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36 results about "TEV protease" patented technology
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TEV protease (EC 3.4.22.44, Tobacco Etch Virus nuclear-inclusion-a endopeptidase) is a highly sequence-specific cysteine protease from Tobacco Etch Virus (TEV). It is a member of the PA clan of chymotrypsin-like proteases. Due to its high sequence specificity it is frequently used for the controlled cleavage of fusion proteins in vitro and in vivo.
Recombinant I-type humanized collagen polypeptide as well as preparation method and application thereof
ActiveCN113621052AIncrease productionPromote cell adhesionCosmetic preparationsBacteriaCell adhesionProteinogenic amino acid
The invention discloses a recombinant I-type humanized collagen polypeptide as well as a preparation method and application thereof. The recombinant I-type humanized collagen polypeptide provided by the invention comprises n repeats of a sequence shown in SEQ ID No. 1, wherein n is an integer greater than or equal to 1, and when n is an integer greater than or equal to 2, the repeat sequences are directly connected; and optionally, the N tail end of the recombinant I-type humanized collagen polypeptide comprises an amino acid sequence which can be excised by TEV protease. The recombinant I-type humanized collagen polypeptide provided by the invention has the activity of promoting cell adhesion, the amino acid sequence of the recombinant protein is selected from a natural collagen amino acid sequence, and the recombinant protein does not generate an immune response when being applied to a human body; and moreover, the preparation method is simple, and high-yield collagen can be obtained at low cost.
Owner:SHANXI JINBO BIO PHARMA CO LTD
Producing a Target Protein Using Intramolecular Cleavage by TEV Protease
A cis-TEVP fusion protein including a TEVP protease, a TEVP cleavage site and a target protein provide a platform for expression of the target protein. A trans-TEVP fusion protein including a TEVP cleavage site and a target protein, the amino-terminal portion of the target protein adjacent to the C-terminal portion of the TEVP cleavage site, the amino acidic residue in position P2 of the TEVP cleavage site being a Valine also produces the target protein by the same process. A cis-TEVP fusion protein system comprising the first fusion protein and a suitable host cell; a trans-TEVP fusion protein system comprising the second fusion protein and a suitable host cell; associated methods to produce target proteins, and kits of parts are also disclosed herein.
Owner:ACAD SINIC
Processing of a modified foot-and-mouth disease virus p1 polypeptide by an alternative protease
ActiveUS20180200359A1SsRNA viruses positive-senseViral antigen ingredientsProteinase activityVirus-like particle
Polynucleotide constructs that express an engineered foot-and-mouth disease (FMDV) P1 precursor protein and a non-FMDV TEV protease and methods for safe and efficient recombinant production of FMDV antigens and immunogens. Recombinant production of FMDV antigens avoids the need to culture highly-infectious FMDV, while conventional culture methods for producing FMDV antigens rely on the native FMDV 3C protease which exerts toxic effects on host cells. The inventors have developed a new system that efficiently and safely processes FMDV P1 precursor without the FMDV 3C protease, thus avoiding the toxic effects associated with use of the 3C protease. The invention is also directed to the FMDV antigens and virus-like particles produced by this system as well as to FMDV vaccines, diagnostics and other biologics.
Owner:THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SEC OF HOMELAND SECURITY
Method for engineering proteases and protein kinases
ActiveUS8945855B2High activityEfficient identificationHydrolasesMicroorganismsEngineering proteinYeast display
Provided are methods for protein engineering, such as engineering proteases or kinases. The methods may utilize yeast display and / or ER sequestration of proteins or substrates. In some aspects, TEV proteases with altered substrate specificity, potency, and / or efficiency are provided.
Owner:RES DEVMENT FOUND
Processing of a modified foot-and-mouth disease virus P1 polypeptide by an alternative protease
Polynucleotide constructs that express an engineered foot-and-mouth disease (FMDV) P1 precursor protein and a non-FMDV TEV protease and methods for safe and efficient recombinant production of FMDV antigens and immunogens. Recombinant production of FMDV antigens avoids the need to culture highly-infectious FMDV, while conventional culture methods for producing FMDV antigens rely on the native FMDV 3C protease which exerts toxic effects on host cells. The inventors have developed a new system that efficiently and safely processes FMDV P1 precursor without the FMDV 3C protease, thus avoiding the toxic effects associated with use of the 3C protease. The invention is also directed to the FMDV antigens and virus-like particles produced by this system as well as to FMDV vaccines, diagnostics and other biologics.
