59results about "Polypeptide with GST-tag" patented technology

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.

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.

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 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 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.

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.

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.

A method is described for the rapid clearance of carbon monoxide (CO) from CO-bound hemoglobin, myoglobin, and cytochrome c oxidase in subjects with CO poisoning. The disclosed therapeutic method involves the use of rationally designed and engineered regulator of CO metabolism (RcoM) proteins and pharmaceutical compositions thereof, which scavenge CO from poisoned tissues. The recombinant RcoM composition is infused into the blood, where it rapidly sequesters CO and limits the toxic effects of CO on cellular respiration, oxygen transport, and oxygen utilization.

Provided are a fusion protein including glutathione-S-transferase and a protein having binding affinity for a target cell or a target protein, and use thereof as a drug delivery carrier and a pharmaceutical composition. The fusion protein and the drug delivery carrier including the same according to an aspect may sustain an in vivo residence time, and may also have improved target cell-targeting ability, and thus may be effectively delivered to target cells. Accordingly, it may be usefully applied to a targeted therapeutic agent.

To provide RcoM protein-based carbon monoxide scavengers and preparations for treating carbon monoxide poisoning.SOLUTION: Disclosed herein is a method for rapid elimination of carbon monoxide (CO) from CO-bound hemoglobin, myoglobin and cytochrome C oxidase in a subject with CO poisoning. The disclosed therapeutic method comprises use of a rationally designed and modified regulatory factor of CO metabolism (RcoM) protein and pharmaceutical compositions thereof, which scavenge carbon monoxide from poisoned tissue. The recombinant RcoM composition is infused into blood to rapidly sequester carbon monoxide and limit the toxic effects of carbon monoxide on cellular respiration, oxygen transport and oxygen utilization.SELECTED DRAWING: None

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 relates to a recombinant chondroitin-6-O-sulfotransferase mutant and application of the recombinant chondroitin-6-O-sulfotransferase mutant in synthesis of chondroitin sulfate C, and belongs to the technical field of bioengineering. According to the invention, a dissolution promoting label is fused at the N end of a chondroitin-6-O-sulfotransferase mutant, so that the efficient soluble expression of the enzyme is successfully realized, and the proportion of soluble components is as high as 90%. The mutant HsC6ST-delta loop has the advantages that by carrying out amino acid sequence truncation on chondroitin-6-O-sulfotransferase, the constructed mutant HsC6ST-delta loop has higher chondroitin-6-O-sulfonation activity compared with HsC6ST chondroitin-6-O-sulfonation, the sulfonation rate reaches 78.4% after reaction is carried out for 12 h under the conditions that the temperature is 40 DEG C and the pH is 8.0, and the Tm and half-life period are increased by 9 DEG C and 1.54 h compared with those of an original enzyme.

This invention relates to the terpene synthase AaTPS48 from Artemisia argyi, its encoding gene, and its applications. It provides an important foundation for utilizing genetic engineering techniques to increase the content of the active components myrcene and (trans)-β-farnesene in Artemisia argyi, or to directly produce myrcene and (trans)-β-farnesene. The technical solution is that the terpene synthase AaTPS48 is a protein of the following a) or b) or c): a) the amino acid sequence of the protein shown in SEQ ID No. 1; b) a fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of the protein shown in SEQ ID No. 1; c) a protein with the same function obtained by substituting and / or deleting and / or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID No. 1. The AaTPS48 protein of this invention can catalyze both GPP to form myrcene and FPP to form (trans)-β-farnesene, playing an important role in the biosynthesis of myrcene, (trans)-β-farnesene, and other terpene compounds in Artemisia argyi.

The presently claimed subject matter concerns methods and kits for identifying agents that reduce binding of a clostridial neurotoxin to synaptic vesicle glycoprotein 2 (SV2).

The invention relates to disulfide-rich peptides derived from spider venom and their use, particularly as neuroprotective agents. The invention also relates to nucleic acid molecules encoding the peptides as well as constructs and host cells comprising those nucleic acid molecules.

The application discloses a kind of peptidoglycan recognition protein-D, preparation method and application, belong to biological medicine technical field.The application obtains natural peptidoglycan recognition protein-D from tussah by protein separation and purification technology, realizes the expression of peptidoglycan recognition protein-D and its derivative or analogue or partial fragment gene in host cell after resolving its primary structure (gene and protein) using genetic engineering technology, and the antibody of purified recombinant peptidoglycan recognition protein-D and its derivative or analogue or partial fragment is obtained by immunizing animal.The natural, recombinant peptidoglycan recognition protein-D and its derivative or analogue or partial fragment and its antibody of the application can be widely used for prevention, detection diagnosis, treatment and other biological medicine fields of microorganism, microorganism related molecular pattern.

