61 results about "Mutated protein" patented technology

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.

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 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.

This application discloses an IL-2 variant fusion protein and its applications. The IL-2 variant fusion protein of this application comprises 1) wild-type hIL-2 or an hIL-2 mutant protein, 2) an hIgG1-HC-Fc mutant fragment, and 3) an hIgG1-LC fragment; the hIL-2 mutant protein is obtained by an N88D mutation in wild-type hIL-2; the hIgG1-HC-Fc mutant fragment is obtained by L236A, L237A, and P331G mutations in wild-type hIgG1-HC-Fc. This IL-2 variant fusion protein, as a novel drug, can regulate Tregs (Foxp3). + ), treatment for Tregs (Foxp3) + This is associated with various refractory autoimmune diseases and inflammation, and prolongs the duration of Tregs (Foxp3). + Related health and lifespan span.

The application discloses a 7beta-hydroxysteroid dehydrogenase mutant and application thereof, relates to the field of bioengineering technology, and is designed and reformed according to the amino acid sequence of wild-type 7beta-hydroxysteroid dehydrogenase derived from Collinsella aerofaciens, that is, 17 single-mutation sites or 7 combined-mutation sites of 7beta-hydroxysteroid dehydrogenase are provided, the obtained 7beta-hydroxysteroid dehydrogenase mutant has the thermal stability increased by 2-21.5 DEG C and the activity increased by nearly 1.2-2.24 times compared with the wild-type 7beta-hydroxysteroid dehydrogenase. According to the mutation sites with obvious increase in thermal stability and activity, three preferred mutant proteins are further provided, the three preferred mutant proteins have higher yield, activity and thermal stability compared with the wild-type 7beta-hydroxysteroid dehydrogenase, have wider application conditions, are more suitable for the efficient production of ursodeoxycholic acid by the bioconversion method, and are favorable for large-scale production and industrial application.

This disclosure relates to novel nanoporous proteins and their applications in sequencing, belonging to the field of nanoporous technology. The nanoporous proteins provided in this disclosure enable the observation of resolvable DNA perforation signals, can be used for nanoporous sequencing, and enrich the variety of nanoporous proteins. The mutant proteins obtained after modification show significantly increased expression levels, a marked improvement in spontaneous blockage, and solve the problem of difficulty in assembling into bilayer membranes. They exhibit uniform electrophysiological properties, stable pore currents, relatively high sequencing accuracy of DNA perforation signals, and outstanding sequencing current properties.

The invention provides for a screening assay-using a selective proteolysis of recombinant full-length PIKfyve at an endogenous caspase 3 cleavage site to liberate the 34 kDa C -terminal kinase domain. Despite instability of the kinase domain to isolation, the liberated kinase domain is stable in the presence of the N-terminal PIKfyve protein fragment and can thus be treated with compounds of interest and analyzed by mass spectrometry, such as RapidFire-MS, with each injection taking under one minute. The assay was established to triage small molecules for covalent modification of PIKfyve. The assay was also established using cysteine-mutant proteins, where the reactive cysteine was replaced with an inert amino acid which when run through the same experimental protocol, was used to confirm adduct formation at the target cysteine.

Provided are a method and apparatus for recommending a protein mutation, and a computer device and a storage medium. The method comprises: receiving a mutation task request, wherein the request carries a specified first mutation instruction; in response to the request, generating third mutation instructions on the basis of the first mutation instruction and second mutation instructions; mutating an original protein sequence, so as to obtain mutated protein sequences corresponding to the third mutation instructions on a one-to-one basis; then, displaying scoring results and the third mutation instructions in a manner of corresponding to each other on a one-to-one basis; and determining whether a new mutation task request has been received, and if so, returning to continue to receive the new mutation task request. It can be seen that after the scoring results and the third mutation instructions are displayed in a manner of corresponding to each other on a one-to-one basis and a user performs experimental verification, a first mutation instruction is specified in a mutation task request again, thus forming a cyclic operation until the user finds an optimal protein sequence. Therefore, the user does not need to manually input a protein sequence, and thus the operation is convenient, and the efficiency of mutated-protein screening can be improved.

The application provides a method and system for predicting the influence of missense mutations on clinical drug efficacy based on deep learning, comprising the following steps: S1, obtaining and preprocessing drug and mutant protein sequence data sets; S2, normalizing drug input data; S3, normalizing mutant protein sequence input data; S4, building a deep learning model according to the normalized data, and outputting a prediction classification. The application can predict the influence of mutations on clinical drug efficacy only using the structure attributes of drugs and mutant protein sequences. Since the application uses a relatively small feature dimension, it supports high-throughput mutation drug efficacy prediction with low computational overhead. The application can use a deep learning method to predict the influence of mutations on clinical drug use with high precision while maintaining interpretability, and the input features do not include clinical drug information and crystal structure determination experimental information.