Owner:THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SEC OF HOMELAND SECURITY
Fusion expression method of AEP cyclase in Escherichia coli, method for identifying cyclization capacity of AEP cyclase and application of APE cyclase
ActiveCN110331157ABroad-spectrumHigh expressionMicrobiological testing/measurementNucleic acid vectorEscherichia coliCyclase
The invention belongs to the technical field of gene engineering and particularly relates to a fusion expression method of AEP cyclase in Escherichia coli, a method for identifying cyclization capacity of the AEP cyclase and application of the APE cyclase. Fusion expression is performed in the Escherichia coli by different fusion labels, then, the AEP cyclase with higher expression is obtained after the fusion labels are removed, and the expression of the AEP cyclase is 0.36 plus / minus 0.05 mg / mL, which is higher than 1.8 mg / L reported in original literature. The new method is adopted to verify the cyclization capacity of the AEP cyclase, protein which can be better observed in SDS-PAGE and has cyclization probability is selected by constructing a serial substrate, a serial substrate containing a substrate sequence of TEV protease is constructed, whether cyclization is performed according to change of the size of the substrate or the number of stripes after protease cutting, and the method is simple to operate, convenient and high in practicability.
Owner:湖北凯利特生物科技有限公司
TEV protease mutant, gene, biomaterial, preparation method, reagent, or reagent kit and application
The invention provides a TEV protease mutant, a gene, a biomaterial, a preparation method, a reagent, or a reagent kit and application, and relates to the technical field of genetic engineering. Compared with a protease mutant having an amino acid sequence shown as SEQ ID NO.1, the TEV protease mutant is mutated into at least seven amino acid residues, for example 17th site is mutated into S fromT, the 56th site is mutated into V from L, the 68th site is mutated into D from N, the 77th site is mutated into V from I, the 135th site is mutated into G from S, the 219th site is mutated into V from S, and the 233th site is mutated into H from Q. The TEV protease mutant has favorable enzymatic activity, stability and specificity. The gene, the biomaterial and the preparation method of the TEV protease mutant, or the reagent or the reagent kit containing the TEV protease mutant, the application of the technical scheme, and the TEV protease mutant are all based on the same invention conception.
Owner:生工生物工程(上海)股份有限公司
Expression method of AEP cyclase in pichia pastoris and application thereof
ActiveCN110387366AIncreased cyclization activityImprove stabilityNucleic acid vectorVector-based foreign material introductionCyclasePichia pastoris
The invention belongs to the technical field of genetic engineering, and particularly relates to an expression method of AEP cyclase in pichia pastoris and application thereof. The AEP cyclase is expressed in pichia pastoris, the gene tandem copy number of the AEP cyclase reaches up to 5 copies by a gene tandem way, and the expression amount of the AEP cyclase is highest up to 0.89+ / -0.03 mg / mL when the gene copy number reaches 3. The invention verifies the cyclization capacity of the AEP cyclase in a novel mode, constructs a tandem substrate, selects a protein as the substrate which can be better observed in SDS-PAGE and has the cyclization possibility, constructs a tandem substrate containing the substrate sequence of the TEV protease; and identifies whether the AEP cyclase is cyclized or not according to the size change of the substrate after the protease is cut or the number of bands. The method is simple and convenient to operate and strong in practicability.
Owner:湖北凯利特生物科技有限公司
Processing engineered fmdv p1 polypeptide using an alternative tev protease
ActiveUS20190211321A1SsRNA viruses positive-senseViral antigen ingredientsDiseaseVirus-like particle
Polynucleotide constructs that express an engineered foot-and-mouth disease (FMDV) P1 precursor protein and a non-FMDV TEV protease and methods for safe and efficient recombinant production of FMDV antigens and immunogens. Recombinant production of FMDV antigens avoids the need to culture highly-infectious FMDV, while conventional culture methods for producing FMDV antigens rely on the native FMDV 3C protease which exerts toxic effects on host cells. The inventors have developed a new system that efficiently and safely processes FMDV P1 precursor without the FMDV 3C protease, thus avoiding the toxic effects associated with use of the 3C protease. The invention is also directed to the FMDV antigens and virus-like particles produced by this system as well as to FMDV vaccines, diagnostics and other biologics.
