1470 results about "Mutation" patented technology

The invention discloses a bovine I-type alpha interferon-ferritin fusion protein, a mutant thereof, a preparation method and an application of the bovine I-type alpha interferon-ferritin fusion protein. The bovine I-type alpha interferon is fused with a ferritin subunit, and interferon molecules are highly repeatedly and orderly displayed on the surface of a ferritin nanocage by utilizing the self-assembly characteristic of ferritin, so that the expression level, the structural stability and the antiviral activity of the interferon are remarkably improved. The fusion protein is further subjected to single-site or multi-site rational design mutation, and a mutant with significantly improved antiviral activity and stability is obtained. According to the invention, a silkworm or insect cell eukaryotic expression system is adopted to express the fusion protein or the mutant thereof, and the expression system is safe to operate, simple and convenient in procedure, low in cost and extremely beneficial to large-scale industrial production; the prepared fusion protein or mutant nanoparticles have application prospects in preparation of drugs or reagents for preventing or treating bovine viral diseases.

The invention belongs to the technical field of pepper planting, and discloses a pepper multi-branch gene CaBr1 linked KASP molecular marker as well as a primer, a kit and application thereof, the KASP molecular marker takes pepper S8 (zhangshuang) as a reference genome, and the single nucleotide polymorphism at the 103175415th basic group on the No.8 chromosome is subjected to basic group G-to-A replacement. A chromosome region closely linked with multiple branches of the pepper is obtained by using a BSA (Bovine Serum Albumin) population positioning method, a molecular marker is developed in a candidate region to determine a candidate gene CaBr1, a KASP molecular marker is screened according to base mutation of the candidate gene, and the marker is used for carrying out genotype identification on 90 randomly sampled single plants of an F2 population, and the coincidence rate reaches 100%. The result not only contributes to pepper branch phenotype identification and assistant breeding, but also lays a foundation for map-based cloning of the branch gene and analysis of a molecular mechanism of pepper plant type regulation.

The invention discloses a rice salt stress resistant gene OsHAK11 coding protein and application thereof, and belongs to the field of plant genetic engineering. According to the method, a rice OsHAK11 gene (the nucleotide sequence is shown as SEQ ID NO.1) is knocked out through a CRISPR / Cas9 gene editing technology, and a mutant with significantly enhanced salt tolerance is obtained. The gene editing vector pEGCas9Pubi-B-OsHAK11 contains sgRNA of a region as shown in a target SEQ ID NO.4, a rice receptor material is transformed through agrobacterium tumefaciens mediation, and a plant with the OsHAK11 gene subjected to frame shift mutation is obtained through screening. A salt stress experiment shows that the survival rate of the mutant oshak11 is obviously higher than that of a wild type. The invention provides a new gene resource and an efficient technical means for salt-tolerant breeding of rice.

The invention belongs to the field of biosynthesis, and in particular relates to LBD (Levandanenediol Pyrophosphate) synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, the sclareol biosynthesis yield is generally low, LSLPPs mutant containing at least one mutation of D730L, D374L, N674L or G379M is obtained by performing mutation and optimization on LSLPPs pyrophosphate synthase, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, and the yield of a 5L fermentation tank reaches 26.11 g / L and is greatly improved compared with the prior art.

The invention belongs to the field of biosynthesis, and particularly relates to sclareol synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, sclareol biosynthesis yield is generally low, sclareol synthase is mutated and optimized, an SsScs mutant containing at least one mutation of Y491F, G307L or N269F is obtained, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, and the yield of a 5L fermentation tank reaches 21.13 g / L and is greatly improved compared with the prior art.

