29 results about "New mutation" patented technology

The invention relates to a gene diagnosis technology, in particular to a kit for detecting phenylketonuria pathogenic genes and application of the kit, the kit comprises a reagent for detecting PAH pathogenic mutants, and the PAH pathogenic mutants are characterized in that in a coding region of a nucleotide sequence shown as the accession number NM000277.3, the 721st basic group is mutated into C from T, and the 741st basic group is mutated into G from T; the reagent is a primer. According to the invention, a new mutation site of the phenylketonuria virulence gene is found, and the research of the phenylketonuria virulence gene is enriched; the kit provided by the invention contains a reagent for detecting the new mutation site, is found for the first time in the prior art, is simple in structure, can more conveniently and quickly detect the pathogenic gene of phenylketonuria, and increases a new clinical detection product.

The present invention provides a class of novel mutant proteins for increasing the malic acid yield. Specifically, the present invention provides a class of novel pyruvate carboxylase mutant proteins and malic acid transporter mutant proteins or combinations thereof, as well as methods for their preparation and applications in increasing the malic acid yield.

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 present invention offers a new approach for highly multiplexed, programmable antiviral therapies that directly target viral RNA, and can be flexibly adapted to target novel viruses or emerging outbreak pathogens. Class 2, type VI CRISPR system-based therapies can be used in combination with existing antiviral compounds for viruses where such compounds exist, either by increasing their efficacy or by preventing the evolution of specific drug resistance mutations. Perhaps most excitingly, if a virus evolves resistance to a specific guide RNA sequence, it is easy to switch to a different guide RNA sequence, or to design a new guide sequence to target the new mutation. Such approaches should prevent the widespread development of resistance to Class 2, type VI CRISPR system-based therapies.

The invention relates to the technical field of cell engineering, in particular to a cell strain causing pathogenic splicing mutation of a female HA patient and a construction method of the cell strain. A construction method of the T point mutation cell strain comprises the following steps: designing an sgRNA sequence based on a mutation site of a target gene F8; the method comprises the following steps: based on an sgRNA sequence and a CRISPR / Cas9 gene editing technology, constructing a plurality of pX458-sgRNA plasmids by taking a pX458 plasmid as a vector; the method comprises the following steps: constructing an LA-pMD-18T-pGK-puro-mcherry-RA donor plasmid by taking a pMD-18T-pGK-puro-mcherry plasmid as a vector on the basis of an sgRNA (small guide ribonucleic acid) sequence and a pX458-sgRNA plasmid; the preparation method comprises the following steps: simultaneously transfecting a pX458-sgRNA plasmid and an LA-pMD-18T-pGK-puro-mcherry-RA donor plasmid into a cell, and carrying out drug screening, so as to obtain a stable cell strain. In addition, the invention also provides application of the mutant cell strain. The discovery of the novel pathogenic splicing mutation not only enriches an F8 gene mutation database, but also provides a new strategy for precise treatment of female HA patients in the future.

The invention relates to a Sanger sequencing method primer group and a kit for human RHD blood group genotyping detection, and belongs to the field of biomedicine clinical molecular detection. The Sanger sequencing method primer group for human RHD blood type genotyping detection comprises a specific PCR (Polymerase Chain Reaction) amplification primer group, and the specific PCR amplification primer group comprises 9 pairs of amplification primers designed according to an RHD gene specific sequence; and the primer group also comprises 10 RHD specific sequencing primers corresponding to the amplification products. The RHD gene specific amplification primer has the following technical effects that the RHD gene specific amplification primer designed by adopting an ARMS combined homologous sequence specific base method is higher in specificity and accuracy; in cooperation with the RHD specific sequencing primer, the gene sequence of a sample to be detected is detected through a Sanger sequencing method, the RHD genotype is interpreted, and a new mutation point can be found. According to the kit disclosed by the invention, the identification results of the exons from the RHD blood group genes 1 to 10 can be obtained, so that the RHD genetic typing of an experimental sample can be accurately judged.

The present invention relates to a subtilisin BPN' variant or homologue thereof comprising new mutations compared to subtilisin BPN' as shown in SEQ ID NO: 2 or a homologous sequence thereof. Such mutations can occur at amino acid positions selected from the group consisting of L96, D99, A223 and S224. The present invention also relates to a method for the enzymatic synthesis of a peptide by coupling of peptide fragments, wherein said coupling is catalysed by said subtilisin BPN' variant or homologue thereof.

