172 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 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 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 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 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 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 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.

This invention belongs to the field of bioengineering and virus-like particle (VLP) construction technology, specifically relating to an amino acid sequence with steric hindrance effect and its preparation method, the VP2 protein of recombinant porcine parvovirus and its nucleotide sequence, and the virus-like particles assembled from it. An amino acid sequence with steric hindrance effect, SEQ ID NO: 1, is provided. This invention provides an amino acid sequence with steric hindrance effect, which is fused to the N-terminus of the wild-type VP2 protein to obtain a modified VP2 protein. During the critical phase of the assembly of the modified VP2 protein into VLPs, SEQ ID NO: 1 at the N-terminus, due to its steric hindrance effect, pre-occupies and physically seals the 5-fold pores of the capsid, ensuring that the host background nucleic acid cannot enter the particle interior. Through protein structure modification, nucleic acid entry into the particle interior is physically blocked from the source, resulting in a pure antigen with extremely high hollowing rate.

This invention belongs to the field of target characterization technology, and provides an application of PANX2 protein and its mutants as bioporins. The bioporins can detect one or more of the following: metal ions, inorganic salts, amino acids, nucleotides, small molecule drugs, small molecule diagnostic reagents, adenosine triphosphate (ATP), and monosaccharides. In this invention, one or more amino acids of the wild-type PANX2 protein are mutated to common amino acids other than the original amino acids. Compared to the wild-type PANX2 protein, the PANX2 protein mutants can be expressed normally, are stable, and show a significantly improved current signal during detection, with a longer amplitude and fewer spikes.

The invention relates to a method for improving enzyme activity by designing an oligomer through domain exchange and a mutant thereof. The method comprises the following steps: S1, replacing a hinge in a hinge database with software into a loop exposed on the surface of a wild type protein or truncating a loop region with different lengths to form a first batch of mutants; the hinge database comprises the following PDB ID structural domain sequences: 1K87, 4GW9, 1MPX, 1CQZ, 1BL9, 1NNO, 1DQU, 7CAT, 1M7S, 1CTS, 1KAH, 1K75, 3IVS, 1QE0, 2GYS, 2A5H, 2NZ2, 3O8J, 2BI4, 2HZK, 3MD0, 1XUU, 2WWW, 3MQ6, 1B4K, 2PAH, 1GZ6, 1U0T, 4NDJ, 1CR0, 1PYM, 3L3P, 1R8J, 1T98, 1IA9 and the like. The invention provides a simple and efficient oligomer design strategy for the field of protein engineering, not only lays a theoretical foundation for developing an enzyme preparation with higher catalytic activity and stability, but also provides an effective, universal and universal practical method for developing the enzyme preparation with higher catalytic activity and stability in the field.

The present invention relates to the field of biomedicine technology, and in particular to an α-myosin mutant and its application. Lysine (K) at position 1897 in the wild-type Myh6 protein is mutated to arginine (R). The nucleotide sequence corresponding to the wild-type Myh6 protein is shown in the bases in SEQ ID NO.1. Application of α-myosin mutants as Ang II-induced heart failure pathology models. The present invention obtains α-myosin mutants through the gene editing method of the CRISPR / Cas9 system, confirms that the 1897 site in α-myosin participates in and regulates the degree of interaction between α-MHC and Titin, thereby reflecting the degree of heart failure. The obtained α-myosin mutant is used as a pathology model leading to heart failure, which can lay the foundation for the study of organic heart failure, especially heart failure caused by abnormal myofilament structure.

The invention provides a detection reagent and a kit for detecting an anti-SRP antibody. The amino acid sequences of the recombinant SRP54 protein are as shown in SEQ ID NO: 1-3. According to the present invention, the wild type SRP54 protein is subjected to gene engineering modification, the stability of the modified recombinant protein (mutant type) is improved, the sensitivity and the specificity of the detection kit prepared by using the mutant type recombinant SRP54 protein for detecting the patient serum IgG are the same as the sensitivity and the specificity of the detection kit prepared by using the wild type SRP54 protein, and the thermal stability is improved; the attenuation of a detection signal value is slowed down, and the service life of the kit is prolonged.

The present invention provides a porcine epidemic diarrhea virus S protein mutant, a nucleic acid molecule, a subunit vaccine and applications thereof; compared with the wild-type porcine epidemic diarrhea virus S protein, the porcine epidemic diarrhea virus S protein mutant has the 1076th amino acid, the 1077th amino acid, the 1016th amino acid and the 1017th amino acid all mutated into proline. The present invention constructs a stable natural trimer that maintains the prefusion conformation of the S protein, obtains a safe and highly effective PEDV subunit vaccine, and can improve the use effect and immunogenicity of the porcine epidemic diarrhea vaccine; moreover, compared with the wild-type S protein, the porcine epidemic diarrhea virus S protein mutant in the present invention has a higher expression level.