The present invention relates to a method for specifically degrading KRAS mutant (G12V) by treating a cell line expressing KRAS mutant (G12V) with a complex of a RAS binder that specifically binds to KRAS mutant (G12V) and an E3 ligase (E3 ligase) NEDD4 HECT, and a composition for use in the method. It can be confirmed that the method developed in the present invention, when treating a cell line expressing KRAS wild type, KRAS wild type is not degraded, in a KRAS mutant (G12V) cell line, the protein level is decreased, the ERK activity is inhibited, and the proliferation, penetration of cancer cells related to KRAS mutant (G12V) is inhibited.

This invention relates to an HSV-2 nanoparticle vaccine, its preparation method, and its application. The nanoparticles comprise HSV-2 PreF gB mutant protein and ferritin. After assembly, multiple PreF gB mutant proteins are concentrated and displayed on the surface, forming ferritin-PreF gB nanoparticles. The mutation of PreF gB enhances the stability and effective immunogenicity of the gB protein, and its display on the nanoparticle surface further enhances immunogenicity, effectively stimulating the body's cellular immune mechanism, thus becoming an effective vaccine for preventing HSV infection.

This invention discloses a Cas mutant protein and its applications, belonging to the field of gene editing technology. The Cas mutant protein provided by this invention is a mutant protein in which amino acids 168, 273, 332, 478, and 599 of SEQ ID NO:1 are all mutated to arginine, while the other amino acid sequences remain unchanged. This invention improves editing efficiency by mutating multiple amino acids in the wild-type Cas12i protein, and can be used for multi-gene editing.

The application discloses an IL12 mutant fusion protein and application thereof. Specifically disclosed are a fusion protein comprising an antibody binding to human PD1 and an IL12 mutant protein and a preparation method and therapeutic use thereof, wherein the IL12 mutant protein is obtained by mutation and modification of a wild-type P40 subunit. The fusion protein of the application has reduced binding capacity to an IL12 receptor, reduced drug toxicity of IL12, and obviously improved drugability of a cytokine. Meanwhile, through the specificity of the antibody, the fusion protein effectively enhances the targeting of the cytokine, weakens the potential toxicity of the cytokine, can exert the dual functions of the PD1 antibody and the IL12 cytokine, and thus has better anti-tumor potential. The fusion protein of the application can obviously inhibit tumor growth, has high anti-tumor effect, can prolong the half-life, reduce the administration frequency, has high safety, and has good clinical transformation value and broad application prospect.

This invention belongs to the field of biopharmaceutical technology, specifically relating to a modular protein delivery system for targeted intracellular delivery of sustained-activation cGAS protein and its applications. The modular protein delivery system comprises a first functional module and a second functional module: the first and second functional modules are connected via a fragmentation-type intron or a Cage-Colockr system; the first functional module includes the cholera exotoxin B subunit or a functionally conserved variant thereof; the second functional module includes a sustained-activation cGAS mutant protein or a lysosomal degradation-resistant derivative thereof. The modular protein delivery system provided by this invention can be used to prepare immunotherapeutic drugs for the prevention or treatment of tumors, exhibiting advantages such as strong targeting and significant immune activation effects.

The present invention relates to a population of small extracellular vesicles (sEVs) for use in the treatment of obesity in a subject in need thereof, wherein the sEVs comprise at least one polynucleotide encoding a D168A dominant negative AMP-activated protein kinase alpha 1 (AMPKa1-DN) mutant protein operably linked and under the control of a steroidogenic factor 1 (SF1) promoter, wherein the sEVs are engineered to transiently express in their outer membrane at least one fusion protein comprising the neurotrophic rabies virus (RVG) peptide fused to lysosome-associated membrane protein 2b. Said population is highly safe and effective, as the sEVs, when administered systematically, are capable of exerting their effect in the SF1 expressing neurons located in the ventromedial nucleus of the hypothalamus.

The application discloses a general trimeric recombinant protein vaccine for preventing novel coronavirus infection. Specifically disclosed are a mutant protein RBD4M 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 protective effect, can produce high-titer neutralizing antibodies against various epidemic strains of the novel coronavirus, is a broad-spectrum effective novel coronavirus vaccine, and has important significance for preventing novel coronavirus infection and a wide clinical application prospect.