The invention belongs to the technical field of bioengineering, and relates to a transaminase mutant, a recombinant genetically engineered bacterium and application of the transaminase mutant in catalytic synthesis of (R)-1-Boc-3-aminopiperidine.The transaminase mutant is obtained by conducting single-point or combined mutation on the 131 site and / or the 197 site of an amino acid sequence shown in SEQ ID NO.2; the mutation sites comprise that the 131 phenylalanine is mutated into aspartic acid, threonine or tyrosine, and / or the 197 lysine is mutated into arginine or leucine. Experimental results show that compared with wild type transaminase, the catalytic activity, the thermal stability and the organic solvent tolerance of the obtained mutants, especially single-point mutants MyTA1-F131Y and MyTA1-K197R and a combined mutant MyTA1-F131Y-K197R, are all remarkably improved, and compared with the wild type transaminase, the catalytic activity, the thermal stability and the organic solvent tolerance of the obtained mutants are all remarkably improved. The mutant MyTA1-F131Y-K197R can be used for efficiently catalyzing asymmetric amination of N-Boc-3-piperidone to synthesize (R)-1-Boc-3-aminopiperidine, the conversion rate of the (R)-1-Boc-3-aminopiperidine after the (R)-1-Boc-3-aminopiperidine reacts for 24 hours under the condition that the substrate concentration is 100 g / L can reach 90% or above, and the mutant MyTA1-F131Y-K197R has a good industrial application prospect.

The invention discloses a molecular marker remarkably related to the amylose content of wheat and application of the molecular marker, and belongs to the technical field of wheat breeding. Three mutation sites of wheat starch branching enzyme gene SBEIIa on 2A, 2B and 2D chromosomes and two mutation sites of SBEIIb on 2A and 2B chromosomes are identified, a KASP molecular marker system closely linked with SBEIIa and / or SBEIIb is developed based on the SNP sites, the detection process is simple and convenient, the cost is low, the throughput is high, and the application prospect is wide. According to the method, the high-amylose wheat material which contains the mutant genotype and can be stably inherited can be accurately and efficiently screened out, the breeding process of wheat variety quality improvement can be accelerated, and the method has important significance on creation of the high-amylose wheat material or molecular assisted breeding of the high-amylose wheat.

The invention belongs to the technical field of bioinformatics, and discloses a protein disulfide bond mutation site prediction method and device, equipment and a storage medium, a plurality of candidate residue pairs are extracted by obtaining a prediction structure of a to-be-modified protein, and structural feature information of each candidate residue pair is extracted; comprising an inter-residue distance matrix of two residues, a sequence distance, a relative solvent accessible surface area of each residue, a depth index and a predicted local distance difference test value, inputting a prediction model to obtain a probability value of forming a disulfide bond by each candidate residue pair, determining the candidate residue pair with a probability greater than a specified probability as a disulfide bond mutation site, therefore, the bonding probability of the disulfide bond can be comprehensively predicted by using more comprehensive features, the high dependence of the model on individual traditional features is reduced, the high sensitivity of a traditional method on atomic coordinates and the uncertainty of a prediction result are overcome to a certain extent, and the method has higher robustness and is suitable for popularization and application. The prediction accuracy of the protein disulfide bond mutation site is improved.

The disclosure features systems and methods for correcting a mutation in the human beta-globin (HBB) gene in a cell or population of cells. The disclosure also features methods of increasing repair of a DNA double stranded break (DSB) in an HBB gene by the homology-directed repair (HDR) pathway. The disclosure also features compositions for use in the methods.

The invention provides methods, systems, and computer readable medium for detecting ploidy of chromosome segments or entire chromosomes, for detecting single nucleotide variants and for detecting both ploidy of chromosome segments and single nucleotide variants. In some aspects, the invention provides methods, systems, and computer readable medium for detecting cancer or a chromosomal abnormality in a gestating fetus.

The present disclosure relates to Taq DNA Polymerase mutants comprising at least three mutations which are capable of high levels of DNA amplification in the presence of several PCR inhibitors and compositions comprising the same. Also disclosed are methods of performing mutagenesis on wild-type Taq DNA polymerase to obtain the disclosed Taq DNA polymerase mutants, DNA amplification using the same, and cDNA amplification using the same.

Compositions, methods, kits, systems, and software are provided for non-invasive prenatal testing for autosomal recessive diseases. Next generation sequencing is used to sequence maternal and fetal DNA isolated from maternal plasma by probe capture. The fetal fraction of the sequencing reads for DNA isolated from maternal plasma is estimated by counting single nucleotide polymorphisms (SNPs) for which an allele is detected that is present in the paternal haplotype but absent in the maternal haplotype, based on the assumption that SNPs having a paternal allele belong to the fetal DNA. The fetal fraction is bioinformatically enriched by excluding sequencing reads over a specified length via in-silico size selection, which increases fetal genotype prediction accuracy. Parental haplotype information together with the read ratios observed at the linked SNPs is used to predict the fetal genotype at a site of a mutation linked to the autosomal recessive disease.

