129results about "Polypeptide with MBP-tag" patented technology

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 discloses a high-activity lipase mutant and application thereof in functional grease synthesis, and belongs to the technical field of enzyme engineering and the technical field of enzyme immobilization. According to the invention, the lipase from Burkholderia B. cepacian OCRI-Lip100 screened in the early stage is truncated and mutated, and the soluble expression efficiency in Escherichia coli is improved by adding a solubilizing tag at the C end, co-expressing with folding enzyme and the like. According to the lipase mutant prepared by immobilization of the DMONs nano material, the half-life period of the obtained immobilized lipase at 60 DEG C is prolonged, the catalytic activity relative to free enzyme is improved, and the lipase mutant has good industrial application potential.

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 discloses a saccharomyces cerevisiae strain for de novo synthesis of fucoxanthine precursor neoxanthine and application of the saccharomyces cerevisiae strain, and belongs to the technical field of biology. According to the recombinant saccharomyces cerevisiae disclosed by the invention, the 3-hydroxy-3-methylglutaryl coenzyme A reductase, the isopentenyl pyrophosphate isomerase, the farnesyl pyrophosphate synthase and the endoplasmic reticulum size regulation factor are expressed in an enhanced manner; according to the method, geranyl geranyl diphosphate synthase, phytoene dehydrogenase, 15-cis-phytoene synthase, bifunctional lycopene cyclase / phytoene synthase, beta-carotene hydroxylase, zeaxanthin epoxidase truncated mutant and violaxanthin deep oxidase-like protein are subjected to heterologous expression, so that the zeaxanthin deep oxidase-like protein is obtained; the ROX1 and GAL80 genes in the saccharomyces cerevisiae are knocked out, and the expression of squalene synthase ERG9 is down-regulated. When the strain is subjected to shake flask fermentation, the contents of neoxanthine and violaxanthin of the strain respectively reach 1.78 mg / L and 26.77 mg / L, so that the strain has a wide application prospect.

The present invention relates to research tools for structural biology, particularly for facilitating the determination of three-dimensional structures of biological macromolecules. More specifically, the present invention helps to improve the overall feasibility of structure determination, resulting in higher resolution and better quality of three-dimensional structures of proteins through complex formation with novel fusion polypeptides.

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 application discloses a preparation method of a recombinant ternary fusion protein dressing with epidermal cell activation function. The dressing is a fusion protein designed based on the research on the functions and properties of COL3A1 and IFN-kappa proteins by international peers and the laboratory. The specific method is that the fusion gene fragment of the covalent connection of COL3A1 and IFN-kappa with GGSGG as a linker is obtained by using overlap extension PCR technology, and a 6×His label is added. The target gene is cloned into a prokaryotic expression vector pET30 (modified in the laboratory) to obtain the fusion protein COL3A1-IFN-kappa with an MBP label. After expansion culture, the MBP-IFN-kappa-COL3A1 with a purity of more than 90% can be obtained through Ni-NTA column affinity chromatography. We verified the resistance of the fusion protein to salmonella in THP-1 cells, and verified the effect of the fusion protein on promoting wound healing and resisting salmonella infection on mice. Finally, it is proved that the MBP-IFN-kappa-COL3A1 has good uniformity, is easy to purify, has high activity, has no side effects, and is very suitable for being used as an epidermal cell activation dressing.

