235 results about "Mutant protein" patented technology

The invention provides an infectious bronchitis virus subunit vaccine as well as a preparation method and application thereof, and belongs to the technical field of genetic engineering vaccines. According to the chicken infectious bronchitis virus mutant protein disclosed by the invention, the amino acid sequence of the mutant protein is shown as SEQ ID NO.5. According to the avian infectious bronchitis virus subunit vaccine containing the recombinant avian infectious bronchitis virus mutant protein Kozak-IgKsp-IBV-M41-S-core-6His as an effective component, the glycosylation level of the vaccine is relatively high, the expression level is relatively stable, the virus return risk of a live vaccine does not exist, the safety is high, large-scale industrial preparation can be achieved, and the vaccine has good application prospects. The cost is low.

Disclosed are a new interleukin-2 (IL-2) mutant protein and the use thereof. Compared with wild-type IL-2, the IL-2 mutant protein has improved properties, such as an improved IL-2 receptor binding property and improved druggability. Also provided are a fusion protein, dimer and immunoconjugate comprising the IL-2 mutant protein, nucleic acids encoding the IL-2 mutant protein, the dimer and the immunoconjugate, and a vector and host cell comprising the nucleic acid. Further provided are methods for preparing the IL-2 mutant protein, the fusion protein, the dimer and the immunoconjugate, a pharmaceutical composition containing same, and the therapeutic use thereof.

The invention provides a method for improving the expression level of a meat toxin light chain. The method comprises the following steps: connecting a dissolution-promoting tag to an N end or a C end of meat toxin light chain protein to form soluble meat toxin light chain recombinant protein; the dissolution-promoting tag is a wild dissolution-promoting tag NT11 or a mutant protein mut7 of the wild dissolution-promoting tag NT11, enzyme digestion removal is not needed, the activity of a meat toxin light chain is hardly influenced and interfered, and the fusion protein can keep 93% of substrate enzyme digestion activity. The method further comprises overexpressing a molecular chaperone htpG in the genome of the host bacterium. According to the invention, a BoNT / A-LC high-efficiency expression system is constructed through multi-strategy optimization, so that the yield of BoNT / A-LC is increased by nearly 20% compared with that of wild type NT11-Lc, the yield reaches 647 mg / L through overexpression of a molecular chaperone htpG, and after combinatorial optimization, the yield is 8.5 times that of only expressed BoNT / A Lc, 1.79 times that of NT11-Lc and 1.43 times that of Mut7-Lc.

The application discloses application of SbWOX gene and SbBBM gene in corn breeding and a corn genetic transformation method, and relates to a bioengineering technology.The application provides a nucleic acid molecule with co-expression of SbWOX protein gene and SbBBM protein gene, and corresponding gene co-expression vectors and microbial transformants.The application also provides application of the co-expression of the SbWOX gene and the SbBBM gene in the corn genetic transformation method.The application obtains corresponding genes and mutant proteins based on the SbWOX gene and the SbBBM gene of sorghum, the two genes are co-expressed in the corn genetic transformation process, even without adding plant growth hormones in the antibiotic screening medium in the tissue culture process, the plant genetic transformation process can be normally completed, and the transformation efficiency and the regeneration efficiency of the corn can be remarkably improved, the transformation rate reaches 10.42%, and the differentiation rate reaches 32.89%.

The application discloses a five-heterocycle compound, a preparation method and application thereof. Specifically disclosed are a compound as shown in formula I, a pharmaceutically acceptable salt or solvate thereof, a composition containing the compound, and a pharmaceutical use of the compound for preparing a drug for treating a disease or disorder related to the mechanism of action of KRAS G12C mutant protein. The five-heterocycle compound in the application has the activity of inhibiting the proliferation of Ba / F3 KRAS-G12C cells, NCI-H358 cells and MIA PaCa-2 cells expressing KRAS G12C mutant protein, and has a good inhibitory effect on the growth of a subcutaneous transplanted tumor of a NCI-H358 nude mouse.

