215 results about "Wild type protein" patented technology

Provided is a compound useful as an active ingredient in a pharmaceutical composition for treating cancer.　The present inventors have examined a compound useful as an active ingredient in a pharmaceutical composition for cancer treatment, and have completed the present invention by finding that: a heterocyclic compound represented by formula (I) and formula (I-I) has excellent an action of inducing degradation of a mutant KRAS protein and a wild-type KRAS protein, and in particular, an action of inducing degradation of a KRAS protein having a G12C mutation, G12D mutation, G12V mutation, or G13D mutation; and said heterocyclic compound has mutant KRAS inhibition and the inhibitory activity of KRAS amplified by a wild-type KRAS gene, and in particular, inhibitory activity of KRAS having a G12C mutation, G12D mutation, G12V mutation, or G13D mutation. The heterocyclic compound according to the present invention or a salt thereof can be used as a therapeutic agent for cancer.

The present invention belongs to the technical field of bioengineering. Provided are an IscB mutein and the use thereof. Compared to the wild-type IscB protein, the IscB mutein comprises arginine at at least one of the following amino acid positions: position 84, position 96, position 102, position 111, position 159, position 368 and position 386; preferably, the IscB mutein further comprises arginine at position 401 and position 456. The relatively strong interaction between the IscB mutein and a double-stranded DNA target improves the editing activity for mammalian cell lines, which can greatly enhance the application potential of the IscB mutein as a new-generation underlying tool for gene editing, and promote the use of the IscB mutein in the aspects of precision medicine, animal model construction and crop breeding.

The present invention provides a phosphorylated Tau protein mutant and application thereof. The provided phosphorylated Tau protein mutant comprises at least one mutation substitution or deletion at E117, A145, G204, S305, A384, and L408 relative to the amino acid sequence SEQ ID NO.1 of the phosphorylated wild-type Tau protein, has improved thermal stability, and the thermal stability is significantly higher than that of the wild-type Tau protein during long-term storage. The phosphorylated Tau protein mutant can be used as a standard product and has greater utilization value.

The invention provides a system and a method for screening Cas protein mutants capable of identifying different PAM sequences, and belongs to the technical field of bioengineering. The system comprises a Cas protein mutant screening vector and an sgRNA expression vector, the Cas protein mutant screening vector comprises the following expression elements: a Cas protein coding gene, a plasmid replicon and a first resistance screening tag expression cassette; the sgRNA expression vector comprises the following expression elements: a plasmid replicon, a second resistance screening tag expression cassette, a third resistance screening tag expression cassette and an sgRNA expression cassette. According to the system and the method, aiming at the problem that the efficiency of identifying PAM sequences except NGG by wild type spCas9 protein is low, an active Cas9 mutant capable of identifying PAM sites except NGG is screened out, and the application field and the editing efficiency of a CRISPR gene editing system can be greatly expanded.

The invention provides an Akkermann Amuc1100 protein mutant as well as a preparation method and application thereof, wild Akkermann Amuc1100 protein is subjected to mutation modification through a molecular biological technical means, and the bioactivity of the Amuc1100 protein mutant subjected to molecular modification is obviously improved compared with that of the wild protein; compared with a wild type Amuc1100 protein, the wild type Amuc1100 protein has a better effect when being applied to the aspects of regulating intestinal immunity, relieving acute pancreatitis, promoting browning of white adipose cells, inhibiting obesity, relieving ulcerative colitis and the like, and a very good application prospect is provided for developing bioactive medicines with higher efficiency.

The invention relates to the field of plant molecular biology and forest genetic engineering, and particularly provides a PagRAP2.3 protein point mutant and application thereof in drought resistance of poplar. The mutant PagRAP2.3 MA is obtained by mutating methionine at the first site and cysteine at the second site of a wild type PagRAP2.3 protein into methionine and alanine. The invention further discloses a preparation method of the mutant PagRAP2.3. An expression vector containing the PagRAP2.3 MA gene is constructed, poplar 84K is transformed through an agrobacterium-mediated method, and a transgenic line with stable expression is obtained. Functional verification results show that overexpression of PagRAP2.3 MA can significantly increase the plant height of the poplar and promote plant growth, but the sensitivity to moisture is enhanced under drought stress. The mutant can be used for regulating and controlling the growth and development of forest trees and evaluating the drought resistance, and has a forestry breeding application prospect.