Owner:THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SEC OF HOMELAND SECURITY
TEV protease variant, fusion protein, preparation method and uses thereof
ActiveCN111019926AUnique access to performanceHigh activityAntibody mimetics/scaffoldsCorticotropinBiochemistryProtease
The invention provides a TEV protease variant, a fusion protein, a preparation method and uses thereof, particularly a screened TEV protease variant with unique properties, and a fusion protein. According to the invention, polypeptides can be rapidly and efficiently prepared through TEV protease variant and polypeptide fusion expression so as to solve the problems existing in the current polypeptide recombinant production process.
Owner:SHANGRAO CONCORD PHARMA CO LTD
Method for recombining, expressing and producing human thymosin in yeast
ActiveCN102925470AEnsure product quality consistencyLow costHybrid peptidesVector-based foreign material introductionNicotiana tabacumRestriction enzyme digestion
The invention provides a method for massively producing human thymosin with low cost. The method is used for implementing restriction enzyme digestion to obtain a fusion gene sequence with a structure as follows: A-X-C, wherein A is a nucleotide sequence of site (1-150)-(1-372) amino acids at the N- end of coded mature human serum albumin; X is the nucleotide sequence of connecting peptide with enterokinase or tobacco etch virus (TEV) protease cutting site contained in a code; and C is a human thymosin gene. The method comprises steps as follows: connecting the fusion gene sequence to an expression carrier; transforming and introducing the expression carrier into saccharomycetes; carrying out induction expression to obtain soluble human serum albumin-thymosin fusion protein; then adding the TEV protease for cutting; and separating and purifying to obtain the recombined human thymosin. The human thymosin production method provided by the invention has the advantages that consistency of quality of products can be ensured, no limitation is generated from sources of raw materials, cost is low, an expression index is high, and mass production can be achieved and the like.
Owner:冯鹏波
Method for engineering proteases and protein kinases
ActiveUS20140017764A1High activityEfficient identificationHydrolasesMicroorganismsEngineering proteinYeast display
Provided are methods for protein engineering, such as engineering proteases or kinases. The methods may utilize yeast display and / or ER sequestration of proteins or substrates. In some aspects, TEV proteases with altered substrate specificity, potency, and / or efficiency are provided.
Owner:RES DEVMENT FOUND
Method for producing recombinant TEV protease in escherichia coli
InactiveCN108774635AReduce process timeReduce purification timeHydrolasesVector-based foreign material introductionEscherichia coliFiltration
The invention discloses a method for producing recombinant TEVprotease in escherichia coli. The method comprises the following steps: S1) construction of PET28aTEV protease expression plasmid; S2) inducing expression and identification of TEV protease; and S3) purification and identification of TEV protease. The invention adopts the escherichia coli expression system to realize the expression of recombinant TEV protease in escherichia coli by designing an expression vector and then adopts affinity chromatography and Superdex 75 gel filtration chromatography to purify the target protease, thusgreatly shortening the purification process and time of the target protease, improving the purity, yield and activity of the protease, saving experimental steps and reducing cost. The method providedby the invention lays a foundation for the development and production of other protein tool enzymes.
Owner:通用生物(安徽)股份有限公司
Method of shearing fusion protein by escherichia coli intracellular protease
InactiveCN103993030AActivity does not affectRetain activityFermentationVector-based foreign material introductionProtein targetThiogalactosides
The invention relates to a method of separating a target protein by shearing a fusion protein by escherichia coli intracellular protease. Used host bacteria is obtained by integrating a TEV (tobacco etch virus) protease gene driven by a strong promoter T7 to escherichia coli expression host bacteria BL21(DE3) by virtue of a recombinant engineering method. An expression vector is obtained by cloning maltose-binding protein and stuffer fragment to the expression vector pET30(+). A target gene not containing a terminator replaces the stuff segment to obtain a target gene fusion expression vector, and the tail end of the fusion protein contains 6 histidines of the vector. After the expression vector is transformed to the host bacteria, under induction of isopropyl-beta-D-thiogalactoside, the fusion protein and the TEV protease based on a chromosome are expressed at the same time; the TEV is acted on an enzyme cutting site between the maltose-binding protein and the target protein for separating the target protein. According to the intracellular shearing method, steps of fusion protein separating, TEV enzyme digesting, and the like are omitted.
