide disclosed by the invention can be highly specifically combined with hTfR1, so that accurate delivery of oligonucleotide is realized, and a practical basis is provided for treating diseases related to gene mutation.

The invention relates to Escherichia coli for producing 2 '-fucosyllactose as well as a construction method and application of the Escherichia coli, and belongs to the technical field of genetic engineering. A CRISPR / Cas9 system is used for transforming a genome of escherichia coli MG1655, a wcaJ gene and a lacZ gene are knocked out, the utilization rate of lactose and GDP-L-fucose in a metabolic pathway is increased, a promoter of a lacY gene is replaced with PJ23119, an alpha-1, 2-fucosyltransferase mutant with a dissolution promoting tag GST added to the N end is heterologously introduced, the expression quantity of the alpha-1, 2-fucosyltransferase mutant is increased through multi-copy integration, and the expression quantity of the alpha-1, 2-fucosyltransferase mutant is increased. Meanwhile, a gmd gene, an fcl gene, a manB gene and a manC gene are co-expressed at proper sites, so that the synthesis route of the GDP-L-fucose is optimized. The strain obtained through transformation can efficiently convert lactose into 2 '-FL and has the advantages of being low in metabolic burden, sufficient in precursor supply, high in enzyme stability and the like, the construction method is clear in step and suitable for industrial production, the yield of 2'-FL can be remarkably increased through microbial agents and fermentation application, and the production cost is reduced.

Provided are a single-chain polypeptide which is highly activated after entering a specific use site, attenuated and can be safer and more convenient to produce, and the corresponding use thereof, a nucleic acid encoding same and a preparation method therefor. The single-chain polypeptide comprises: a first domain, an intermediate amino acid sequence region, and a second domain, wherein the intermediate amino acid sequence region is located between the first domain and the second domain, and contains at least one first enzyme cleavage site, the first enzyme cleavage site is designed to be capable of being highly specifically cleaved by a first protease present in a specific part of the human body, and the single-chain polypeptide is converted into an activated form capable of inhibiting the release of an information substance after being cleaved by the first protease at the first enzyme cleavage site.

Provided are a Blumea balsamifera monoterpene synthase BbTPS3 and related biological materials thereof and use thereof. BbTPS3 is: A1) a protein having the amino acid sequence shown in SEQ ID NO: 2; A2) a fusion protein obtained by linking protein-tags at the N-terminus or / and the C-terminus of the protein shown in SEQ ID NO: 2; and A3) a protein having at least 90% identity and the same function as the protein shown in A1), which is obtained by performing substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence shown in SEQ ID NO: 2. BbTPS3 can catalyze GPP to form l-borneol, and can be used to regulate and produce plant monoterpene compounds and cultivate Blumea balsamifera (L.) DC.

The invention relates to a method for identifying and quantifying a polypeptide from a library of polypeptides. The method comprises the steps of: 1 - providing a polypeptide library and a detection tag library, 2 - generating a nested library comprising the polypeptides and the detection tags, 3 - sequencing the nested library, 4 - selecting a member of the nested library in one or several selection steps that are independent of a physical genotype-phenotype linkage, 5 - isolating the detection tag from the selected polypeptide, 6 - identifying and quantifying the detection tag by mass spectrometry, 7 - obtaining the sequence of the selected polypeptide.

The invention provides a human FGFR3 (Fibroblast Growth Factor Receptor 3) antigen expression recombinant yeast vaccine as well as a construction method and application thereof. A specific primer is designed to amplify an FGFR3 gene, a pEGKT316-FGFR3 recombinant plasmid is constructed, a saccharomyces cerevisiae YEF473A strain is transformed, and the vaccine is prepared through induced expression and thermal inactivation. The vaccine can effectively activate DC cell maturation (CD80 + cell proportion reaches 78.7%) and promote Th1 type cell factor secretion (IL-12p70 reaches 5889pg / mL), and shows a remarkable tumor inhibition effect (plt; 0.05), and a novel specific treatment means is provided for postoperative immunotherapy of bladder cancer.