The invention discloses a lactic dehydrogenase LdLDH mutant and application thereof, a wild type LdLDH enzyme derived from Lactobacillus delbrueckii subsp. Bulgaricus is taken as a research object, the amino acid sequence of the wild type LdLDH enzyme is designed and modified to obtain a mutant protein, the mutant protein is named as LdLDH-M, the amino acid sequence of the mutant protein is shown as SEQ ID NO.2, the Tm value of the LdLDH-M is 77.1 DEG C, the Tm value of the LdLDH-M is 77.1 DEG C, and the Tm value of the LdLDH-M is 77.1 DEG C; compared with the wild type LdLDH enzyme, the Tm value of the LdLDH enzyme is increased by 27.4 DEG C, and the enzyme activity of the LdLDH enzyme is increased by 1.4 times. Therefore, compared with the wild type LdLDH enzyme, the LdLDH-M has remarkably improved thermal stability and enzymatic activity, and is more beneficial to widening the application range of the LdLDH-M in the fields of high-temperature biological research, industrial biological catalysis and the like.

A long-acting FGF21 derivative, specifically relating to an FGF21 derivative comprising an FGF21 mutant protein and a fatty acid chain. The FGF21 derivative, which is obtained by coupling the fatty acid chain to a cysteine residue at position 172 of an FGF21 protein by means of a linker, has relatively high FGF21 activity, and has a better weight loss effect in model mice. The FGF21 mutant protein not only retains the activity of mature FGF21, but also has significantly improved stability in serum.

A fusion protein comprising an anti-PD1 antibody moiety having a C-terminus, and an IL21 mutant protein conjugated to the anti-PD1 antibody moiety having a C-terminus wherein the IL21 mutant protein comprises at least one of two mutations relative to wild-type IL21: (1) one of a K72Y substitution, a K72M substitution and a K72Q substitution, and (2) a 75-80 deletion. In some examples, the anti-PD1 antibody may take the ETYY format and may comprise: a first heavy chain comprising a VH comprising the sequence of SEQ ID NO: 11; and a second heavy chain comprising a VH comprising a sequence selected from the sequences of SEQ ID NO: 8-10. Also provided are pharmaceutical compositions comprising the fusion protein, nucleic acids encoding the fusion protein, vectors comprising the nucleic acids, and methods of administering the fusion protein to treat diseases.

The present application relates to the application of Klenow mutant fragments in the synthesis of nucleic acids, and belongs to the technical field of biological modification and synthesis.The present application provides the application of Klenow mutant fragments with an amino acid sequence as shown in SEQ ID NO.8 or SEQ ID NO.14 in the synthesis of nucleic acids.The mutant protein F762A expressed by the Klenow mutant fragments of the present application can efficiently synthesize deoxyribonucleic acid, ribonucleic acid, deoxyribonucleic acid analogs and ribonucleic acid analogs with different lengths of DNA templates and DNA / RNA primers at 37 DEG C, and has a wide synthesis range and universality.

This invention pertains to fusion protein mutants comprising a Prime Editing enzyme having a first amino acid sequence and a second amino acid sequence, wherein the first amino acid sequence comprises a SpCas9 H840A nickase mutant protein of SEQ ID NO:152 and the second amino acid sequence comprises a Moloney Murine Leukemia Virus reverse transcriptase protein mutant (MMLV RTase mutant), wherein the fusion protein mutant displays at least the equivalent or greater activity of a reference Prime Editing enzyme in genome editing.

The application discloses a novel trimeric recombinant protein vaccine. Specifically disclosed are a mutant protein RBD8M with an amino acid sequence of SEQ ID No. 1 and a fusion protein of positions 26-277, positions 1-277 and SEQ ID No. 4. The application also discloses a vaccine containing the fusion protein, and further develops a trimeric recombinant protein vaccine containing the fusion protein and a double adjuvant. Experiments show that the vaccine prepared in the application can effectively induce cellular immunity and humoral immunity of the body in animals, a trace amount of the protein can stimulate a very high immune response of mice, has a good protection effect, can simultaneously produce high-titer neutralizing antibodies against various epidemic strains of the novel coronavirus, is a broad-spectrum effective new coronavirus vaccine, and has important significance for preventing novel coronavirus infection and a wide clinical application prospect.