Owner:NANJING NORMAL UNIVERSITY
Novel method for preparing recombinant exenatide or derivative thereof
InactiveCN104894196AEasy to detectHas anticoagulant activityHormone peptidesFermentationFusion Protein ExpressionEnzyme digestion
The invention builds up a novel method for preparing recombinant exenatide or derivative thereof. According to the method, hirudin (molecular weight is 7Kd) serves as a fusion partner, exenatide or derivative thereof is spiced on the downstream of the hirudin for fusion expression, a connecting peptide is arranged between the hirudin fusion partner and the exenatide or derivative thereof and comprises a TEV protease (tobacco etching virus protease) identification cutting sequence ENLYFQH. The exenatide or derivative thereof with a natural N-terminal and biological activity is released through TEV enzyme digestion after fusion protein expression. The novel method has other advantages that (1) the hirudin fusion partner is small, so the ratio of the exenatide or derivative thereof in fusion protein is effectively increased; (2) hirudin fusion protein has anticoagulation activity, thus facilitating real-time detection and tracing; (3) fusion protein can be adsorbed by virtue of cheap macroporpous resin; and (4) hirudin fusion partner can enhance stability of exenatide or derivative thereof.
Owner:CHINA PHARM UNIV
Method for screening protease variant and obtained protease variant
PendingUS20220025355A1Weak cleavage activityEfficient cuttingCorticotropinFusion with protease siteEnzyme variantBacteriophage
The present invention relates to a method for screening a protease variant and the obtained protease variant, and the protease variant has a special performance. The method for screening the protease variant comprises the following steps: (1) constructing a protease random mutation library, optimizing a protease random mutation virus library, and optimizing a protease random mutation phage library; (2) for the protease random mutation library, optimizing the protease random mutation virus library, and optimizing the protease random mutation phage library and screening a protease variant having weak protease enzyme-cutting activity in a host or a similar condition and having protease enzyme-cutting activity in an in vitro denaturation condition; and (3) the step of optionally characterizing the protease variant, wherein the protease optimizes TEV protease or enterokinase. The protease variant in the present invention facilitates industrial production of protein.
Owner:SHANGRAO CONCORD PHARMA CO LTD
Efficient-expression recombinant TEV enzyme with high activity and stability as well as preparation method, determination method and application of recombinant TEV enzyme
Owner:无锡佰翱得生物科学股份有限公司
Method for efficiently expressing hypersensitive protein by using T4 phage display technology
PendingCN110878320AFast preparation methodBiocidePlant growth regulatorsEscherichia coliT4 bacteriophage
The invention relates to a method for efficiently expressing hypersensitive protein by using a T4 phage display technology. The method comprises the following steps: (1) sequentially connecting an encoding gene of the hypersensitive protein with TEV protease and a T4 bacteriophage Hoc protein gene segment to prepare a fusion recombinant protein encoding gene, wherein the encoding region of the hypersensitive protein is located at the upstream of a TEV-Hoc fusion protein gene and having consistent expression cassettes; (2) transferring a coding gene of the fusion recombinant protein into a pUC18 plasmid; (3) transferring the plasmid into an escherichia coli host; (4) infecting the Escherichia coli host by using Hoc-T4 bacteriophage deleted by Hoc protein, and culturing the host to express and display the T4 bacteriophage of the fusion recombinant protein; (5) separating the T4 bacteriophage showing the fusion recombinant protein; and (6) separating the fusion recombinant protein from the T4 bacteriophage, and carrying out TEV protease digestion to obtain the hypersensitive protein.
Owner:湖北微生元生物科技有限公司
Method for high-efficiency expression of all subtype mature proteins of IL-37 by utilizing tobaccos
InactiveCN106434745AKeep alive functionInhibition of secretionPolypeptide with localisation/targeting motifFusion with protease siteDiseaseBiotechnology
The invention discloses a method for high-efficiency expression of all subtype mature proteins of IL-37 by utilizing tobaccos. The method comprises the following steps of: inserting coding sequences of TEV protease and enzyme cutting sites thereof between coding sequences of signal peptide and mature IL-37 proteins to form an IL-37-TEV fusion gene; according to the preference of tobacco codons, under the condition of guaranteeing no change of the amino acid sequence, optimizing the sequence of the fusion gene; connecting 5'-end upstream of the constructed IL-37-TEV fusion gene with a common promoter including other regulating areas, and connecting the 3'-end downstream with a transcription terminator; integrating a constructed recombinant expression vector onto cell chromosomes of the tobacco to culture a transgenic tobacco plant with high-efficiency stable expression of IL-37 and injecting the constructed recombinant expression vector to the tobacco leaves to instantaneously express target proteins. The method disclosed by the invention has the advantages that by construction of the plant expression vector, the mature proteins of IL-37 with high activity are expressed in the plants, and the obtained product has important application value for treatment of numerous inflammatory diseases and early detection and diagnosis of the inflammatory diseases.