Provided is a low-molecular-weight collagen having a 164.88° triple-helix structure. The present invention relates to small-molecule collagen, a preparation method therefor, and a use thereof. The collagen has an amino acid sequence represented by SEQ ID NO: 1 or a variant amino acid sequence after mutation of the amino acid sequence. The variant amino acid sequence retains the function of the amino acid sequence represented by SEQ ID NO: 1. The collagen has a short amino acid sequence and good transdermal absorption performance.

The present application provides multivalent protein scaffolds useful as therapeutic agents and useful for identifying novel therapeutic compounds. The invention also relates to multi-domain polypeptide constructs having a plurality of binding domains and a domain. The invention also provides a method for identifying a new candidate therapeutic agent by using the multivalent protein scaffold provided by the invention and the identified new therapeutic agent.

The present invention provides a recombinant botulinum toxin and a preparation method therefor. A nucleic acid encoding the recombinant botulinum toxin comprises a first nucleic acid fragment, a second nucleic acid fragment, a first modification tag and a second modification tag, wherein the first nucleic acid fragment encodes a heavy chain of botulinum toxin, the second nucleic acid fragment encodes a light chain of botulinum toxin, the first modification tag encodes a His tag, and the second modification tag encodes a GST tag or an MBP tag.

The invention relates to a method for preparing a nano metal iron oxide material from iron ion binding protein with specific binding capacity, and belongs to the technical field of genetic engineering. In order to overcome the technical problems of non-specific binding of metallothionein and iron ions and relatively low production efficiency in a traditional nano # imgabs0 # biological preparation method, the invention provides an iron ion binding protein with specific binding capacity, and a specific binding iron ion polypeptide sequence is connected on the basis of metallothionein, so that the iron ion binding protein with specific binding capacity is obtained. The specific binding amount of iron ions is increased, and the specific binding iron ion protein is used as a template to prepare the # imgabs 1 # nano material. According to the method for preparing the nano-metal iron oxide material through specific binding of the iron ion protein, reaction conditions are mild, # imgabs2 # is prepared and synthesized in an oriented mode through high selectivity of the specific binding iron ion protein, the preparation process is easy to control, and the particle size range of the prepared # imgabs3 # is 20-1000 nm.

The invention relates to application of a small peptide SsSCP in improvement of plant disease resistance. The small peptide SsSCP is prepared by the following steps: connecting a gene of the small peptide SsSCP to an expression vector to prepare an SsSCP recombinant expression plasmid; transferring the SsSCP recombinant expression plasmid into an expression host to obtain an expression strain; performing induced culture on the expression strain to obtain an induced expression culture; and purifying the small peptide SsSCP from the induced expression culture. The expression and purification method of the small peptide SsSCP is optimized, the small peptide GST-SsSCP is synthesized and purified through microorganisms, and meanwhile the activity of the SsSCP is kept. By applying the GST-SsSCP protein expressed and purified in vitro, ROS outbreak of different plants can be triggered, and the GST-SsSCP protein has broad-spectrum MAMP activity. And the disease resistance of plants is improved. As a green and environment-friendly biological prevention and control means, the protein has a remarkable effect on enhancing the resistance of tomatoes and soybeans to sclerotinia sclerotiorum and the resistance of tomatoes to bacterial leaf spot.

The present invention provides a method for producing a block copolymer, the method including a step of mechanochemically treating a mixture containing a protein and a molecule capable of plasticizing a protein.

The invention discloses a protein post-translational modification detector based on a GST fusion histone H3 fragment mixture as well as a preparation method and application of the protein post-translational modification detector. The protein post-translational modification detector comprises 9 segmented fragments H3F1-H3F9 of histone H3. The prepared GST is fused with an H3 fragment (H3F1-H3F9) mixed plasmid; the mixed plasmid is transformed into BL21-DE escherichia coli; GST fusion H3 fragment mixture expression is carried out; purification is carried out; after the mixture is combined with GSH resin, incubation with a cell lysis solution is carried out for PTM; the GST label of the mixture after PTM is cut off by using precession TM protease, and the H3F1-H3F9 mixture is released into a solution; if a signal peak of the corresponding molecular weight is detected in the mass spectrum, PTM of the organic acid can be confirmed.