The present invention relates to a subtilisin BPN'variant or a homolog thereof comprising a new mutation compared to a subtilisin BPN 'as shown in SEQ ID NO: 3 or a homologous sequence thereof. This mutation may occur at an amino acid position selected from the group consisting of H17W, S18K, N25G, V30I, S63G, T71V, V72I, S204N, E251L, N43K or S87D.

The application discloses a test case automatic generation method and system based on MoMuTUML, reduces and deletes original function overlapping mutation operators, and adds three new mutation operators which are not functionally overlapped with the original mutation operators, so that the generation of a syntax equivalent mutation model is reduced, and the test case generation time cost is reduced; test cases are generated according to a mutation model set and a model under test, and equivalent mutation models are deleted according to the test cases, so that effective mutation models are obtained, the problem of too many equivalent mutation models in MoMuTUML is avoided, and the test case generation efficiency is improved; through a feedback-based mutation method, the mutation operators corresponding to the effective mutation models are scored, an optimal mutation operator set is constructed, and the mutation operators in the set are selected to perform mutation operations on the model under test, so that the problem of low efficiency of the MoMuTUML mutation method is solved, and the test case generation efficiency is further improved.

The invention relates to the technical field of plant genetic engineering, and discloses a rice TMS5 gene mutant and application thereof in rice thermo-sensitive sterility, the mutant is formed by replacing a 176th base of a rice TMS5 gene coding region with a C base, the nucleotide sequence of the TMS5 gene mutant is shown as SEQ ID No.1, and the amino acid sequence of the TMS5 gene mutant is shown as SEQ ID No.2. The mutation site and the mutation mode of the mutant are different from those of an existing reported TMS5 gene mutant, and the mutant is a new mutant allelic of the TMS5 gene, shows a typical temperature-sensitive sterile phenotype, does not generate pollen at a high temperature, can generate pollen at a low temperature, and has huge potential in the application of rice crossbreeding.

The present invention provides a new mutant protein for increasing malic acid production. Specifically, the present invention provides a new pyruvate carboxylase mutant protein and a malic acid transporter mutant protein or a combination thereof, as well as a preparation method thereof and an application thereof in increasing malic acid production.

The invention provides a novel base editing system for mediating multi-base saturation mutation. Specifically, the present invention provides a multi-base editor comprising one or more elements selected from the group consisting of: (a) a nuclear localization signal; (b) a cytosine deaminase; (c) adenine deaminase; (d) a Cas protein or a variant thereof or an active fragment thereof; (e) an alkyl adenine DNA glycosylase or a variant thereof; (f) a linker; and optionally (g) an N-methylpurine DNA glycosylase protein. The invention also provides a multi-base editing system based on the multi-base editor, a corresponding polynucleotide, a vector, a host cell and application thereof. According to the multi-base editor and the editing system disclosed by the invention, saturation mutation of two bases and three bases (mediating Agt; c / T / G and / or Cgt; t / G / A and / or Ggt; c / T / A), in particular, a new mutation type Agt is generated; c / Tamp; cgt; t / G / A and Agt; c / T / Gamp; cgt; t / G / Aamp; ggt; the rich saturated mutation spectrum of C / T / A can greatly promote gene function screening, protein evolution, pedigree tracing and other applications.

The invention discloses a viral genome data analysis and prediction system based on deep learning, and relates to the technical field of data management. Through a deep learning model module, a convolutional neural network is utilized to extract genome local features and realize classification and variation detection; the robustness evaluation module quantifies the performance fluctuation of the model in new mutation by analyzing a new mutation test data set; a confidence coefficient analysis module evaluates abnormity of confidence coefficient distribution predicted by the model, a comprehensive analysis module combines robustness and a confidence coefficient result to generate a comprehensive stability score, and the stability of the model to new mutation is dynamically quantified; and the model optimization module iteratively updates a model architecture and a training data set based on an analysis result, so that the prediction performance of the model on the dynamic variation of the virus is remarkably improved, the problem that the existing model is difficult to adapt to the accumulated drift of the virus genome mutation is effectively solved, an efficient and stable solution is provided for rapid detection of new mutation, and the method is suitable for popularization and application. Important public health significance is realized.