The invention discloses a VdNOB1 gene as well as an anti-pathogenic bacterium target gene segment, an interference vector and disease-resistant application thereof. According to the invention, the VdNOB1 gene is subjected to knockout mutation, so that the infection pathogenicity of verticillium dahliae to host plants can be obviously reduced. The invention further provides a verticillium dahliae VdNOB1 gene anti-pathogenic bacterium target gene segment, the nucleotide sequence of the verticillium dahliae VdNOB1 gene anti-pathogenic bacterium target gene segment is shown as SEQ ID No.4, and the pathogenicity of verticillium dahliae to host plants can be remarkably reduced after plants are transformed by an RNA interference vector constructed by the verticillium dahliae VdNOB1 gene anti-pathogenic bacterium target gene segment. The invention further provides an RNA (Ribonucleic Acid) interference vector containing the VdNOB1 gene or an anti-pathogenic bacterium target gene fragment thereof. The verticillium dahliae VdNOB1 gene or the anti-pathogenic-bacterium target gene segment and the RNA interference vector thereof have application prospects in the aspects of improving the resistance of crops to diseases caused by verticillium dahliae, cultivating new varieties of verticillium dahliae-resistant plants and the like.

The invention discloses a high-specificity minimal residual disease (MRD) monitoring method and system based on circulating tumor DNA (ctDNA). The method comprises the following steps: receiving tumor tissue sequencing data of an UTUC patient, and generating a double-Panel target list containing personalized and fixed Panel; respectively extracting plasma cfDNA and leukocyte gDNA; performing vacuum concentration, hybrid capture and sequencing on the cfDNA library by using the double Panel lists, and performing deep sequencing on the leukocyte gDNA; constructing an individualized clonal hematopoietic mutation filtering database; actively filtering and rejecting clonal hematopoietic background mutation by utilizing a filtering database; and calculating an MRD load score based on the filtered tumor-derived mutation and outputting a report. The system comprises corresponding modules which are used for automatically executing the process. According to the invention, through cooperation of four major technologies of double-Panel design, process optimization, UMI error correction and active clonal hematopoietic filtration, MRD monitoring with extremely high sensitivity and specificity on UTUC is realized, false positive is significantly reduced, and the kit has drug resistance early warning potential.

The present disclosure provides methods and systems for identifying transcriptional regulatory modules (e.g., in non-coding portions of the genome), predicting gene regulation and expression, e.g., effects of non-coding mutations or chromosome rearrangements on the regulation and expression of the target genes, and designing and using modified regulatory sequences.

The disclosure provides a method for treating a subject afflicted with a tumor derived from a small cell lung cancer (SCLC) having a high tumor mutational burden (TMB) status comprising administering to the subject a monotherapy comprising an anti-PD-1 antibody or a combination therapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody. The present disclosure also provides a method for identifying a subject suitable for treatment with an anti-PD-1 antibody or a combination therapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody comprising measuring a TMB status of a biological sample of the subject. A high TMB status identifies the patient as suitable for treatment with an anti-PD-1 antibody or antigen-binding portion thereof. The TMB status can be determined by sequencing nucleic acids in the tumor and identifying a genomic alteration, e.g., a somatic nonsynonymous mutation, in the sequenced nucleic acids.

The invention is suitable for the technical field of soybean breeding, and provides a KASP molecular marker for soybean plant height identification and application of the KASP molecular marker. The KASP molecular marker is based on an SNP variation site of a Glyma. 19G206100 gene between two parents of Chinese flat stem soybean and F02, the mutation position of the Glyma. 19G206100 gene is the 312th basic group on a CDS sequence, C of a reference genome is mutated into G, the 104th amino acid on a protein sequence is mutated into lysine from asparagine, and the 104th amino acid on the protein sequence is mutated into lysine from asparagine. The primer of the KASP molecular marker comprises a forward primer with a nucleotide sequence as shown in SEQ ID NO.1-2 and a reverse primer with a nucleotide sequence as shown in SEQ ID NO.3. The invention further discloses a kit for detecting the KASP molecular marker. The KASP molecular marker is developed on the basis of the polymorphic site of Gm19G206100 in germplasm resources, the marker has the advantages of being high in resolution ratio and easy to detect, can be applied to plant height selection of early generations of soybean cross breeding, has the effects of reducing the breeding scale, reducing the workload and improving the breeding efficiency, and has important application value in production.

