126results about "Polypeptide with GST-tag" patented technology

The present application discloses a fusion protein comprising a varicella zoster virus protein and a ferritin, a nucleic acid encoding the fusion protein, a pharmaceutical composition comprising the fusion protein, a method of preparing the fusion protein, and a use of the fusion protein.

The invention discloses a method for preparing a simeglutide precursor peptide through intracellular enzyme digestion. The method comprises the following steps: (1) connecting a dissolution-promoting tag GST and the simeglutide precursor peptide together through protease specific recognition of amino acid sites to form a fusion protein; (2) integrating the coding nucleotide sequence of the fusion protein, the coding nucleotide sequence of KEN2 protease and the coding nucleotide sequence of carboxypeptidase B into the same expression vector to form a recombinant vector; (3) transforming the recombinant vector into engineering bacteria and carrying out fermentation production; and (4) centrifuging a fermentation product, collecting thalli, and purifying to obtain the precursor peptide powder of the semeglutide. The whole operation process is simpler, more convenient and smoother, tedious recovery steps are omitted, and the preparation and recovery difficulty of the precursor peptide of the semeglutide is remarkably reduced.

Provided is a low molecular weight collagen with a 164.88° triple helix structure. Relates to a small molecule collagen, a preparation method and use thereof. The collagen of the present application comprises an amino acid sequence as shown in SEQ ID NO:1 or a variant amino acid sequence after mutation of the amino acid sequence, and the variant amino acid sequence retains the function of the amino acid sequence as shown in SEQ ID NO:1. The amino acid sequence of the collagen of the present application is shorter and has better transdermal absorption performance.

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.

Peptides that specifically bind erythrocytes are described. These are provided as peptidic ligands having sequences that specifically bind, or as antibodies or fragments thereof that provide specific binding, to erythrocytes. The peptides may be prepared as molecular fusions with therapeutic agents, tolerizing antigens, or targeting peptides. Immunotolerance may be created by use of the fusions and choice of an antigen on a substance for which tolerance is desired.

The invention relates to the technical field of organelle separation, and provides a combination of ATF6 alpha N protein and peroxisome, application of the ATF6 alpha N protein in separation and purification of the peroxisome and a method for separating and purifying the peroxisome. The ATF6 alpha N protein is combined with membrane protein PMP70 and / or ABCD1 of the peroxisome; the method comprises the following steps: combining tagne-ATF6 alpha N fusion protein with a medium to obtain a fusion protein-medium compound; adding a roughly separated peroxisome sample into the fusion protein-medium compound, so as to obtain a fusion protein-medium-peroxisome compound; and adding an elution buffer solution, and collecting the eluent to obtain the peroxisome combined with the fusion protein. The method is easy and convenient to operate, complex instruments and equipment are not needed, the peroxisome can be separated from cells and tissues, and the obtained peroxisome is high in purity and can be used for conventional experiments such as subsequent protein spectrum analysis and flow analysis.

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 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 present invention relates to a method for producing a plant or isolated plant cells for producing collagen, a nucleic acid construct used therein, and a method for producing collagen using the plant or isolated plant cells.

The invention discloses application of RNA (Ribonucleic Acid) binding protein CsdA in regulation and control of secondary metabolites of fungi. The invention provides application of RNA (Ribonucleic Acid) binding protein CsdA or related biological materials thereof in regulation and control of fungus secondary metabolites. The RNA binding protein CsdA is a protein of which the amino acid sequence is as shown in SEQ ID No. 3 or SEQ ID No. 2 or a homologous protein of the protein of which the amino acid sequence is as shown in SEQ ID No. 3 or SEQ ID No. 2. The invention is helpful for excavating natural products with medicinal activity in fungi, is helpful for developing new anti-fungal infection drug targets, and reduces the probability of fungal infection.

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.

The present disclosure relates, in some aspects, to recombinant polymerases, variant PumA1 polymerases, and compositions thereof. Also provided herein are methods of using the recombinant polymerases and / or variant PumA1 polymerases for nucleic acid amplification (e.g., rolling circle amplification). In some aspects, the compositions and methods disclosed herein provide more robust amplification (e.g., RCA) reactions for improved in vitro and in situ analysis.

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 relates to the technical field of biology. Disclosed are recombinant enterobacter and a use thereof in degradation of tyrosine and phenylalanine. According to the present invention, enzyme molecules in nature that have both phenylalanine and tyrosine degradation activities are collected by means of gene search, and the phenylalanine and tyrosine degradation activities of the enzyme molecules are tested in vitro; enzyme molecules having excellent activities are obtained by screening; the selected enzyme molecules are transferred into enterobacter, engineered bacteria highly expressing the enzyme molecules and having the ability to efficiently degrade phenylalanine and tyrosine are further selected, and then protein tags are introduced to improve the degradation activities. Finally, an optimal structure is selected by means of gastrointestinal fluid tolerance experiments, an engineered bacterium having degradation activities far higher than the existing degradation activities is obtained, and the engineered bacterium is expected to be used for preparing a drug for treating phenylketonuria and tyrosinase.

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 present invention provides a composition for preventing and treating a mycoplasma hyorhinis infection. The composition uses the N-terminal-XylF fusion protein of C-terminal-P72 of DnaK as an active ingredient. The present invention further provides an expression vector and a method required for producing the fusion 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 belongs to the technical field of biology, and particularly relates to a bovine viral diarrhea virus NS2 truncated protein and application thereof in BVDV antibody detection. The invention firstly provides a truncated protein of a BVDV NS2 protein, the truncated protein is subjected to soluble expression after being fused with glutathione S transferase (GST) and horse radish peroxidase (HRP) respectively, has better reactogenicity with positive serum of a BVDV antibody, does not react with negative serum of the BVDV antibody, and can be used for detecting the BVDV antibody; secondly, a fusion protein of GST and the truncated protein is taken as a coating antigen, a fusion protein of HRP and the truncated protein is taken as a detection antigen, a double-antigen sandwich ELISA method for BVDV antibody detection is constructed, and the method has the technical advantages of high sensitivity, good specificity, simplicity and convenience in operation, easiness in popularization, low cost and the like, and can be applied to screening of BVDV antibodies in cattle.

A PD-L1 mutant having improved binding affinity for PD-1 is disclosed. A method for preparing the PD-L1 variant and a method for screening the PD-L1 variant are also disclosed. The PD-L1 variant produced by substituting some amino acids in the sequence of wild-type PD-L1 with other optimal amino acids, achieving greatly improved affinity for PD-1. In addition, the possibility of immunogenicity can be reduced by the smallest possible number of the mutation sites.

An scFv-Fc dimer binds and neutralizes TGFβ1 selectively and with high affinity and avidity. The scFv region may comprise the same VH and VL domains or CDR regions as metelimumab. The unique combination of their smaller size, high selectivity, potency against TGFβ1, and long in vivo half-life makes the scFv-Fc dimers ideal candidates for therapeutic applications.