This invention discloses a radiotracer targeting pan-KRAS mutant proteins, its preparation method, and its applications. The tracer uses small molecule ligands that specifically recognize multiple KRAS mutant proteins as targeting modules, and is coupled to diagnostic radionuclides through an optimized linkage system. This tracer is independent of specific mutation sites and can broadly and specifically bind to multiple high-frequency KRAS mutant subtypes, including G12C, G12D, and G12V. Based on PET / SPECT imaging, this tracer enables rapid, non-invasive, systemic, visualized, and quantitative assessment of KRAS mutant protein expression load in living tumors, overcoming the invasiveness, spatiotemporal limitations, and tumor heterogeneity challenges of traditional biopsies. It provides a novel molecular imaging tool for accurate companion diagnosis of KRAS-mutant tumors, screening of patients benefiting from pan-KRAS inhibitors, and monitoring efficacy, possessing significant clinical translational value.

This invention provides a method for optimizing a protein DDG prediction model, a protein DDG prediction method, and related apparatus, relating to the field of bioinformatics data processing technology. The method includes: acquiring wild-type protein three-dimensional structure data, mutant protein three-dimensional structure data, and a first DDG value. The wild-type protein three-dimensional structure data and the mutant protein three-dimensional structure data are respectively input into an optimized encoder in an initial protein DDG prediction model to obtain a first feature vector corresponding to the wild-type protein three-dimensional structure data and a second feature vector corresponding to the mutant protein three-dimensional structure data. Based on a variable quantum circuit, a second DDG value is obtained according to the first and second feature vectors. The initial protein DDG prediction model is iteratively optimized based on the first and second DDG values ​​to obtain a protein DDG prediction model. This invention can effectively improve the prediction accuracy of protein DDG.

This invention relates to the field of agricultural technology, specifically to a mutant Arabidopsis protein and gene that promotes chloroplast development, and their applications. The mutant Arabidopsis protein is the AtMurE protein in wild-type Arabidopsis Sig6 factor-deficient mutants, with mutations occurring at positions 89, 97, 106, 125, 126, 130, or 140. The mutant protein or gene provided by this invention, when introduced into wild-type Arabidopsis Sig6 factor-deficient mutants, can restore chlorophyll content and resolve the cotyledon yellowing problem in wild-type Arabidopsis Sig6 factor-deficient mutants.

The present application relates to the field of biological medicine, in particular to a mutant protein related to Notch signal pathway and use thereof. The mutant protein provided by the present application is a mutant Notch1 protein, a mutant DLL4 protein or a mutant JAG1 protein. The mutant protein related to Notch signal pathway provided by the present application: the mutant Notch1 protein has strong activation signal after binding with its ligands DLL4 and JAG1, and is used for regulating the disorder of Notch signal pathway, and can provide a basis for the occurrence mechanism research and development of treatment strategies of developmental disorders, cancers and other diseases.

This invention relates to the field of biodetection technology, providing a logic-gated nanogel probe for live cell typing and its preparation method. The invention uses a thermosensitive polymer nanogel as a carrier, with its surface covalently modified with an antibody that specifically binds to the EGFR L858R mutant protein and a peptide nucleic acid that specifically binds to miR-21. Internally, it encapsulates a self-quenched fluorescent dye. The probe employs an AND logic gating design, triggering gel network dissociation and dye release to generate a fluorescent signal only when it simultaneously recognizes the EGFR L858R protein on the cell surface and captures miR-21 within the cell. This invention solves the problems of high false positive rates in existing single-target detection methods and the inability of traditional liquid biopsy techniques to perform simultaneous genotype-phenotype analysis in situ on live cells. It achieves highly specific and sensitive in situ interpretation of circulating tumor cells, and has significant application value in the field of precision diagnosis and treatment of lung cancer.

The application discloses a mutant DSP12 protein and a coding gene and application thereof, and belongs to the technical field of crop genetic improvement. The application first clones a barley DSP12 gene, and the nucleotide sequence of the gene is shown as SEQ ID NO:1. Through post-mutagenesis screening of the barley, five independent functional mutation sites on the DSP12 gene are obtained, and the five independent functional mutation sites are CA / TT at the 434-435th positions, A / T at the 1022th position, G / A at the 1023th position, G / A at the 1344th position and C / T at the 1865th position, and the corresponding mutants. Field test proves that the mutants have a significantly reduced plant height, the ear type is changed from hanging to upright, and the ear density is increased, and the mutants exhibit excellent plant type structure and lodging resistance potential. The gene and the mutant DSP12 protein can be used for creating a semi-dwarf, upright and dense-ear plant type barley, and provide valuable gene resources and a new method for barley plant type improvement.