Provided is a biosynthesis-based preparation method for a structural material of the human body. A polypeptide of the present application contains the structure of a C-terminal region of (repeating unit)n or (repeating unit)n-; the repeating unit contains an amino acid sequence as shown in SEQ ID NO. 1. Recombinant type V humanized collagen prepared in the present application has high activity in promoting cell adhesion, and does not cause an immune response when applied to the human body; moreover, the preparation method is novel, allows for large-scale production of the recombinant type V humanized collagen, and is widely used in the preparation of structural materials of the human body. The application field includes preparation of high-end medical devices, such as biological dressings, human body biomimetic materials, plastic-beauty materials, organoid culture, cardiovascular stents, coatings, tissue injection filling, ophthalmic materials, gynecological biomaterials, nerve repair and regeneration, liver tissue and vessel repair and regeneration, and 3D printing of artificial organ biomaterials.

The invention relates to a deubiquitination enzyme Fc fusion protein, a product thereof, a preparation method and application thereof in Miao compound screening, and belongs to the technical field of fusion proteins. The technical problem to be solved is that OTUD3 protein is unstable and easy to degrade in the prior art; according to the key point of the technical scheme, the OTUD3 and Fc fusion protein is provided, and the fusion protein is easy to purify while maintaining the activity of OTUD3 and can be applied to screening of Miao compounds.

The invention provides a method for prokaryotically expressing cat allergen Fel d 1 recombinant protein, which comprises the following steps of: cloning a cat allergen Fel d 1 gene to multiple cloning sites of an expression vector pGEX-4T-1 to obtain a recombinant expression vector pGEX-4T-1-Fel d 1, transforming the recombinant expression vector into escherichia coli Rosetta (DE3) competent cells to construct recombinant engineering bacteria pGEX-4T-1-Fel d 1 / Rosetta (DE3), and carrying out prokaryotic expression on the recombinant engineering bacteria pGEX-4T-1-Fel d 1 / Rosetta (DE3) to obtain the cat allergen Fel d 1 recombinant protein. IPTG (isopropyl-beta-d-thiogalactoside) is used for inducing protein expression, and a GST affinity chromatography medium is used for capturing high-purity soluble GST-Fel d 1 fusion protein. According to the invention, a pGEX-4T-1-GST label system is combined with Escherichia coli Rosetta (DE3), so that the problem of soluble expression of the cat allergen Fel d 1 protein is fundamentally solved.

The invention discloses a cell strain capable of stably expressing a multi-protein tag as well as a construction method and application of the cell strain, and belongs to the technical field of biology. The cell strain is transfected by a lentiviral vector, a fusion expression vector containing P2A self-cleavage polypeptide and a plurality of protein tag coding sequences is integrated into a host cell, and a stable expression cell strain and a fusion protein expressing a plurality of protein tags including green fluorescent protein are obtained through antibiotic screening. The cell strain can be used for antibody specificity verification, protein interaction research and multi-label system standardization experiment platforms, and has the advantages of stable expression, simple operation, reliable result and the like.

Provided is an immunogenic Mycoplasma pneumoniae polypeptide epitope, which is a polypeptide epitope derived from the key adhesion proteins P1, P30, P40 / 90 and P116 of type I and type II Mycoplasma pneumoniae, and / or the CARDS toxin thereof. Further provided is a recombinant protein, which contains one or more of the immunogenic Mycoplasma pneumoniae polypeptide epitopes. Further provided is a vaccine for preventing Mycoplasma pneumonia infections, which vaccine provides protection against at least one of type I and / or type II Mycoplasma pneumonia infections, and is characterized in that the vaccine contains the recombinant protein and is a vaccine capable of providing safe and effective protection against Mycoplasma pneumonia infections.

