114 results about "Wild type protein" patented technology

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 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.

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 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.

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.

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.

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 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.

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.

The invention provides a phage E protein mutant and application thereof. Compared with the phage E protein, the phage E protein mutant has the advantage that the 33rd lysine is mutated into arginine. According to the invention, the cracking capability of the K33R variant under the background of the vibrio natriticus MraY is screened and proved to be obviously superior to that of a wild type E protein, so that the blank of lack of a standard cracking element for the vibrio natriticus in the prior art is filled; and an effect module which can be directly applied is provided for realizing stable oscillation of an ePop self-splitting line in the vibrio natriticus or engineering bacteria carrying the vibrio natriticus MraY.

A respiratory syncytial virus (RSV) vaccine is provided, the nucleic acid comprising a polynucleotide encoding a mutant of an RSV F protein, the mutant to comprising one or more of the following mutations relative to a wild-type RSV F protein: a disulfide bond mutation, a cavity filling mutation, and an electrostatic mutation.

This invention belongs to the field of bioengineering technology, specifically relating to a BmoR protein mutant capable of sensitively detecting n-butanol or isobutanol, and its application in n-butanol or isobutanol detection or biosensors. The BmoR protein mutant is obtained by mutating the wild-type BmoR protein shown in SEQ ID NO.1 to the E54G mutation. The E54G mutant, possessing the amino acid sequence shown in SEQ ID NO.3, is more sensitive to n-butanol (2.64 × 10⁻⁶). ‑6 mM) and isobutanol (2.16×10 ‑6 The method uses mM) detection to solve the problem of low sensitivity of wild-type BmoR protein, which cannot screen for extremely low concentrations of n-butanol or isobutanol. It can be used for screening and application of strains that produce low levels of higher alcohols.

The present application relates to the field of genetic engineering and biological enzyme depolymerization of PET plastics, and particularly relates to a hydrophilic PET depolymerase mutant and application thereof. The present application obtains a hydrophilic PET depolymerase mutant V68R, or A137R or I243R by mutating the 68th, or 137th, or 243th amino acid residue of the PETase protein amino acid sequence into R. Compared with the wild-type PETase protein, under the premise of ensuring the depolymerization efficiency of PET / BHET and effectively depolymerizing PET / BHET, the expression level of soluble protein is improved, and the formation of inclusion bodies is reduced in the expression system of Escherichia coli, Bacillus subtilis or Pichia pastoris, which has economic feasibility, PETase high-yield universality, and wide application prospect.

This invention belongs to the field of biotechnology, specifically disclosing novel mutations of ANKRD11 and their applications. The ANKRD11 gene mutation can be any of the following: a nucleic acid having a target fragment, wherein the target fragment has a G repeat at position 4708 compared to the wild-type ANKRD11 gene with sequence SEQ ID NO.1; or a polypeptide having the p.E1570Gfs*71 mutation compared to the wild-type protein encoded by the ANKRD11 gene with sequence SEQ ID NO.2. The invention also relates to the application of reagents for detecting the aforementioned ANKRD11 gene mutation in screening individuals at risk for KBG syndrome. This disclosure broadens the pathogenic gene spectrum of KBG syndrome, enhances the understanding of the disease, provides experience for clinical screening and diagnosis of the disease, and also provides a basis for prenatal diagnosis.

The invention belongs to the technical field of tobacco gene engineering, and particularly relates to a CCD4 protein mutant G312S and application thereof. Compared with a wild type CCD4 protein, the mutant has the following mutation: p. G312S, and the NCBI (National Center of Biotechnology Information) accession number of the wild type CCD4 protein is AKO22632.1. According to the invention, the substrate beta-carotene is docked to a CCD4 enzyme activity pocket, a saturated mutant library is constructed, screening is carried out through a bacterial color complementation experiment, and after specific site amino acid mutation, degradation of the substrate beta-carotene is accelerated, so that a certain technical foundation is laid for further cultivation of new varieties of tobacco crops.

The invention belongs to the technical field of gene editing, and particularly relates to an IscB / omega RNA gene editing mutant, an editing system and application. According to the invention, a wild-type IscB protein and a wild-type omega RNA skeleton are respectively truncated to obtain an IscB mutant and an omega RNA skeleton mutant. Compared with a wild type IscB protein, the IscB mutant provided by the invention is smaller in size, higher in editing efficiency and lower in off-target efficiency; compared with a wild-type omega RNA skeleton, the omega RNA skeleton mutant provided by the invention is more compact and keeps high editing ability at the same time. An IscB / omega RNA gene editing system (IscB # / omega RNA #) established based on the omega RNA skeleton mutant and the IscB mutant has high editing efficiency and low off-target efficiency, and realizes verification in different cell lines in vitro.

This invention belongs to the field of bioengineering technology, specifically relating to a method for... R,R BmoR protein mutants with a wider detection range (0-200 mM) for 2,3-butanediol, and their applications in... R,R Applications in the detection or biosensor of 2,3-butanediol. The BmoR protein mutant was obtained by mutating the wild-type BmoR protein shown in SEQ ID NO.1 with the N89D mutation, and has the amino acid sequence shown in SEQ ID NO.3. The N89D mutant... R,R -2,3-Butanediol has a wider detection range (0-200 mM), solving the problem of the inability to screen wild-type BmoR proteins. R,R The problem of high-yield strains of 2,3-butanediol can be addressed by screening and applying strains with even higher yields.