The present application relates to a kind of primer set and kit for Sanger sequencing method for human RHD blood group genotyping detection, belong to biomedical clinical molecular detection field.The primer set for Sanger sequencing method for human RHD blood group genotyping detection of the present application, including specific PCR amplification primer set, the specific PCR amplification primer set includes 9 pairs of amplification primer designed according to RHD gene specific sequence;The primer set also includes 10 RHD specific sequencing primers corresponding to amplification product.The present application has the following technical effects: the present application adopts the RHD gene specific amplification primer designed by ARMS and homologous sequence specific base method, and specificity and accuracy are higher;With the RHD specific sequencing primer of the present application, the gene sequence of the sample to be detected is detected by Sanger sequencing method, RHD genotype is interpreted, and new mutation point can be found.The identification result of RHD blood group gene 1 to 10 exons can be obtained by the kit of the present application, so as to accurately determine the RHD gene typing of experimental sample.

The invention discloses a probe set for evaluating whole-body tumor load of small cell lung cancer and application of the probe set, the probe set comprises a plurality of oligonucleotide probes for specifically capturing mutation regions of SCLC related genes in a targeted mode, and the genes are selected from TP53, RB1, CREBBP, EGFR, PTEN, PIK3CA, LRP1B and NOTCH1; and the probe set is capable of covering at least 94.9% of at least one mutation existing in an SCLC patient sample. The invention relates to the technical field of biotechnology and molecular diagnosis, and has the beneficial effects that based on a clinical queue, the probe group incorporates high-frequency mutant genes of SCLC, can be superposed with personalized probes for use, and is used for ctDNA detection of SCLC patients and evaluation of whole-body tumor load. The SCLC cancer species specific probe group provided by the invention plays a role in monitoring tumor evolution and new mutation, can overcome the space-time heterogeneity of tumors to a certain extent, and also can improve the capture efficiency at the same time.

The application belongs to the technical field of gene detection, and particularly relates to a construction method and kit of a gene library for detecting new mutations of dominant monogenic diseases before embryo implantation. The application provides a construction method of a gene library for detecting new mutations of dominant monogenic diseases before embryo implantation, which comprises the step of amplifying the gene of the new mutation of dominant monogenic diseases before embryo implantation and related copy number variation fragments in the genomic DNA of specific screening. The application precisely applies NGS sequencing technology to the screening of dominant monogenic diseases in trace cell samples, breaks through the technical bottleneck of high-throughput sequencing under extremely low initial DNA amount in the past, and realizes the screening of new mutations before embryo implantation, thereby providing a brand-new key technical path for blocking genetic birth defects at an earlier stage.

The application discloses a virus genome data analysis and prediction system based on deep learning, relates to the technical field of data management, and extracts local features of a genome and realizes classification and variation detection by using a convolutional neural network through a deep learning model module; a robustness evaluation module quantifies the performance fluctuation of the model in new mutations by analyzing a new mutation test data set; a confidence analysis module evaluates the abnormality of the model prediction confidence distribution; a comprehensive analysis module generates a comprehensive stability score in combination with the robustness and confidence results, dynamically quantifies the stability of the model to new mutations; and a model optimization module iteratively updates the model architecture and training data set based on the analysis results, thereby significantly improving the prediction performance of the model on dynamic variation of viruses, effectively solving the problem that existing models are difficult to adapt to the cumulative drift of virus genome mutations, providing an efficient and stable solution for rapid detection of new mutations, and having important public health significance.

The invention relates to a human RHCE blood type genotyping primer group based on Sanger sequencing and a kit, and belongs to the field of biomedicine clinical molecular detection. The primer group disclosed by the invention comprises a specific PCR (Polymerase Chain Reaction) amplification primer group, wherein the specific PCR amplification primer group comprises 9 pairs of primers designed according to a RHCE gene specific sequence; the 9 pairs of primers are respectively used for amplifying RHCE exons 1 to 10. And the primer group also comprises 10 RHCE (Recombinant Heterogeneous Cell Element) specific sequencing primers corresponding to the amplification primers. Compared with the prior art, the invention has the following technical effects: the specificity of the specific amplification primer designed by combining ARMS with a homologous sequence specific base method is high, the gene sequence of a sample to be detected is detected by matching with the RHCE specific sequencing primer, the RHCE genotype is interpreted by a Sanger sequencing method, and a new mutation point can be found. According to the kit disclosed by the invention, the identification result of the RHCE blood group gene exons 1-10 can be obtained, so that the RHCE high-resolution genetic typing of an experimental sample can be accurately judged.