Owner:马生武 +4
A method for high-throughput screening of single-domain antibodies using ispla and its application
ActiveCN113528569BNo preparation involvedHigh affinityAntibody mimetics/scaffoldsLibrary screeningAntigenPseudomonas aeruginosa exotoxin A
The invention discloses a method for high-throughput screening of single-domain antibodies using isPLA. The steps are as follows: constructing an sdAb library containing 21 random amino acid sequences in the CDR3 region, and fusing a 3×Flag tag to the C-terminus of the sequence; co-transfection Cells, fixed cells for isPLA; followed by sorting, amplification, and recombination, and after multiple rounds of screening and sequencing, constructs that sequentially contain glutathione S-transferase GST, tobacco mosaic virus TEV protease cleavage site, and recognize SQSTM1. The sdAb sequence, the translocation domain of Pseudomonas exotoxin ETA and the 3×Flag-tagged recombinant protein were expressed in E. coli, followed by purification and enzyme digestion. The method can screen sdAbs that specifically recognize different antigenic determinants from the constructed high-capacity sdAb library, and has low cost and high efficiency. No preparation of animal samples or hybridomas is involved.
Owner:TIANJIN MEDICAL UNIV
tev protease mutant, gene, biological material, preparation method, reagent or kit and application
The invention provides a TEV protease mutant, a gene, a biomaterial, a preparation method, a reagent, or a reagent kit and application, and relates to the technical field of genetic engineering. Compared with a protease mutant having an amino acid sequence shown as SEQ ID NO.1, the TEV protease mutant is mutated into at least seven amino acid residues, for example 17th site is mutated into S fromT, the 56th site is mutated into V from L, the 68th site is mutated into D from N, the 77th site is mutated into V from I, the 135th site is mutated into G from S, the 219th site is mutated into V from S, and the 233th site is mutated into H from Q. The TEV protease mutant has favorable enzymatic activity, stability and specificity. The gene, the biomaterial and the preparation method of the TEV protease mutant, or the reagent or the reagent kit containing the TEV protease mutant, the application of the technical scheme, and the TEV protease mutant are all based on the same invention conception.
Owner:生工生物工程(上海)股份有限公司
Method for preparing high purity cryptoprotein
InactiveCN109666686ASimple purification processEasy accessPlant growth regulatorsBiocideEscherichia coliProtein target
The invention provides a method for preparing high-purity cryptoprotein. The method comprises the steps of (1) sequentially connecting a coding gene of a target protein with a TEV protease and a T4 phage Soc protein gene fragment to prepare a fusion recombinant protein coding gene; (2) transferring the gene encoding the fusion recombinant protein into a plasmid; (3) transferring the plasmid into abacterium coli host; (4) infecting the bacterium coli host with a Soc-deficient Soc-T4 phage, and cultivating the host to express the T4 phage displaying the fusion recombinant protein; (5) isolatingthe T4 phage displaying the fusion recombinant protein; (6) isolating the fusion recombinant protein from the T4 phage, and obtaining the target protein by TEV protease digestion. The method can obtain the high-purity cryptoprotein easily and quickly, an obtained cryptoprotein solution can be directly used as an agricultural antibacterial bacteriostatic agent to achieve the effect of promoting plant growth, and the method has a good application prospect.