The invention discloses a lytic polysaccharide monooxygenase mutant as well as a preparation method and application thereof. In order to solve the problems of low expression level, poor stability and the like of the existing lysing polysaccharide monooxygenase in a microbial expression system, the lysing polysaccharide monooxygenase RsLPMO9 is mutated to obtain a multi-site mutant of which the amino acid sequence is shown as SEQ ID NO.2, and the expression quantity of the multi-site mutant is obviously higher than that of a wild type protein RsLPMO9. The analysis of the thermal stability shows that the thermal stability of the multi-site mutant is obviously improved compared with that of the wild protein RsLPMO9, and the lytic polysaccharide monooxygenase mutant provided by the invention has an application prospect in the aspects of degrading biomass or plant polysaccharide and the like.

The invention discloses a recombinant protein for detecting an anti-myelin sheath related glycoprotein MAG antibody and a detection reagent of the recombinant protein, and belongs to the technical field of neural immunodiagnostics, five amino acid sites in MAG protein are subjected to specific amino acid point mutation replacement to obtain an MAG protein mutation sequence, the MAG protein mutation sequence is fused with GlcATP enzyme and HNK-1ST enzyme full-length sequences to construct a lentiviral expression vector, and the recombinant protein and the detection reagent are used for detecting the anti-myelin sheath related glycoprotein MAG antibody. According to the present invention, three genes are constructed, and are stably transfected in a host cell so as to stably express the three genes in one cell at the same time, such that the constructed recombinant protein and the wild type MAG protein have similar anti-MAG IgM antibody binding domain, and on the premise of not using GlcATP enzyme and HNK-1ST enzyme small molecule agonists, the sensitivity and the specificity of the detection of the MAG autoimmune antibody by using the CBA method are improved; the method is suitable for preparing related detection reagents for in-vitro detection of the IgM type anti-MAG antibody.

The invention relates to the field of nucleic acid editing, in particular to the technical field of regularly clustered interval short palindromic repeat (CRISPR). In particular, the invention relates to a mutant of a Cas effector protein, a fusion protein comprising the mutant, and nucleic acid molecules encoding the same. The invention also relates to complexes and compositions for nucleic acid editing (e.g., gene or genome editing). The invention also relates to methods for nucleic acid editing (e.g., gene or genome editing) using the proteins or fusion proteins comprising the invention. Compared with a wild type Cas protein, the Cas protein mutant disclosed by the invention has better activity, such as higher cleavage activity, stronger target site recognition capability and higher target sequence editing activity.

The present invention relates to RSV F protein mutants and their applications, belonging to the field of vaccines. Based on the amino acid sequence of wild-type RSV F protein, the present invention has carried out modifications in multiple aspects and in various ways, including disulfide bond mutations, proline mutations, cavity filling mutations, electrostatic mutations, p27 sequence modifications, and / or trimerization domain modifications, and designed a variety of RSV F protein mutants. Then, through gene synthesis, expression, and ELISA detection, the present invention tested the pre-fusion conformation (pre-F) ratio and stability under high and low temperature conditions of the RSV F protein mutants, and screened out several RSV F protein mutants with a relatively high pre-fusion conformation ratio and stable structure. Finally, candidate antigen representatives were selected for immunogenicity detection, which proved that they have good immunogenicity and can be used for the development of RSV vaccines, the preparation of RSV-specific antibodies, or RSV detection reagents.

The invention discloses Vip3 protein with improved activity and application thereof. Vip3Aa mutant protein is obtained by single-point mutation of a residue Y19 in a structural domain I or a residue V383 in a structural domain III; the Vip3Ca mutant protein is obtained by carrying out single-point mutation on a residue K383 in the structural domain III. The insecticidal activity of the Vip3 mutant protein disclosed by the invention on various lepidoptera pests is obviously improved compared with that of a wild protein.

The present invention relates to a MeCP2 gene-specific trans-splicing ribozyme, and a use thereof. The trans-splicing ribozyme according to the present invention replaces the RNA of a mutant MeCP2 gene, which causes Rett syndrome, with a wild type MeCP2 RNA, so as to reduce the expression of the mutant MeCP2 protein and increase the expression of the wild type MeCP2 protein, and enables regulated expression so that wild type MeCP2 expression can be prevented from exceeding physiological levels, and thus can be effectively used for preventing or treating Rett syndrome.