The present invention relates to a novel interleukin-2 (IL-2) mutant protein. The present invention further provides a fusion protein and an immunoconjugate comprising the IL-2 mutant protein, a nucleic acid encoding the IL-2 mutant protein, and a vector and a host cell comprising the nucleic acid. The present invention further provides a method for preparing the IL-2 mutant protein, a pharmaceutical composition comprising the IL-2 mutant protein, and therapeutic use of the mutant protein.

Provided herein are novel IL-2 fusion proteins comprising an IL-2 moiety linked to an IL-2 masking moiety by a protease sensitive linker (PSL), the IL-2 masking moiety comprising an IL-13 mutein, an IL-13Ra2 binding mutein, an IL-13Ra2 antibody or antigen binding fragment thereof, an extracellular domain of CD122, an extracellular domain of CD132, or an extracellular domain of CD25. In some embodiments, the IL-2 fusion protein comprises an IL-13 mutein or an IL-13Ra2 antibody or an antigen binding fragment thereof, the IL-13 mutein or the IL-13Ra2 antibody or the antigen binding fragment thereof is capable of binding to IL-13R [alpha] 2, but not to IL-13R [alpha] 1, and the protease-sensitive linker can be cleaved by a protease in the tumor microenvironment. Such IL-2 fusion proteins are useful, for example, in the treatment of cancers that express IL-13R [alpha] 2.

This invention discloses a mutant of glycolaldehyde synthase and its application in improving the thermostability of glycolaldehyde synthase, belonging to the field of biocatalysis application technology. The glycolaldehyde synthase mutant disclosed in this invention, based on the amino acid sequence shown in SEQ ID NO.1, contains the following mutations: A381P / E509F, A381P / K290P, A381P, S61A, A381P / K290P / E509F, A381P / C49A, A381P / T100V, A381P / S61A, or A381P / K290P / S61A. This mutant protein can catalyze the preparation of hydroxyacetaldehyde (glycoaldehyde) from formaldehyde. The mutant protein of this invention also improves the catalytic efficiency of the formaldehyde-to-glycoaldehyde conversion. Due to the improved thermostability, it is beneficial for continuous catalysis in actual industrial reactions, thus the mutant shows good prospects for industrial applications.

The invention discloses a functionalized targeting fluorescent nanoprobe and application thereof in protein mutation analysis, and relates to the technical field of biology, the fluorescent nanoprobe comprises a targeting molecule-PEG hydrophilic bridging molecule and a matrix; the preparation method comprises the following steps: adding nano-seeds into fluorescent molecules to prepare fluorescent nano-particles, and stirring and mixing the fluorescent nano-particles with targeting molecules-PEG hydrophilic bridging molecules; the method is combined with deep learning to be applied to protein mutation analysis. The multifunctional integrated protein mutation receptor targeted fluorescent nanoprobe provided by the invention can accurately mark the position of a mutant protein at a single cell level, and can realize in-situ, real-time and dynamic living cell imaging of a tumor cell endogenous molecular mutant protein; and qualitative and quantitative analysis of tumor cell protein mutation can be realized through subcellular localization of the targeted fluorescent nanoprobe.

Mutant Cas9 proteins are described that have one or more mutations in a Keap1 degron sequence of the Cas9 protein. The one or more mutations can increase the half-life of the mutant proteins. Nucleic acid sequences and constructs comprising a sequence that encode a mutant Cas9 protein, as well as methods of enhancing CRISPR efficiency by administering one or more guide RNAs (gRNAs) to a cell comprising the mutant Cas9 proteins are also disclosed herein.

This disclosure provides an E. coli strain, which lacks thioredoxin reductase activity encoded by trxB and thioredoxin 1 activity encoded by trxA, and glutathione reductase activity encoded by gor. Said E. coli strain expresses a mutated AhpC protein having glutathione reductase activity and a cytosolic prokaryotic disulfide isomerase. The E. coli strain has an oxidative cytosol and can be used to efficiently produce proteins having disulfide bonds.