Owner:EZHOU INST OF IND TECH HUAZHONG UNIV OF SCI & TECH +1
Method for efficiently expressing hypersensitive protein by using T4 phage display technology
PendingCN110885847AFast preparation methodBiocideAntibody mimetics/scaffoldsEscherichia coliT4 bacteriophage
The invention relates to a method for efficiently expressing hypersensitive protein by using a T4 phage display technology. The method comprises the following steps: (1) sequentially connecting an encoding gene of the hypersensitive protein with TEV protease and a T4 bacteriophage Soc protein gene segment to prepare a fusion recombinant protein encoding gene, wherein a encoding region of the hypersensitive protein is located at the upstream of a TEV-Soc fusion protein gene and having consistent expression cassettes; (2) transferring the coding gene of the fusion recombinant protein into a pUC18 plasmid; (3) transferring the plasmid into an escherichia coli host; (4) infecting the escherichia coli host by using Soc-T4 bacteriophage without Soc protein, and culturing the T4 bacteriophage ofwhich the host expresses and displays the fusion recombinant protein; (5) separating the T4 bacteriophage showing the fusion recombinant protein; and (6) separating the fusion recombinant protein fromthe T4 bacteriophage, and carrying out TEV protease digestion to obtain the hypersensitive protein.
Owner:湖北微生元生物科技有限公司
A kind of expression method and application of aep cyclase in Pichia pastoris
ActiveCN110387366BIncreased cyclization activityImprove stabilityNucleic acid vectorVector-based foreign material introductionCyclaseGenetic engineering
The invention belongs to the technical field of genetic engineering, and in particular relates to an expression method and application of AEP cyclase in Pichia pastoris. In the present invention, by expressing in Pichia pastoris, the number of copies of the AEP cyclase gene in series can reach up to 5 copies through the method of gene tandem, and when the gene copy number reaches 3, the expression of AEP cyclase is the highest, reaching 0.89± 0.03mg / mL. The present invention adopts a new method to verify the cyclization ability of AEP cyclase. By constructing a tandem substrate, a protein that can be better observed in SDS-PAGE and has the possibility of cyclization is selected as the substrate , and construct a tandem substrate containing the substrate sequence of TEV protease, and identify whether it is cyclized according to the change in the size of the substrate after protease cleavage or the number of bands, the operation is simple, convenient, and practical.
Owner:湖北凯利特生物科技有限公司
Preparation method for beta-lactamase inhibitory polypeptide and expression vector used therein
InactiveCN102443593AReduce the difficulty of purificationReduce purification costsAntibacterial agentsFungiAntibody Affinity ChromatographyTEV protease
The invention discloses a preparation method for beta-lactamase inhibitory polypeptide and an expression vector, a fusion protein and the like used therein. The method comprises the following steps: 1) constructing a recombinant expression vector used for expressing His tag fusion protein which is a fusion protein of a TEV protease cleavage site His tag of beta-lactamase inhibitory polypeptide; 2) transforming host cells so as to obtain a transformant; 3) culturing the transformant and expressing the His tag fusion protein; 4) carrying out purification by using metal chelated affinity chromatography so as to obtain the His tag fusion protein; 5) carrying out restriction enzyme on the His tag fusion protein with TEV protease; 6) carrying out separation and purification by using gel column chromatography so as to obtain beta-lactamase inhibitory polypeptide. According to the invention, the effect of restriction enzyme of TEV protease is ideal, and TEV protease used in the method has a low price, thereby enabling both purification difficulty and cost of beta-lactamase inhibitory polypeptide to be substantially reduced.
Owner:SHANGHAI INST OF PHARMA IND
Expression vector, expression system and application of recombinant TEV protease
The invention relates to the field of biotechnology, and in particular to an expression vector, expression system and application of a recombinant TEV protease. The recombinant TEV protease expressionvector is obtained by inserting a His-TEVp recombinant protein into a pET-21a vector. The recombinant TEV protease expression system comprises introducing the recombinant TEV protease expression vector into E. coli to obtain a recombinant TEV protease expression strain. The His-TEVp recombinant protein is generated by inserting a four-point mutant TEVp gene into the pET-21a plasmid to obtain a recombinant plasmid pET-21a-TEVp. The TEV protease expression system is utilized to express the TEV protease, and then a large amount of high-purity TEV protease can be obtained by extraction and purification; the purity of the TEV protease can reach more than 90%; and the concentration of TEVp in the production bacterial liquid can reach 1.42 mg / ml, and high-purity TEV protease of more than 40 mg can be obtained in each liter of bacterial liquid. The expression vector and expression system of the recombinant TEV protease provided by the present invention lay a foundation for the popularizationand application of the TEV protease.