The present invention belongs to the field of gene technology, and in particular to KIF11 gene mutants and applications. Specifically disclosed is a nucleic acid comprising the following target fragment, wherein the target fragment has a c.2680C>T mutation compared with the wild-type KIF11 gene. A polypeptide, wherein the polypeptide has a p.Gln894Ter mutation compared with the wild-type KIF11. A gene mutation, wherein the gene mutation has a c.2680C>T mutation compared with the wild-type KIF11 gene. Application of a biological model containing the aforementioned mutation in the preparation of a screening preparation for microcephaly, without chorioretinopathy, lymphedema, or mental retardation. A screening reagent for microcephaly, without chorioretinopathy, lymphedema, or mental retardation, comprising a reagent capable of detecting KIF11 gene mutants. The present invention provides new mutation sites associated with microcephaly, without chorioretinopathy, lymphedema, or mental retardation, provides a new scheme for the diagnosis and treatment of the disease, and also provides related drugs capable of preventing and treating the disease.

This invention discloses a BTD gene knockdown hepatocyte injury model and its construction method, relating to the fields of molecular biology and cell biology. The model is used to knock down the expression of the BTD gene in human hepatocytes to construct a hepatocyte injury model. Its sense strand nucleotide sequence is 5'-GCGAUUGGUCUCAAGCUAA(dT)(dT)-3', and its antisense strand nucleotide sequence is 5'-UUAGCUUGAGACCAAUCGC(dT)(dT)-3'. This invention designs specific siRNA sequences to directionally knock out the BTD gene in human hepatocytes, thereby observing and verifying whether it leads to abnormalities in liver injury indicators. It clarifies the direct causal relationship between BTD gene functional defects and hepatocyte injury, providing powerful experimental tools and data support for the gene diagnosis of unexplained liver diseases, the pathogenicity assessment of new BTD gene mutation sites, and the development of related drugs.

The invention belongs to the technical field of molecular detection, and particularly relates to a GluCl2 subunit mutation site-based detection reagent for drug resistance of spider mites to abamectin and a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) detection method. The GluCl gene sequence in a tetranychus urticae population collected from a field is analyzed to find that a new mutation site T76S exists on a GluCl2 subunit, the mutation exists in a field population with multiple abamectin resistance, and the mutation frequency of the site is increased after abamectin treatment. In the process of identifying the mutation site, the mutation site is identified; according to the present invention, the specific recognition site (T 'CAG) of the DdeI restriction enzyme in the sequence is identified, such that the amplification primer is designed, the PCR-RFLP method for T76S mutation site detection is established, and the method has characteristics of rapid enzyme digestion reaction, simple operation, pollution resistance, short time, accurate experiment result, and easy determination.

This invention is a one-dimensional material cutting method based on gene evaluation genetic algorithm. By randomly combining gene materials, the composition of each gene can be intuitively understood, and the cutting of each steel bar can also be known. The time spent on decoding is reduced. By scoring each gene in the chromosome, excellent genes are selected for mutation, and the optimal solution can be obtained more easily and quickly. The new mutation method reduces the evaluation time of individuals and speeds up the convergence speed of the algorithm.

The invention relates to a subtilisin BPN′ variant or homologue thereof, having new mutations compared to subtilisin BPN′ represented by SEQ ID NO: 2 or a homologue sequence thereof. Such mutations may occur at amino acid positions L96, D99, A223 or S224. The invention further relates to a method for enzymatically synthesizing a peptide by coupling peptide fragments, wherein the coupling is catalyzed by the subtilisin BPN′ variant or homologue thereof.

The present application relates to a kind of human RHCE blood group genotyping primer group and kit based on Sanger sequencing, belong to biomedical clinical molecular detection field.The primer group of the present application includes 9 pairs of primers designed according to the specific sequence of RHCE gene;The 9 pairs of primers are used to amplify RHCE 1 to 10 exons.The primer group also includes 10 RHCE specific sequencing primers corresponding to amplification primer.The specific amplification primer designed by ARMS combined with homologous sequence specific base method is high in specificity, cooperates with the RHCE specific sequencing primer of the present application, detects the gene sequence of the sample to be detected by Sanger sequencing method, interprets the RHCE genotype, and new mutation point can be found.The identification result of RHCE blood group gene 1 to 10 exons can be obtained by the kit of the present application, so as to accurately determine the RHCE high-resolution genotyping of experimental sample.