The invention discloses a method, device and equipment for predicting the property of a mutant protein, and the method comprises the steps: carrying out the fusion processing of the embedding characteristics and physicochemical characteristics of an original protein, the embedding characteristics and physicochemical characteristics of the mutant protein generated by the mutation of the original protein, and the interaction characteristics of residues of the mutant protein, obtaining initial node characteristics of the mutant protein; the interaction characteristic represents the interaction between every two residues in the mutant protein; the embedded feature represents an amino acid sequence of the protein, the physicochemical feature represents physicochemical properties of residues forming the protein, and a prediction model is used for processing initial node features and structural features of the original protein so as to predict multiple properties of the mutant protein; and predicting model loss of the model, wherein the model loss is determined according to a sample property predicted value representing multiple properties and a plurality of sample property measured values of the sample, wherein the sample property predicted value and the sample property measured values are obtained by the prediction model.

The invention discloses a carbohydrate molecule sequencing method based on glycosidase and nanopores and a system and application thereof. The invention also discloses a nanopore mutant protein and application thereof. The invention also discloses a glycosequencer containing the nanopore protein, a sequencing method, a system and application thereof. The invention also discloses a data processing method and a processing system for glycosidase hydrolysis assisted nanopore sugar sequencing, a computer related product and the like. According to the method, the detection limit can be remarkably reduced, the sensitivity can be improved, the accuracy of sugar sequencing can be improved, the analysis accuracy exceeds 98%, the sensitivity is improved by more than 50 times, the time is saved by more than 5 times, the analysis period is short, the sequencing process is simplified, and the cost is reduced. And a new technical approach is provided for rapid analysis of complex sugar sequence information.

An NCg12747 gene mutant and the use thereof in the preparation of L-lysine are provided. The NCg12747 gene mutant is a DNA molecule shown in SEQ ID NO: 3, which encodes the NCg12747 mutant protein shown in SEQ ID NO: 4. Mutating the NCg12747 gene into the NCg12747 gene mutant shown in SEQ ID NO: 3 or overexpressing NCg12747 gene mutant contributes to increase in the yield and the growth rate of L-lysine. However, when the gene is weakened or knocked out, accumulation of the L-lysine is not facilitated, and the growth rate of a strain can be reduced. The NCg12747 gene mutant and the NCg12747 mutant protein encoded thereby can be used for preparing L-lysine.

The application discloses a mutant Cas12i nuclease and application thereof, and belongs to the technical field of gene editing. The Cas mutant protein provided by the application is a mutant protein in which the 7th, 168th, 273rd, 332nd, 478th, 505th and 551st amino acids of SEQ ID NO:1 are all mutated into arginine, and other amino acid sequences remain unchanged. The application improves the editing efficiency by mutating multiple amino acids of the wild-type Cas12i protein, and can be used for multi-gene editing.

A treatment composition for inhibiting systemic sclerosis Vimentin mutant protein activity by using an STAT6 inhibitor, which inhibits the expression of an M2 macrophage and a profibrotic T cell which are immunocytes related to systemic sclerosis, and increases the expression of a Treg, and inhibits the expression of TGF-β, Col1a1, and α-SMA which are fibrosis factors related to systemic sclerosis. The presence of a pSTAT6 expression CD8 T cell in a fibrosis tissue has been identified, and that the expression of a pSTAT6 expression CD8 T positive cell is controlled via injection of the STAT6 inhibitor. In an animal model with increased Vimentin-specific disease symptom activity, the STAT6 inhibitor inhibits antigen-specific tissue fibrosis of systemic sclerosis with activated disease symptoms, and that the STAT6 inhibitor inhibits the expression of IL-17 cytokine expression CD8 positive TRM capable of inducing fibrosis and cell inflammation which are systemic sclerosis diseases.

The present application relates to compounds of formula (A) that inhibit the growth of cancers that express G12D mutant Ras proteins. The molecules of the specification are thus indicated for use in the treatment of cancer that express G12D mutant Ras protein.

The invention provides a compound as shown in a formula (I) and a racemate, a stereoisomer, a tautomer, a nitrogen oxide, a solvate, a polymorphic substance, a metabolite, an ester, a prodrug or a pharmaceutically acceptable salt thereof, and the compound has a good p53-Y220C mutant activation effect and can be used for treating tumor diseases containing p53-Y220C mutant protein. And preparation of medicaments for such conditions or diseases.