The application provides a genetically engineered bacterium for producing recovery promoting factor Rpf and application thereof, relates to the technical field of bioengineering, and belongs to prokaryotes. The nucleotide sequence of the genetically engineered bacterium is shown in SEQ ID NO. 1. The sequence is obtained by codon mutation and DNA sequence optimization of a wild-type bacterium recovery promoting factor of Micrococcus luteus in an original nucleotide sequence, and the optimized nucleotide sequence is named Delta Rpf. The original nucleotide sequence is shown in SEQ ID NO. 2. The purity of the Delta Rpf recombinant protein of the application can reach 93 %, which is much higher than the purity of the protein extracted by the prior art. The enzymatic activity of the Delta Rpf recombinant protein prepared by the genetically engineered bacterium constructed by the application is significantly higher than the enzymatic activity of the unmutated Rpf protein.

This invention belongs to the field of plant biotechnology, specifically relating to the application of the OsFTIP9 mutant in the rice seed germination process. This invention provides the application of the OsFTIP9 gene or its loss-of-function mutants in regulating the germination period of rice, wherein the mutants include those with nucleotide sequences as shown in SEQ ID No. 2~SEQ ID No. 5. This invention also provides biological materials containing the mutants, such as expression cassettes, recombinant vectors (based on pCAMBIA1300), and recombinant bacteria (such as Agrobacterium EHA105). Furthermore, this invention provides a method for regulating the germination period of rice, by transfecting the biological materials into rice tissues (such as Nipponbare) and screening for rice plants expressing the mutant protein. Experiments show that, compared with the wild type, the seed germination of the OsFTIP9 mutant is significantly delayed or inhibited. This invention provides key gene resources and effective technical solutions for elucidating the rice germination mechanism and breeding rice varieties with specific germination characteristics.

The present application relates to a VPR mutant protein capable of activating gene expression of saccharomyces cerevisiae, which is obtained by randomly mutating the commonly used activation protein VPR in saccharomyces cerevisiae, using the designed fluorescent protein and resistance gene as a screening marker, has higher transcription activation efficiency, and expands the application range of CRISPR technology in the field of gene regulation of saccharomyces cerevisiae.

This invention discloses a Bacillus subtilis lipase A mutant and its applications. Using wild-type Bacillus subtilis lipase A from existing literature as the research object, its amino acid sequence was designed and modified to obtain two mutant proteins, named BsLipA-M1 and BsLipA-M2, respectively. The Tm values ​​of BsLipA-M1 and BsLipA-M2 are 68.1℃ and 69.8℃, respectively, which are higher than those of WT (53.7℃), 20G6 mutant (49.4℃), and 5F12 mutant (60.1℃). Their activities are 1.30-2.75 times higher than the reported activities of 20G6 and 5F12 mutants. Therefore, the BsLipA-M1 and BsLipA-M2 mutants have significantly improved thermal stability and enzyme activity, which is more conducive to broadening their application range in oil and fat hydrolysis, enzymatic synthesis of biodiesel, or modification of food-grade oils.

The application belongs to the field of rice genetic breeding, and discloses a rice SBEIIb mutant protein and application thereof in changing starch crystal structure and increasing resistant starch content. The mutant protein simultaneously contains double-amino-acid substitution (M610I / D611N) of substitution of M at the 610th position to I and substitution of D at the 611th position to N. The mutant protein can change rice starch from A-type to C B -type, significantly increase amylose content from 13.8% of the wild type to 20.3%, and significantly increase starch gelatinization temperature. Meanwhile, the mutant protein (M610I / D611N) can increase the resistant starch content in rice by about 5 times; compared with the ae mutant with SBEIIb deletion, the grain width, grain thickness and grain weight are significantly increased, and the chalky kernel rate, chalky area and chalkiness are significantly reduced. The SBEIIb-M610I / D611N in the application has important application value in cultivating functional rice varieties rich in resistant starch.

The present application belongs to the field of nucleic acid editing, in particular, the field of clustered regularly interspaced short palindromic repeats (CRISPR) technology. Specifically, the present application provides a Cas mutant protein with improved activity and increased editing efficiency. Compared with the wild-type parent Cas protein, the Cas mutant protein of the present application has significantly improved activity and has a wide application prospect.