The invention relates to promotion of breast cancer proliferation and metastasis by endogenous hydrogen sulfide mediated PGK1 sulfhydrylation modification, belongs to the technical field of breast cancer proliferation and metastasis, and aims to solve the problem that partial patients suffer from local relapse or distant metastasis due to drug resistance caused by new mutation of tumor cells. The method comprises the following steps: real-time fluorescent quantitative PCR (Polymerase Chain Reaction), Western blot, H2S content detection of a tissue sample, H2S content detection of a cell sample, Biotin switch assay, cell culture, a plasmid related experiment, a CCK-8 experiment, a plate cloning experiment, a cell scratch experiment, a Transwell migration experiment, a Transwell invasion experiment, glucose level detection, lactic acid level detection, ATP (Adenosine Triphosphate) level detection and determination of an extracellular acidification rate and an oxygen consumption rate. The method comprises the following steps: construction of a mouse breast cancer model, hematoxylin-eosin staining, immunohistochemical staining and statistical analysis, and endogenous H2S regulates glycolysis of breast cancer cells by mediating PGK1 sulfhydrylation modification so as to promote proliferation and metastasis of tumors.

The use of a soybean GmGID1-2 gene in the regulation of soybean plant architecture, nitrogen use efficiency, high yield and high oil content. The GmGID1-2 gene has a CDS sequence as shown in SEQ ID NO: 1. The knockout of the GmGID1-2 gene can improve plant architecture (reducing plant height, and increasing branch number, node number of main stem, and pod number), increase stem diameter and stem strength, enhance nitrogen fixation in root nodules, and improve yield and seed oil content. For the pleiotropic gene GmGID1-2, by means of gene editing technology, two new mutations are generated in GmGID1-2, resulting in premature termination of a protein. The knockout of the GmGID1-2 gene simultaneously realizes the improvement of multiple traits in crops such as soybean.

The application provides a method and device for recommending protein mutations, a computer device and a storage medium, comprising: receiving a mutation task request, the request carrying a specified first mutation instruction; in response to the request, generating a third mutation instruction based on the first mutation instruction and a second mutation instruction; mutating an original protein sequence to obtain a mutated protein sequence corresponding to each third mutation instruction; then displaying the score results and the third mutation instruction one by one; determining whether a new mutation task request is received; if yes, continue to receive the new mutation task request; it can be seen that the score results and the third mutation instruction are displayed one by one, and the first mutation instruction is specified in the mutation task request after the user performs experimental verification, thus forming a cyclic operation until the user finds the optimal protein sequence, so that the user does not need to manually input the protein sequence, the operation is convenient, and the screening efficiency of the mutated protein can be improved.

The invention belongs to the technical field of biology, and particularly relates to a thermal-stability single-chain DNA / RNA ligase mutant and application thereof. Compared with the amino acid sequence (pfumlinase) of SEQ ID NO: 1, the DNA / RNA ligase with the mutated lysine provided by the invention has the advantages that the 372 site is subjected to substitution mutation; the amino acid sequence of the nucleic acid ligase is as shown in SEQ ID NO: 4. According to the present invention, the potential connection active site is screened, and the gene mutation modification is performed so as to obtain the Pfumrigase DNA / RNA ligase with the new mutation site; compared with the original Pfumrigase DNA / RNA ligase, the modified Pfumrigase DNA / RNA ligase has the advantage that the modified Pfumrigase DNA / RNA ligase has higher catalytic activity.

The application belongs to the technical field of molecular detection, and particularly relates to a detection reagent for detecting the resistance of a spider mite to abamectin based on a mutation site of a GluCl2 subunit and a PCR-RFLP detection method. GluCl Gene sequence analysis finds that there is a new mutation site T76S on the GluCl2 subunit, the mutation exists in multiple abamectin-resistant field populations, and the mutation frequency of the site is increased after abamectin treatment. In the process of identifying the above mutation site, it is found that there is a specific recognition site (T'CAG) of DdeI restriction endonuclease in the sequence, and thus an amplification primer is designed, and a PCR-RFLP method for detecting the T76S mutation site is established. The method is fast in enzyme digestion reaction, simple in operation, not easy to be contaminated, short in time cost, accurate in experimental results, and easier in judgment method.