The invention aims to provide a method and equipment for predicting protein complex affinity change caused by mutation, and the method comprises the following steps: utilizing a first sub-network in a twin network to predict protein complex affinity change caused by mutation on the basis of wild type protein complex information and mutation information; determining wild type multi-modal coding information comprising wild type sequence coding information and discretely coded wild type geometric feature coding information; determining affinity information of the wild type through a decoder; determining mutant affinity information through the corresponding second sub-network; and determining affinity change information. By using discrete coding and multi-modal information, the model is helped to cope with the condition that the local structure is not well optimized, so that the model can obtain a more accurate affinity change prediction result by using complex information with low precision. And protein surface information is introduced, so that the model is helped to better learn the influence of a cavity formed by a protein interaction interface on the affinity change, and the affinity change prediction accuracy is further improved.

The invention relates to the technical field of biology and biology, in particular to a screening method of thermostable mutation sites of a G protein coupled receptor and application of the thermostable mutation sites of the G protein coupled receptor, and the screening method comprises the following steps: S1, replacing a third intracellular ring segment in a wild type G protein coupled receptor with a third intracellular ring sequence of a kappa opioid receptor; marking a SmBiT peptide fragment label on the wild type G protein coupled receptor; s2, mutating a wild type G protein coupled receptor according to a mutation site in the G protein coupled receptor mutant to be detected; s3, incubating the G protein coupled receptor modified in the step S2 at different temperatures to obtain modified G protein coupled receptor samples treated at different temperatures; s4, carrying out mixed incubation on the modified G protein coupled receptor samples treated at different temperatures and a nano antibody-LgBiT compound, then adding luciferase to act as a substrate, and detecting; the method is high in screening efficiency, low in cost and short in time.

The invention relates to an RSV F protein soluble fragment, an autotrimer structural domain and application thereof, and belongs to the technical field of recombinant protein. Based on an amino acid sequence of a wild RSV F protein, a soluble and rigid sequence is screened out, a soluble fragment is constructed, disulfide bond mutation pair modification is performed on the basis of the soluble fragment to obtain a self-derived trimer structural domain sequence, and the self-derived trimer structural domain sequence is used for replacing a transmembrane region and an intracellular region of the RSV F protein. Therefore, trimerization can be realized, stable pre-fusion conformation can be maintained, and soluble expression in cells can be realized at the same time. The invention also provides an RSV F protein mutant containing an autotrimer structural domain sequence, the generation and the stability of the pre-F conformation trimer are proved, a representative is selected from the pre-F conformation trimer for animal experiments, and it is proved that the pre-F conformation trimer has good immunogenicity and can be used for developing RSV vaccines and preparing RSV specific antibodies or RSV detection reagents.

The invention relates to the field of nucleic acid editing, in particular to the technical field of regularly clustered interval short palindromic repeat (CRISPR). In particular, the invention relates to a mutant of a Cas effector protein, a fusion protein comprising the mutant, and nucleic acid molecules encoding the same. The invention also relates to complexes and compositions for nucleic acid editing (e.g., gene or genome editing). The invention also relates to methods for nucleic acid editing (e.g., gene or genome editing) using the proteins or fusion proteins comprising the invention. Compared with a wild type Cas protein, the Cas protein mutant disclosed by the invention has better activity, such as higher cleavage activity, stronger target site recognition capability and higher target sequence editing activity.

The invention discloses an LbCas12a protein mutant as well as a preparation method and application thereof, the LbCas12a protein mutant is obtained by performing K390A or K945A mutation on a wild type LbCas12a protein, and the amino acid sequence of the wild type LbCas12a protein is as shown in SEQ ID NO. 1. According to the present invention, the LbCas12a protein is subjected to directional modification, the alanine mutation occurs at the K390 / K945 site, the LbCas12a-K390A protein mutant and the LbCas12a-K945A protein mutant are prepared, the protein mutants obtained based on the method provide a series of significant advantages in function, and the solid foundation is laid for the application of the protein mutants in multiple fields. Through Michaelis-Menten kinetic analysis, it is observed that when the LbCas12a-K390A / K945A protein mutant prepared through the method and the wild type LbCas12a protein target the same dsDNA target, the catalytic efficiency of the LbCas12a-K390A / K945A protein mutant is 42.1 times that of the wild type LbCas12a, and the catalytic efficiency of the LbCas12a-K390A / K945A protein mutant is 707.9 times that of the wild type LbCas12a, and the catalytic efficiency of the LbCas12a-K390A / K945A protein mutant is 707.9 times that of the wild type LbCas12a. The results show that K390 and K945 are mutated into alanine, so that the affinity of the LbCas12a protein to a substrate can be increased to a certain extent, and the LbCas12a protein has higher trans-cleavage activity.