Owner:GUIZHOU INST OF TECH
A method for recombinantly expressing and producing human thymosin in yeast
ActiveCN102925470BEnsure product quality consistencyLow costHybrid peptidesVector-based foreign material introductionRestriction enzyme digestionNucleotide
The invention provides a method for massively producing human thymosin with low cost. The method is used for implementing restriction enzyme digestion to obtain a fusion gene sequence with a structure as follows: A-X-C, wherein A is a nucleotide sequence of site (1-150)-(1-372) amino acids at the N- end of coded mature human serum albumin; X is the nucleotide sequence of connecting peptide with enterokinase or tobacco etch virus (TEV) protease cutting site contained in a code; and C is a human thymosin gene. The method comprises steps as follows: connecting the fusion gene sequence to an expression carrier; transforming and introducing the expression carrier into saccharomycetes; carrying out induction expression to obtain soluble human serum albumin-thymosin fusion protein; then adding the TEV protease for cutting; and separating and purifying to obtain the recombined human thymosin. The human thymosin production method provided by the invention has the advantages that consistency of quality of products can be ensured, no limitation is generated from sources of raw materials, cost is low, an expression index is high, and mass production can be achieved and the like.
Owner:冯鹏波
A kind of recombinant type I humanized collagen polypeptide and its preparation method and application
ActiveCN113621052BIncrease productionPromote cell adhesionCosmetic preparationsBacteriaCell adhesionProteinogenic amino acid
The invention discloses a recombinant type I humanized collagen polypeptide and its preparation method and application. The recombinant type I humanized collagen polypeptide provided by the present invention comprises n repeats of the sequence shown in SEQ ID No.1, n is an integer greater than or equal to 1, wherein when n is an integer greater than or equal to 2, each repeat The sequences are directly connected; optionally, the N-terminal of the recombinant type I humanized collagen polypeptide contains an amino acid sequence that can be excised by TEV protease. The recombinant type I humanized collagen polypeptide provided by the present invention has the activity of promoting cell adhesion, the amino acid sequence of the recombinant protein is selected from the amino acid sequence of natural collagen, it will not produce an immune response when applied to the human body, and its preparation method is simple, Higher yields of collagen can be obtained at low cost.
Owner:SHANXI JINBO BIO PHARMA CO LTD
Syy capturer mutant, preparation method thereof and application of spy capturer mutant in fluorescent protein system
PendingCN114685679ASmall molecular weightReduced feasibilityFusions for enhanced expression stability/foldingFusion with protease siteLigationMutant
The invention discloses a spy capturer mutant, a preparation method thereof and application of the spy capturer mutant in a fluorescent protein system. Belongs to the technical field of biological coupling. According to the spyware catcher mutant, a flexible amino acid sequence is connected with the original N end and C end of a spyware SpyCatcher to obtain the spyware catcher mutant SpyCatcher-N, and a TEV protease recognizable sequence is inserted into the flexible amino acid sequence in the spyware catcher mutant SpyCatcher-N to obtain the spyware catcher mutant SpyCatcher-NTEV. The prepared spy capturer mutant has the advantages that on one hand, the high reactivity of a spy capturer is kept, and on the other hand, after a ligation reaction, Catcher protein can be digested by protease to reduce the molecular weight. Extra molecular weight is removed in a protease digestion mode.
Owner:江苏贝奥泰克生物科技有限公司 +1
Preparation method of high-purity cryptogein
The invention provides a preparation method of a high-purity cryptogein, which comprises the following steps: (1) sequentially connecting a coding gene of a target protein with TEV protease and T4 bacteriophage Hoc protein gene fragments to prepare a coding gene of the fusion recombinant protein; (2) transferring the coding gene of the fusion recombinant protein into a plasmid; (3) transferring the plasmid into an escherichia coli host; (4) infecting the escherichia coli host with Hoc-T4 bacteriophage deleted by Hoc protein, and culturing the host to express T4 bacteriophage displaying the fusion recombinant protein; (5) separating T4 bacteriophage displaying fusion recombinant protein; (6) separating the fusion recombinant protein from the T4 bacteriophage, and obtaining the target protein after TEV protease cleavage. The preparation method can simply and rapidly obtain high-purity cryptogein, and the obtained cryptogein solution can be directly used as an agricultural antibacterial bacteriostatic agent to achieve the effect of obviously promoting plant growth, thereby having good application prospect.
Owner:EZHOU INST OF IND TECH HUAZHONG UNIV OF SCI & TECH +1
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