To provide agents and their use in the treatment of endocrine, nutritional and / or metabolic diseases in a mammal.SOLUTION: The present disclosure provides novel peptides which stimulate β-cell proliferation, have ability to rescue β-cells from apoptosis induced by glucotoxic conditions, and stimulate insulin secretion from rat INS-1 β-cells as well as isolated mouse pancreatic islets.SELECTED DRAWING: Figure 1

Provided is a method for producing a block copolymer, the method including a step for mechanochemically treating a mixture containing a protein and a molecule capable of plasticizing the protein.

The present invention provides a recombinant botulinum toxin and a preparation method therefor. A nucleic acid encoding the recombinant botulinum toxin comprises a first nucleic acid fragment, a second nucleic acid fragment, a first modification tag and a second modification tag, wherein the first nucleic acid fragment encodes a heavy chain of botulinum toxin, the second nucleic acid fragment encodes a light chain of botulinum toxin, the first modification tag encodes a His tag, and the second modification tag encodes a GST tag or an MBP tag.

The present disclosure provides, in part, compositions and methods for transiently removing neutralizing antibodies to AAV vectors, expanding the patient population suitable for gene therapy and, in addition, AAV re-medication / re-administration in patients previously treated with AAV vectors.

Presented herein are synthetic protein phosphorylation signaling networks.

The invention discloses a preparation method and application of a pseudosciaena crocea antibacterial peptide Elabela, a pseudosciaena crocea antibacterial peptide gene Elabela is obtained through gene cloning, and the nucleotide sequence of the pseudosciaena crocea antibacterial peptide gene Elabela is shown as SEQ ID NO.1. The antibacterial peptide Elabela is further prepared through prokaryotic expression and purification, and the amino acid sequence of the antibacterial peptide Elabela is as shown in SEQ ID NO. 2. The antibacterial peptide Elabela prepared by the invention has broad-spectrum antibacterial activity, and can be used for killing gram-negative bacteria and gram-positive bacteria. The antibacterial peptide prepared by the invention can be developed into anti-bacterial infection drugs or feed additives, and is applied to prevention and treatment of bacterial diseases of aquaculture animals.

Disclosed herein are methods to assay and identify detection reagents for chromatin elements, including recombinant fusion protein detection reagents comprising two or more binding domains for chromatin elements. Improved genomic mapping assays using the identified recombinant proteins are also disclosed. Recombinant fusion proteins optimized by the disclosed methods are also provided.

Provided is a polypeptide specifically binding to a transferrin receptor, which is suitable for the technical field of molecular biology. The polypeptide comprises the general formula H1-H2-E1-E2-H3-H4-E3, wherein the H1, H2, H3 and H4 each independently comprise an α-helical domain; and the E1, E2 and E3 each independently comprise a β-sheet domain. The polypeptide binds to hTfR1 while allowing transferrin to bind to hTfR1. The polypeptide can bind to and deliver an oligonucleotide. Further provided are a recombinant nucleic acid encoding the polypeptide, an expression vector comprising the recombinant nucleic acid, a recombinant host cell comprising the polypeptide, the nucleic acid and / or the expression vector, a conjugate of the polypeptide and an oligonucleotide, and a corresponding pharmaceutical composition. The polypeptide can bind to hTfR1 with a high specificity, so as to achieve accurate delivery of the oligonucleotide, and thereby providing a real basis for the treatment of gene mutation-related diseases.

Various embodiments relate generally to the technical field of enzyme technology and specifically relates to enzymes having depolymerisation activity and to nucleic acids encoding those as well as method of the manufacture of said enzymes. In particular, the enzymes are nylon hydrolases. Further encompassed are methods and uses of these enzymes.

The invention provides a composition for preventing and treating infection of mycoplasma hyorhinis. The composition takes a C-terminus of DnaK and an N-terminus-XylF fusion protein of P72 as an active ingredient. The invention further provides an expression vector and a method for producing the fusion protein.