The invention belongs to the technical field of characterization of target analytes, and provides application of PANX2 protein and a mutant thereof as biopore protein. The bioporous protein is used for detecting one or more of metal ions, inorganic salts, amino acids, nucleotides, small molecule drugs, small molecule diagnostic agents, adenosine triphosphate and monosaccharides. After one or more amino acids of the wild type PANX2 protein are mutated into common amino acids except original amino acids, compared with the wild type PANX2 protein, the PANX2 protein mutant can be normally expressed and is stable in property, a current signal is obviously improved during detection, the up-down amplitude of the current signal is longer, and burrs are fewer.

The present invention discloses mutants of wild-type RSV F protein. The binding experiment of pre-fusion conformation trimer-specific monoclonal antibody AM14 shows that the mutant provided in the embodiment of the present disclosure can have strong binding activity with AM14 monoclonal antibody, while the wild-type RSV F protein before artificial mutation has no binding activity with AM14 monoclonal antibody. The results of accelerated stability test show that the mutant provided in the embodiment of the present disclosure is placed in a 37°C environment for 4 weeks, and the binding activity of AM14 monoclonal antibody has no obvious change. The results of animal immunization experiments show that compared with the wild-type RSV F protein before artificial mutation, the mutant provided in the embodiment of the present disclosure can induce the production of higher titers of neutralizing antibodies.

The application discloses a LCC with enhanced hydrophobicity ICCG The application discloses a mutant protein and application thereof ICCG The mutant protein is obtained by mutating an amino acid residue at the 266th amino acid residue in a protein amino acid sequence into phenylalanine, and is a mutant N266F. Compared with a wild-type protein, the mutant protein has significantly enhanced hydrophobicity, and is verified by a PET depolymerization experiment. The mutant protein has significantly improved PET depolymerization efficiency compared with the wild-type protein under the condition of low enzyme load, is economically feasible, and has product selectivity.

This invention discloses an ANXA1 recombinant antigen, its preparation method, and its applications, belonging to the field of biodetection technology. The ANXA1 recombinant antigen of this invention is based on the wild-type ANXA1 protein shown in SEQ ID NO.1, obtained by deleting its first epitope-masking region, thus making the epitope more easily exposed. Verification has shown that, compared to the full-length ANXA1 antigen, the ANXA1 recombinant antigen of this invention has improved sensitivity in early lung cancer detection, and its detection performance is significantly superior to commercially available ANXA1 recombinant antigens. Therefore, the ANXA1 recombinant antigen of this invention provides a new approach for the early auxiliary diagnosis of lung cancer.

This invention discloses a PtCas12f variant protein, the amino acid sequence of which is shown in SEQ ID NO:1; a gene editing system containing the PtCas12f variant protein; a further modified denPtCas12f variant protein; and a gene expression regulation system containing the denPtCas12f variant protein. By modifying the wild-type PtCas12f protein to obtain the PtCas12f variant protein with the amino acid sequence shown in SEQ ID NO:1, the editing efficiency can be greatly improved, while the further modified denPtCas12f variant protein can greatly improve the gene expression regulation efficiency. Therefore, a novel and efficient gene editing tool is provided for gene editing and regulation, which can be widely used in the field of gene research.

The present application relates to the technical field of genetic engineering, and particularly relates to a low immunogenic LwCas13a protein mutant and application thereof.The LwCas13a protein mutant provided by the present application has one or more mutations of Q436C, K560D and K76C on the wild-type LwCas13a protein.The LwCas13a protein mutant provided by the present application significantly reduces the immunogenicity of the LwCas13a protein in a bovine body, thereby reducing the recognition and reaction of the immune system.The LwCas13a protein mutant provided by the present application not only does not affect the activity of the LwCas13a nuclease, but also maintains the effective expression and function of the LwCas13a nuclease in bovine cells.The gene editing efficiency of the LwCas13a protein mutant provided by the present application is equivalent to that of the wild type, and the interference efficiency is not significantly affected.