The invention relates to the technical field of corn breeding, in particular to a corn dwarfing gene as well as a primer pair composition, a kit, a detection method and application thereof. The corn dwarfing gene is obtained after a Br2 gene is subjected to single base mutation, the gene number of the Br2 gene is Zm00001eb038710, the Br2 gene is br2-6, br2-7, br2-70, br2-98, br2-100, br2-113, br2-115, br2-128, br2-129, br2-130 or br2-131, and the corn dwarfing gene can be used for detecting the corn dwarfing gene. Eleven brand-new single-base mutation sites in the corn Br2 gene (the gene number is Zm00001eb038710) are defined, the mutation of the sites can effectively reduce the height of corn plants by regulating and controlling gene functions, and accurate and efficient targets are provided for corn plant type improvement and molecular assisted breeding.

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker related to disease resistance of ricefield eel and application of the SNP molecular marker, and relates to the technical field of molecular biology. The nucleotide sequence of the SNP molecular marker is as shown in SEQ ID NO.3, and SNP sites, namely G / A mutation and A / C mutation, exist at the 143rd and 413th basic groups of the SNP molecular marker. According to the invention, SNP loci in LamR genes in a ricefield eel group are screened out through sequence alignment, and genotyping is carried out on the SNP loci; and comparing the differences of the disease resistance of the genotypes, and screening to obtain the genotype with the strongest disease resistance, thereby providing an effective molecular marker for detecting the disease resistance of the ricefield eel. The SNP molecular marker provided by the invention is associated with the disease resistance of the ricefield eel, and provides effective technical support for breeding of disease-resistant ricefield eel varieties.

The invention belongs to the technical field of medicines, and provides a method for screening a mutant strain and a high-virulence Seneca virus A mutant strain. The method for promoting Senecavirus A mutation provided by the invention comprises the following steps: inserting an RNAi target sequence between 3Dpol and a 3'untranslated region of an SVA starting strain; the target sequence of the RNAi contains a complementary sequence of an endogenous sequence of the miRNA. According to the research, mutagenesis is carried out on SVA by constructing a high RNAi pressure system, and a method for screening the mutant strain SVA is carried out by simulating an antiviral RNAi mechanism in a host, so that a natural evolution process of the virus is better met, a large number of stably inherited mutant strains can be obtained, and an efficient research and development means is provided for vaccine research and development aiming at the virus.

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 technical field of gene detection, and discloses a mycobacterium tuberculosis drug resistance gene detection data analysis system which comprises a data import module, a signal correction module, a feature extraction module, a multi-stage interpretation module, a result fusion module and a report generation module. The system performs baseline drift correction, abnormal point detection and nonlinear normalization on a fluorescence intensity value output by a detector, extracts multi-dimensional features such as peak intensity, a mutation ratio, a consistency coefficient and an in-batch difference index, and constructs a dynamic threshold function to realize wild type and mutation type layered judgment. A drug resistance risk conclusion is generated through drug resistance characteristic matrix mapping, a standardized detection report is output, and intelligent analysis of a detection result and automatic report generation are achieved. Through multi-stage signal correction, multi-dimensional feature extraction and dynamic threshold interpretation, intelligent analysis and standardized report output of a mycobacterium tuberculosis drug resistance gene detection result are realized, and data judgment accuracy and clinical application efficiency are improved.

SpRYc is a grafted ScCas9++-SpRY chimeric Cas9 possessing minimal 5′-NNN-3′ PAM specificity. SpRYc comprises the N-terminus (residues 1-1119) of ScCas9++ (Sc++), including the flexible loop, followed by the region of SpRY (residues 1111-1368) spanning its PAM-interacting domain mutations. Methods of altering gene expression include use of SpRYc in complex with guide RNA in a CRISPR-Cas9 system.

Disclosed in the present invention are a sucrose synthase mutant and a method for preparing RD using same. The sucrose synthase mutant is based on the amino acid sequence shown in SEQ ID NO: 2 and contains a mutation in at least one of the following sites: D296Q, R636Q, G514L, H531L, R567T, D569Q, E610N, A642N, E663N, L667T and T742R. By means of site-directed mutagenesis of sucrose synthase, the enzyme activity of the sucrose synthase mutant is improved compared with that of the wild type. Also, some mutants can still maintain a high conversion rate in a wide pH range.