The invention relates to construction of a proline hydroxylase alpha subunit mutant from a human placenta and an expression vector of the proline hydroxylase alpha subunit mutant, and a mutant protein which is efficiently expressed in Ecoli BL21 (DE3) and has greatly improved hydroxylation efficiency and a construction body of the mutant protein are obtained by analyzing and screening a natural hydroxylase subunit.

Provided is a mutant protease having improved cleaning performance against protein stains in a high-concentration detergent liquid. A mutant protease consisting of the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence having an identity of at least 90% with SEQ ID NO: 1 and comprising at least one selected from the group consisting of the following amino acid residues (a) to (g): (a) Arg or Lys at a position corresponding to position 191 of SEQ ID NO: 1; (b) Arg or Lys at a position corresponding to position 17 of SEQ ID NO: 1; (c) Ala or Phe at a position corresponding to position 141 of SEQ ID NO: 1; (d) Arg or Lys at a position corresponding to position 243 of SEQ ID NO: 1; (e) Arg or Lys at a position corresponding to position 300 of SEQ ID NO: 1; (f) Arg or Lys at a position corresponding to position 302 of SEQ ID NO: 1; and (g) Arg or Lys at a position corresponding to position 311 of SEQ ID NO: 1.

The invention discloses a method for detecting the conformational change of an APO-Cascade protein. Through mutation of Cas8 protein amino acid N376 and Cas5 protein K169 in a Casade protein, the obtained Cas8 and Cas5 labeled by non-natural amino acids can maintain functions, and at the same time, the mutant type of the Cas5 / 8 protein can be subjected to fixed-point fluorescence labeling; according to the method, the Cas6 is used as a template, the N end of the Cas6 is further modified with biotin, and a formed Casade compound can be fixed on a biotin-PEG functionalized single-molecule slide, so that the conformation state of the Casade protein can be truly reflected by using single-molecule fluorescence resonance energy transfer, an effective means is provided for in-vitro research on the structure-function relationship of a system in CRISPR / Casade, and the method has a wide application prospect. And the method is of great significance in designing the Cascade protein with optimized properties and enabling the Cascade protein to play new functions.

The present invention relates to a mutated transcription factor EB (TFEB) protein that loses the native exon 3. This protein is referred to herein as "TFEB-ex3" or "ex3-TFEB". The invention also relates to polynucleotides encoding such muteins; a vector comprising the polynucleotide; and a biomolecular means for removing TFEB exon 3 in a patient in need thereof. The invention also relates to a pharmaceutical composition comprising the above, for use in the treatment and / or prevention of lysosomal storage diseases and conditions characterized by lysosomal dysfunction.

The invention discloses a tylosin sensing biosensor, a construction method thereof and MphR mutant protein based on the tylosin sensing biosensor, and belongs to the technical field of gene engineering. The technical problem to be solved by the invention is how to detect tylosin. In order to solve the technical problem, the invention provides the biosensor for sensing tylosin, the biosensor can express an MphR31D1 protein and / or a coding gene containing the MphR31D1 protein, and the MphR31D1 protein can be a protein with an amino acid sequence of SEQ ID NO: 1. The invention also provides a preparation method of the biosensor, and application of the MphR31D1 protein and related biological materials thereof in detection of tylosin. According to the invention, escherichia coli Top10 is taken as a chassis bacterium, a macrolide compound transcriptional regulation factor MphR is taken as a starting template, and the tylosin whole-cell biosensor is successfully constructed through protein directed evolution.

Provided are an RSV F protein mutant, and a preparation method therefor and a use thereof. Provided is a mutant protein, the mutant protein being in the form of a monomer or a trimer, wherein the trimer is formed by three monomeric mutant proteins; the monomeric mutant protein is a fusion protein comprising a first polypeptide element derived from an RSV F protein and a trimerization domain element; in the first polypeptide element, a Pep27 fragment of the RSV F protein is replaced with a linker sequence; and the monomeric and trimeric mutant proteins are both bound by an antibody that specifically recognizes RSV pre-F. The mutant protein can stably remain in a pre-fusion state, can induce the production of specific IgG antibodies and neutralizing antibodies, has excellent immunogenicity, and can be used for the development of RSV vaccines.