The invention relates to the technical field of gene engineering, in particular to a low-immunogenicity LwCas13a protein mutant and application thereof. According to the LwCas13a protein mutant disclosed by the invention, one or more mutations of Q436C, K560D and K76C are generated on a wild type LwCas13a protein. According to the LwCas13a protein mutant disclosed by the invention, the immunogenicity of the LwCas13a protein in a cattle body is remarkably reduced, so that the recognition and reaction of an immune system are reduced; the LwCas13a protein mutant disclosed by the invention not only does not influence the activity of LwCas13a nuclease, but also can maintain the effective expression and function of the LwCas13a nuclease in bovine cells; the gene editing efficiency of the LwCas13a protein mutant provided by the invention is equivalent to that of a wild type, and the interference efficiency is not obviously influenced.

The application belongs to the technical field of gene editing, and particularly relates to an optimized MmeFz2 protein and gene editing application thereof. The optimized MmeFz2 protein is obtained by mutating a wild-type MmeFz2 protein shown in SEQ ID NO. 1 as follows: mutating C at the 69th position of the wild-type MmeFz2 protein into K, mutating E at the 305th position into N and mutating E at the 326th position into Q; or mutating E at the 178th position of the wild-type MmeFz2 protein into H, mutating E at the 305th position into S and mutating E at the 418th position into R. The C-terminal of the optimized MmeFz2 protein is further fused with an HMG-D protein. The application further provides an optimized MmeFz2-omega RNA system, and the system has strong gene editing efficiency. The application has important significance and value for promoting the wide application of gene editing technology in functional mechanism research and clinical treatment.

The invention provides a pAPN protein mutant, a substance for mutating pAPN protein and application of the pAPN protein mutant, which are characterized in that on the basis of wild pAPN protein, amino acid of the pAPN protein is mutated into 220 D into G, or / and 918 F into P, or / and 570 V into I, or / and 595 H into Y, and other amino acid residues are not changed, so that TGEV SD / L strain infection can be effectively inhibited; a target spot is provided for obtaining a pig herd which can effectively inhibit TGEV infection and does not affect the growth performance of piglets and the physiological activity function of TGEV.

The invention provides a TRPM8 protein mutant and application thereof, relative to an amino acid sequence of a wild type TRPM8 protein, the TRPM8 protein mutant comprises mutation at one or more positions selected from the 825th site, the 864th site, the 891th site, the 897th site, the 915th site, the 917th site, the 925th site, the 927th site, the 928th site, the 932th site, the 945th site, the 946th site and the 948th site. The invention also provides a separated nucleic acid which is used for coding the TRPM8 protein mutant. The invention also provides a vector containing the isolated nucleic acid and a genetically recombinant porcine fetal fibroblast. According to the invention, a key site which can effectively reduce the cold activation level of the pig TRPM8 without influencing other activation functions is screened, and a new material is provided for animal breeding improvement.

The invention provides a protein modification method and device, electronic equipment and a storage medium. The method comprises the following steps: acquiring protein biological information of wild type protein; respectively inputting the protein biological information into a protein prediction model to obtain transformation probability distribution of initial mutation sites output by the protein prediction model; carrying out sampling search based on the transformation probability distribution to obtain transformed protein; the protein prediction model comprises a protein inverse folding model and a protein language model. The method provided by the invention comprises the following steps: respectively inputting protein biological information into a protein inverse folding model and a protein language model to respectively obtain transformation probability distribution of initial mutation sites output by the models; sampling search is performed based on the transformation probability distribution of the initial mutation sites to obtain the transformed protein of the wild type protein, so that the activity and stability of the transformed protein are greatly improved, the usability of the transformed protein is ensured, and the transformation efficiency of the protein is further improved.

The present invention provides TtdAgo mutant proteins with room-temperature target nucleic acid cleavage activity and their applications, belonging to the field of programmable nuclease technology. Relative to the wild-type TtdAgo protein, the TtdAgo mutant proteins have mutations at amino acids 527 and / or 561 and / or 593 and / or 599. Compared to the wild-type TtdAgo protein, the TtdAgo mutant proteins provided by the present invention not only have significantly improved activity at room temperature but are also capable of cleaving target RNA, effectively expanding the scope of application of the pAgo protein.

The invention provides a UvsX protein mutant, and compared with a wild type UvsX protein, the UvsX protein mutant has mutation at one or more of the 35th amino acid, the 39th amino acid, the 52th amino acid, the 56th amino acid, the 166th amino acid, the 169th amino acid, the 197th amino acid, the 198th amino acid, the 200th amino acid, the 211th amino acid, the 212th amino acid, the 214th amino acid and the 318th amino acid corresponding to SEQ ID NO: 4. Compared with a wild type UvsX protein, the UvsX protein mutant disclosed by the invention is high in yield and improved in activity.