This application discloses a composition, a method for preparing the composition, and its application, comprising: an mRNA molecule containing a first sequence; and an LNP nanoparticle delivery carrier; wherein the protein encoded by the first sequence has BP26 protein immunogenicity, the first sequence encodes a first amino acid sequence, the first amino acid sequence includes multiple sub-sequences in tandem, the multiple sub-sequences being selected from the amino acid sequences corresponding to T cell and B cell recognition epitopes in wild-type BP26 protein, and the composition has high immunogenicity and high targeting against Brucella.

The invention belongs to the technical field of medicines, and particularly relates to a milk acylation modified Myh6 protein and application thereof. The antigen polypeptide for identifying the heart failure marker has an amino acid sequence as shown in SEQ ID NO: 1, lysine located at the 9th site is subjected to milk acylation modification, and the heart failure marker is milk acylation modified Myh6 protein; the milk acylation modified Myh6 protein is a lysine milk acylation modified protein (Myh6-K1897) which is located at the 1897 site of a wild type Myh6 protein. The Myh6-K1897 can be used as a marker for identifying heart failure as a target spot, is a set of novel and perfect protein milk acylation molecular regulation and control system, can judge the interaction between Myh6 and Tin, and is used for treating the core and key of heart failure. The research provides a new and powerful treatment strategy for the treatment of human heart failure diseases.

The present invention discloses a recombinant protein for detecting anti-myelin associated glycoprotein (MAG) antibody and a detection reagent thereof, belonging to the technical field of neuro-immunodiagnosis. By performing specific amino acid point mutation substitutions at five amino acid positions in the MAG protein, a MAG protein mutant sequence is obtained, and then it is fused with the full-length sequences of GlcATPase and HNK-1ST enzyme to construct a lentiviral expression vector, which is stably transfected into host cells, enabling the simultaneous stable expression of the three genes in one cell. The constructed recombinant protein has a similar anti-MAG IgM antibody binding domain to the wild-type MAG protein, and without using small molecule agonists of GlcATPase and HNK-1ST enzyme, it improves the sensitivity and specificity of detecting MAG autoantibodies by the CBA method, and is suitable for preparing related detection reagents for in vitro detection of IgM-type anti-MAG antibodies.

The present invention relates to an ACCS gene, a protein and the use thereof. An involved ACCS protein mutant is formed by means of mutating a proline residue at position 134 and a tryptophan residue at position 136 of an animal-derived wild-type ACCS protein into a glutamine residue and a tyrosine residue, respectively. The involved ACCS protein mutant can be used in the intervention and treatment of a variety of diseases comprising brain diseases, and provides a new regimen and approach for the intervention and treatment of brain diseases.

The invention 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 performing any one of the following mutations on a wild type MmeFz2 protein with an amino acid sequence as shown in SEQ ID NO.1: 1) mutating C at the 69th site of the wild type MmeFz2 protein into K, mutating E at the 305th site into N, and mutating E at the 326th site into Q; and 2) mutating the E at the 178th site of the wild type MmeFz2 protein into H, mutating the E at the 305th site into S and mutating the E at the 418th site into R. The C end of the optimized MmeFz2 protein is also fused with an HMG-D protein. The invention further provides an optimized MmeFz2-omega RNA (Ribonucleic Acid) system, and the system has high gene editing efficiency. The method has important significance and value for promoting wide application of a gene editing technology in the aspects of functional mechanism research and clinical treatment.

This invention discloses a kit for detecting porcine reproductive and respiratory syndrome virus (PRRSV) antibodies and its applications, belonging to the field of animal disease detection technology. The kit uses a mutant nucleocapsid protein (N protein) of the PRRSV-2 class NADC30 strain CHsx1401 as the coating antigen; the N protein mutant has mutations at amino acid positions 23, 75, and / or 90 compared to the wild-type N protein. High purity is achieved through optimized prokaryotic expression systems and purification processes. Under optimal detection conditions, the kit exhibits strong specificity, high sensitivity, and good reproducibility, with significantly superior detection performance compared to existing commercial kits. It can be used for PRRSV-2 antibody detection, epidemiological surveys, and disease control.

Inventors followed an untargeted approach by screening a single chain variable fragment (scFv) library via phage display against recombinant human full-length wild-type (wt)TDP-43. They identified four wtTDP-43-specific scFv, two of which were retained following cellular expression and colocalization with TDP-43 in vitro. In silico binding site prediction on TDP-43 suggested the pathologically-relevant C-terminal and RRM1 domains as potential targets. One scFv diminished the amount of the insoluble 35-kDa C-terminal fragment of TDP-43 when the wildtype protein was overexpressed. Another scFv inhibited NF-κB activation associated with TDP-43 overexpression. Both scFv seemed to reverse some metabolic alterations caused by TDP-43 overexpression. Their findings offer two scFv molecules that bind wtTDP-43, alter its aggregation, and modify cellular pathways associated with TDP-43 proteinopathies. Accordingly, the present invention relates to scFv intrabodies targeting TDP-43 and their uses in diagnosis and treatment methods.