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker positioned on a No.15 chromosome of a sow and related to the lactation ability of the sow. The SNP site of the SNP molecular marker corresponds to the Agt at the 31884611bp position on the No.15 chromosome of an international pig reference genome version 11.1; g mutation; the genotype of the gene is AA, AG or GG. The SNP molecular marker provided by the invention is remarkably related to the sow lactation ability character, the identification of the sow lactation ability character can be realized by identifying the single nucleotide polymorphism of the SNP molecular marker and / or the genotype of the SNP molecular marker, and the dominant allele frequency can be increased generation by generation and the sow lactation ability can be improved by preferably selecting the dominant allele G of the SNP provided by the invention. The pig genetic improvement progress is accelerated, so that the economic benefit of breeding pig breeding is effectively improved.

The present invention belongs to the technical field of genetic engineering. Provided are a single-base editor, a deaminase used therein, and the use thereof. The technical problems to be solved are to identify a naturally occurring cytosine deaminase without sequence preference, construct a base editor, and improve the efficiency and scope of base editing. In order to solve the technical problems above, a cytosine base editor is provided. The cytosine base editor is a fusion protein, wherein the fusion protein is a protein containing a cytidine deaminase, a Cas protein and a uracil-DNA glycosylase inhibitor, and the cytidine deaminase is a protein having an amino acid sequence of positions 28-164 of SEQ ID NO. 2. Further provided is the use of the fusion protein above and a biomaterial related thereto in plant single-base editing. The single-base editor can improve the efficiency of cytosine base editing and accurately mediate the base mutation of a target, and is widely applicable in the cells of maize and even other plants.

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

The invention discloses a dihydrodaidzein reductase, a combinatorial enzyme and an application of the dihydrodaidzein reductase and the combinatorial enzyme in synthesis of S-equol. The dihydrodaidzein reductase is a mutant which takes the dihydrodaidzein reductase AeDHDR as a starting sequence and has a corresponding mutation site, wherein the dihydrodaidzein reductase AeDHDR is derived from AdleCRzia equaliforns, and the amino acid sequence of the dihydrodaidzein reductase AeDHDR is shown as SEQ ID NO: 2. The invention also discloses a coding gene, a carrier and a bioengineering bacterium of the dihydrodaidzein reductase mutant and a method for synthesizing S-equol. The method has the advantages of high enzyme activity and high S-equol yield.

The invention provides application of a BnaXPT gene in regulating and controlling the content of oil and protein in rape seeds. According to the invention, the key gene XPT gene for regulating and controlling the balance of the oil content and the protein content of the rape seeds is excavated on the whole genome level through multi-omics association analysis, and by constructing an XPT overexpression strain and an XPT function-deficient rape variety, it is found that the protein content of the rape seeds can be remarkably improved through overexpression of the gene; by mutating the gene, the oil content of rape seeds can be remarkably increased, a brand new target is provided for improving and optimizing the content and balance of the two nutrient substances of the rape, a new way and means are provided for creating the rape with high oil content or high protein content, and the application prospect is wide.

The invention provides a construction method, system and product of a false positive judgment model of a mutation site obtained by sequencing after enzyme digestion fragmentation and related application. Specifically, the false positive judgment model is constructed on the basis of a plurality of characteristic parameters associated with mutation sites obtained by enzyme digestion fragmentation sequencing; the characteristic parameters comprise scores obtained by performing reverse complementary sequence alignment on the front and back extension 30bp of the mutation site, the sequence proportion of 15 basic groups at the tail end of the mutation site, the Bayesian conditional probability of the extracted mutation site, and the KS test statistic of the extracted mutation site. Furthermore, the invention also provides a method for constructing the model by utilizing the crowd detection rate and the corresponding false positive. According to the method, the data characteristics of the false positive mutation sites are introduced by utilizing digestion fragmentation, so that the reliability of the digestion false positive mutation sites is objectively evaluated under the condition that high depth and large data volume are not needed, and the detection accuracy of the mutation sites can be remarkably improved.