According to the phosphorylated protein-polypeptide conjugate and the preparation method thereof, polypeptide in the phosphorylated protein-polypeptide conjugate has threonine (Thr) phosphorylation modification, phosphorylation coupling sites can be accurately positioned, the phosphorylated protein-polypeptide conjugate has high thermal stability, and the thermal stability of the phosphorylated protein-polypeptide conjugate is remarkably higher than that of wild type Tau protein in long-term storage. The method can be used as a quality control product and has a greater utilization value.

The invention provides an IL-2 variant, an IL-2 variant and PD-1 antibody fusion protein and application of the IL-2 variant and PD-1 antibody fusion protein, specifically, the fusion protein comprises a first binding region, the first binding region is provided with the IL-2 variant, and compared with wild type IL-2 protein, the IL-2 variant has at least one of the following site mutations: the 3rd site amino acid, the 39th site amino acid, the 88th site amino acid and the 125th site amino acid. The IL-2 variant is weakly combined with IL2R alpha and extremely weakly combined with IL2R beta gamma, and compared with IL-2 wild type mutants and other types of mutants, the variant has higher activity and lower immunotoxicity and is not prone to causing immune storm, and therefore the IL-2 variant has the advantages that the IL-2 variant can be used as an immunopotentiator, and the IL-2 variant can be used for immunopotentiator immunopotentiator for immunopotentiator immunopotentiator immunopotentiator for immunopotentiator immunopotentiator immunopotentiator. Compared with fusion proteins prepared from IL-2 wild type and other types of mutants, the fusion protein prepared from the IL-2 variant has a stronger anti-tumor immune function, is lower in immunotoxicity and has a good clinical application value.

This invention relates to the fields of biology and biotechnology, and discloses a method for screening thermostability mutation sites in G protein-coupled receptors (GPP-CRRs) and its applications. The method includes: S1, replacing the third intracellular loop region in a wild-type GPP-CRR with the third intracellular loop sequence of a kappa opioid receptor; and labeling the wild-type GPP-CRR with an SmBiT peptide tag; S2, mutating the wild-type GPP-CRR according to the mutation sites in the GPP-CRR mutant to be tested; S3, incubating the modified GPP-CRRs from step S2 at different temperatures to obtain modified GPP-CRR samples treated at different temperatures; S4, mixing and incubating the modified GPP-CRR samples treated at different temperatures with a nanobody-LgBiT complex, then adding a luciferase substrate for detection. This invention offers high screening efficiency, low cost, and short time.

The invention provides an IGF2BP3 truncated protein mutant, an overexpression vector and application of the IGF2BP3 truncated protein mutant and belongs to the technical field of genetic engineering, and the IGF2BP3 truncated protein mutant is any one of the following items: 1) a truncated protein mutant I obtained by truncating a KH1-4 structural domain on the basis of IGF2BP3 wild type protein; 2) a truncated protein mutant II obtained by truncating a KH1-2 structural domain on the basis of IGF2BP3 wild type protein; 3) a truncated protein mutant III obtained by truncating a KH3-4 structural domain on the basis of IGF2BP3 wild-type protein.According to the invention, the specific KH structural domain can be systematically disassembled and studied through the obtained different IGF2BP3 truncated protein mutants, and further understanding of the functional mechanism of IGF2BP3 is promoted.

The invention belongs to the technical field of biomolecules, and provides application of CEL-III protein and a mutant thereof in nanopore detection. The CEL-III protein is applied to nanopore detection, has the advantages of good nanopore size uniformity, high stability, strong controllability, low cost and the like, has unique structural stability, controllability and biocompatibility obviously superior to those of an existing nanopore material, provides a new core material for a single-molecule detection technology, and has wide application prospects. The method has wide application prospects in the fields of environment monitoring, clinical diagnosis, drug screening and the like. Charge residues or hydrophobic groups are introduced into the CEL-III protein mutant through amino acid mutation, and the capture efficiency of anionic molecules and hydrophobic molecules is specifically improved; the CEL-III protein mutant can improve the quality of a current signal in nanopore detection application, and compared with wild type CEL-III protein, generated current is stable in property, the width of the current signal is narrower, burrs are fewer, and the recognition efficiency and the detection performance are improved.

The invention relates to the field of enzyme engineering, in particular to a Cry1Ie5 protein mutant with improved insecticidal activity and application of the Cry1Ie5 protein mutant. More specifically, the invention provides a Bt protein Cry1Ie5 mutant capable of improving insecticidal activity, and the Bt protein Cry1Ie5 mutant is obtained by amino acid site mutation (especially obtained by performing single-point mutation on the 50th site, the 101th site, the 111th site, the 147th site and the 550th site of an amino acid sequence of wild type Cry1Ie5 protein), so that the insecticidal activity of the Cry1Ie5 protein on corn borers is improved, and the Bt protein Cry1Ie5 mutant can be used for improving the insecticidal activity of the corn borers. And candidate genes are provided for plant transgenic insect-resistant breeding and construction of engineering strains.

The application relates to the technical field of plant breeding, and particularly relates to application of an SLR1 protein mutant or a coding gene thereof in regulating population yield and harvest index of plants. The SLR1 protein mutant is obtained by mutating at least one of tyrosines at positions 94 and 580 into alanine on the basis of a wild-type SLR1 protein. The application comprises the following steps: improving the function of the SLR1 protein mutant or the expression level of a coding gene thereof for the plants, and improving the plant type, lodging resistance, yield or harvest index of the plants. A SLR1 protein (a member of the DELLA protein family) mutant is obtained through the research of the application, and when the SLR1 protein in the plants is mutated, a semi-dwarf plant type can be obtained, the ammonium nitrogen utilization rate, the nitrate nitrogen utilization rate and the biomass are not greatly affected, the yield and the harvest index are significantly improved, and the application is superior to an sd1 green revolution variety, which has important application value in the field of plant breeding.