85results about "Polypeptide with MBP-tag" patented technology

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 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 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.

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 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.

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.

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.

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.

The present invention relates to methods of treating a disease, and methods for reduction of the formation of anti-drug antibodies (ADAs) in response to the administration of a therapeutic agent. The invention further relates to methods of treating a disease, particularly a B-cell proliferative disorder, and methods for reduction of adverse effects in response to the administration of a therapeutic agent, particularly a T-cell activating therapeutic agent.

The invention provides an NFT protein, a preparation method and application thereof, and a purification method of NusA or NusA fusion protein, and belongs to the field of recombinant protein expression and purification. The NFT protein provided by the invention has a core function of specifically binding protein purification tag NusA protein, and can be used as a key tool for targeted binding and efficient purification of NusA fusion protein. Compared with a traditional NusA fusion protein purification method which depends on additional addition of 6 * His and other purification labels, the NFT protein can be combined with the NusA protein in a high-affinity and specific manner, the problem of impurity residues of a purified product caused by non-specific combination can be effectively avoided, and the purification purity and recovery rate of the NusA fusion protein are remarkably improved. The NFT protein provided by the invention provides a brand new solution for efficient purification of NusA fusion protein, and has important application value in the fields of research and development of recombinant protein drugs, industrial enzyme production, life science research and the like.

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.

Disclosed is a method of producing a non-naturally occurring peptide for reducing nitrogen, the method comprising: a) identifying a first group of peptides each having a binding pocket similar to a naturally occurring iron-sulfur cluster cofactor binding pocket of a naturally occurring nitrogenase, wherein each of the first group of peptides is not a nitrogenase or a subunit or domain of a nitrogenase; b) selecting, from the first group of peptides, a second group of peptides each having at least one target amino acid in the binding pocket; c) selecting, from the second group of peptides, a third group of peptides each having at least one target property; and d) modifying the sequence and / or structure of each of the third group of peptides to improve binding of the iron-sulfur cluster cofactor in the binding pocket. Also disclosed is a non-naturally occurring peptide for reducing nitrogen, comprising: at least one modification to a naturally occurring peptide; and a binding pocket for binding an iron-sulfur cluster cofactor; wherein the naturally occurring peptide is not a nitrogenase or a subunit or domain of a nitrogenase; and wherein the at least one modification improves binding of the iron-sulfur cluster cofactor in the binding pocket.

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.

The present invention relates to an NLRP3 variant comprising an at least partial deletion of PYD. The invention also relates to a method for screening the compound for inhibiting the NLRP3 inflammasome. The invention further provides a method comprising the step of administering to a subject suffering from an NLRP3-related inflammatory condition a compound identified by such a screening method.

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 herein, among other things, is a conjugate comprising a polymerase and a nucleoside triphosphate, where the polymerase and the nucleoside triphosphate are covalently linked via a linker that comprises a cleavable linkage. A set of such conjugates, where the conjugates correspond to G, A, T (or U) and C is also provided. Methods for synthesizing a nucleic acid of a defined sequence are also provided. The conjugates can also be used for sequencing applications.

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.

The application discloses a high-activity lipase mutant and application thereof in synthesis of functional oil, and belongs to the technical field of enzyme engineering and the technical field of enzyme immobilization. B. cepacian The lipase from OCRI-Lip100 is truncated and mutated, and the soluble expression efficiency in E. coli is improved by adding a solubility tag at the C terminal and co-expressing with a foldase. The lipase mutant prepared by immobilization with DMONs nanomaterials has a prolonged half-life at 60 DEG C and improved catalytic activity relative to free enzyme, and has good industrial application potential.

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