The invention relates to fusion proteins comprising encapsulin and a second polypeptide comprising LanM and / or MBP, encapsulins, protein cages derived from the same, and compositions and methods of use thereof.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to plant chlorophyll hydrolase as well as a variant, a coding gene and application thereof. The sequence of the chlorophyll hydrolase CLD2 is as shown in SEQ ID NO. 1, and the sequence of the chlorophyll hydrolase CLD3 is as shown in SEQ ID NO. 2. Compared with the prior art, the research finds that the CLD1, CLD2 and CLD3 proteins can hydrolyze chlorophyll a, chlorophyll b and pheopephyll a, and compared with the chlorophyll a and chlorophyll b, the three enzymes have the highest hydrolytic activity on the pheopephyll a and the lowest hydrolytic activity on the chlorophyll b. The hydrolysis capacity of the CLD3 is the strongest, the hydrolysis capacity of the CLD2 is 60% of the CLD3, and the hydrolysis capacity of the CLD1 is the weakest of the CLD1 is 15% of the CLD3. The chlorophyll hydrolase (especially CLD3) provided by the invention can efficiently degrade chlorophyll and generate a large amount of hydrolysates to make up for the production requirements of the current industry and pharmaceutical industry and make up for the defects of the prior art, so that the chlorophyll hydrolase has wide application prospects.

The present invention relates to polypeptides for degrading polyesters. Disclosed herein are microbially-derived polypeptides and engineered variants thereof for degrading polyethylene terephthalate (PET) and other polyesters. Also provided are methods of producing the polypeptides and methods for polyester degradation using the polypeptides.

The invention provides a p53 fusion protein based on a targeted colorectal cancer marker CEA and application of the p53 fusion protein in preparation of medicines for inhibiting colorectal cancer, and relates to the field of biological medicines. The fusion protein comprises any one of the following components: p28-p53, MBP-TEV-p14ARF (1-63)-linker-p28, p28-p53-CEABP1, CEABP1-p28-p53, and CEABP2-p28-p53, and the fusion protein comprises any one component selected from the group consisting of the following components: a protein A, a protein B, a protein A, a protein B, a protein C and a protein B, according to the application, p53 and p14 ARF proteins for inhibiting cell proliferation in a human body, cell-penetrating peptide and designed protein CEABP1 or CEABP2 of a targeted binding colorectal cancer marker CEA are fused for the first time, and cell experiments and mouse experiments prove that the protein has a relatively high function of inhibiting growth of colorectal cancer cells, does not influence normal cell growth and has a wide application value.

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 present disclosure relates to the use of a maltose dependent degron to control stability of a protein of interest fused thereto at the post-translational level. The present disclosure also relates to the use of a maltose dependent degron in combination with a maltose-responsive promoter to control gene expression at the transcriptional level and to control protein stability at the post-translational level. The present disclosure also relates to the use of a stabilization construct that couples expression of a cell-growth-affecting protein with the production of non-catabolic compounds. The present disclosure further relates to the use of a synthetic maltose-responsive promoter. The present disclosure further provides compositions and methods for using a maltose dependent degron, a maltose-responsive promoter, and a stabilization construct, either alone or in various combinations, for the production of non-catabolic compounds in genetically modified host cells.

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 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 provides a preparation method and application of MBP-GL fusion recombinant protein. A C3 region of SPG and a B structural domain of protein L are connected in series, codon optimization is performed on a gene sequence, the front end of MBP is introduced into a BamHI restriction enzyme cutting site, a cysteine sequence, a termination codon TAA and an EcoRI restriction enzyme cutting site consistent with an expression vector are added to the tail end of the B region of the protein L, and the MBP-GL fusion recombinant protein is obtained. BamHI and EcoRI are subjected to double enzyme digestion and then are connected to a carrier subjected to the same double enzyme digestion, a recombinant protein MBP-GL gene expression carrier of a structural domain of strongest binding IgG of protein G and protein L connected with MBP is constructed, the carrier is subjected to transformation, induced expression and purification, and the recombinant protein MBP-GL gene expression carrier which not only has glucan ligand specific binding capacity, but also can be used for preparing the recombinant protein MBP-GL gene expression carrier. And the recombinant protein can be combined with all antibodies, subtypes and antibody fragments with high load.