The invention discloses a vector, a construction method thereof and application of the vector to FGF (fibroblast growth factor) obtained by expression based on a nicotiana benthamiana chassis, and belongs to the field of basic fibroblast growth factor production. The technical scheme of the invention is as follows: the biomarker comprises a carrier pJL-TRBOGZ082 or a carrier pJL-TRBGZ125; the vector pJL-TRBOGZ082 comprises a 6Xhis-FGF (Fibroblast Growth Factor) sequence as shown in SEQ ID No.1 (Sequence Identifier Number 1); the vector pJL-TRBGZ125 contains a GST (Glutathione S Transferase)-FGF (Fibroblast Growth Factor)-6 Xhis sequence as shown in SEQ ID No. 2. The method is applied to the production of the basic fibroblast growth factor, and solves the technical problems that the related research on the production of the bFGF by utilizing a tobacco transient expression technology at present is not sufficient, and particularly, the key links such as expression vector optimization, permeation infection condition control and protein purification process are still lack of system optimization. The invention provides an efficient and low-cost FGF (Fibroblast Growth Factor) production vector based on a tobacco transient expression system, a construction method of the vector and an application of the vector in obtaining the FGF based on the expression of a nicotiana benthamiana chassis.

The present application provides a polypeptide specifically binding to PDL1 and its use. Specifically, the present application relates to a polypeptide or an active fragment thereof capable of specifically binding to PDL1, and a fusion protein comprising the polypeptide or the active fragment thereof. The present application also provides a CAR-T cell capable of expressing the fusion protein, and the CAR-T cell is used to specifically kill tumor cells, such as lung cancer, multiple myeloma or acute myeloid leukemia. The CAR-T of the present application can be used as a therapeutic drug for tumor diseases, and provides a new method for the prevention and treatment of tumors.

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 invention concerns a novel fusion polypeptide comprising a polypeptide having an N- glycosidase activity fused to an antioxidant moiety, wherein the polypeptide having an N-glycosidase activity is a polypeptide from a bacterium belonging to the Acidobacteria phylum, preferably to the Acidobacteriia class. The present invention also concerns the nucleic acid molecule, vector, and host cell encoding the fusion polypeptide, as well as compositions and kits thereof. The present invention also provides methods for purifying and / or for producing the fusion polypeptide, as well as in vitro use of the fusion polypeptide, or the nucleic acid molecule, vector, or host cell encoding the fusion polypeptide, or compositions thereof.

The present invention relates to a XIAP protein variant or a fusion protein comprising same, and use thereof. The XIAP protein variant according to one embodiment has an effect functionally equivalent to that of a wild-type XIAP protein, promotes ubiquitination of receptor-interacting serine / threonine-protein kinase 2 (RIP2) so as to activate NOD signaling, and has increased residual time and levels in cells relative to the wild-type XIAP protein, and thus can be effectively used in the treatment of inflammatory diseases, particularly diseases caused by NOD signaling defects.

The cryo-electron microscope sample preparation kit contains an amphipathic protein and a protein crosslinking agent, or an expression vector containing a nucleic acid sequence encoding the amphipathic protein. The kit also includes an ice thickness control molecule to which an amphipathic protein is attached. The amphipathic protein may have an amphipathic α-helix structure and may be either an antifreeze protein or a type I antifreeze protein.

Provided herein are multivalent protein scaffolds that are useful as therapeutic agents and are useful for identifying new therapeutic compounds.The present invention also relates to multi-domain polypeptide constructs that have multiple binding domains and a structural domain.Also provided herein are methods for identifying new candidate therapeutic agents using the provided multivalent protein scaffolds, and the new therapeutic agents identified thereby.

The invention provides a designable fusion protein for expression of small-molecule collagen peptide as well as a recombinant vector and a recombinant expression method thereof, and belongs to the technical field of recombinant protein expression. The fusion protein for collagen peptide expression comprises the following connected fragments: a soluble tag, a purification tag, a connecting peptide, a protease digestion recognition sequence and a designed target collagen peptide, the amino acid sequence of the connecting peptide is as shown in SEQ ID NO: 1. And the recombinant fusion protein is subjected to purification, enzyme digestion treatment, re-purification, refining and separation to obtain a peptide fragment with a target molecular weight. Experiments show that the method can obtain the small molecular collagen peptide with small molecular weight, designable sequence and yield reaching the level of gram per liter on the premise of not depending on chemical synthesis and subsequent random enzymolysis, and the scheme is simple and easy to implement, stable in preparation process, high in purity and suitable for industrial large-scale production.