The invention belongs to the technical field of polypeptide preparation and biological medicine, and particularly relates to a preparation method of Jingzhaotoxin polypeptide IX (JZTX-IX) and application of Jingzhaotoxin polypeptide IX in treatment of diabetes mellitus. A chilobrachys jingzhao toxin polypeptide IX coding gene, a 6 * His tag nucleotide sequence, a thrombin restriction enzyme cutting site nucleotide sequence and an MBP hydrotropy tag nucleotide sequence are connected to a prokaryotic expression vector for prokaryotic expression, and the expressed polypeptide is purified to obtain the chilobrachys jingzhao toxin polypeptide IX protein. The target polypeptide with high purity, complete disulfide bond folding and uniform conformation is obtained, so that the high purity and biological activity of the target polypeptide are ensured. On the basis, the treatment activity of the JZTX-IX on diabetes mellitus especially type II diabetes mellitus is found and verified, and the JZTX-IX can remarkably reduce fasting blood glucose of mice with the type II diabetes mellitus, improve glucose tolerance and insulin sensitivity, relieve polydipsia and polyuria symptoms and promote insulin secretion and islet cell proliferation.

The present invention relates to anti-human Dectin-1 antibody clone 2M24 (hIgG4) or 15E2, and multispecific binding molecules comprising same, as well as methods for producing and using same.

The invention relates to the technical field of biology, and particularly discloses a preparation method and application of recombinant A-type botulinum toxin. According to the method disclosed by the invention, after the recombinantly expressed protoxin is combined with cation exchange filler, enzyme digestion and activation are realized by utilizing single protease, and then purification is carried out. The preparation method disclosed by the invention has the advantages of good convenience and small batch-to-batch difference, and the prepared recombinant A-type botulinum toxin also has obvious advantages in protein effectiveness and biological activity.

The invention provides compositions and methods for nucleic acid synthesis, including ordered and continuous complementary DNA (cDNA) synthesis across non-continuous templates using a modified eukaryotic non-long terminal repeat reverse transcriptase (non-LTR RT) protein.

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.

Described herein are isolated monovalent antibodies or antibody fragments thereof that bind human properdin. Such antibodies are useful in methods of treatment for diseases mediated by alternative complement pathway dysregulation.

The present invention belongs to the technical field of phage display peptide libraries, and specifically relates to a diethylstilbestrol mimotope peptide, a preparation method thereof and an application. The amino acid sequence of the antigen mimotope peptide of diethylstilbestrol is YVYPQMP. When applied, the antigen mimotope can be used for immunological detection by biologically synthesizing the antigen mimotope peptide, or the phage displaying the diethylstilbestrol mimotope can be directly obtained through phage amplification for immunoassay, or the diethylstilbestrol mimotope can be separated from the phage to replace the diethylstilbestrol standard for immunological detection and analysis. The detection process is simple and convenient, the result is accurate, and the sensitivity is high. The present invention effectively improves the detection sensitivity of diethylstilbestrol in food, and at the same time reduces the harm to the human body and the ecological environment during the detection process of diethylstilbestrol, and has outstanding application value.

To provide an anti-dectin-1 antibody and a method for using the same. [Solution] The present invention relates to an anti-human dectin-1 antibody clone 2M24 (hIgG4) or 15E2, and a multispecificity binding molecule containing the same, as well as methods for producing and using the same. Disclosure relates to an antibody that binds to human dectin-1, a multispecificity (e.g., bispecificity) binding molecule, and related methods for using and producing the same. Disclosed herein is a targeted phagocytic method for removing disease-causing substances, including host cells / host cell products, microorganisms, or their products, by administering a multispecificity (e.g., bispecificity) binding molecule comprising a dectin-1 binding arm and a second arm that specifically binds to the substance.

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.

The present invention relates to a NLRP3 variant comprising at least a partial deletion of the PYD. The present invention further relates to a method of screening for a compound that inhibits the NLRP3 inflammasome. The present invention further provides a method comprising the step of administering a compound identified by such a screening method to a subject with a NLRP3-associated inflammatory disorder.

Provided are compositions comprising leptin and methods of using the same to support weight loss. Also provided are modified leptins, including recombinant leptins, and methods of making and using the same.