217 results about "Mutant gene" patented technology

The invention discloses a male sterility gene ZmUCE4 and application thereof in creating a male sterility line of corn, and belongs to the field of plant biotechnology breeding. According to the invention, site-directed mutagenesis of the ZmUCE4 gene in wild type corn is carried out through a CRISPR / Cas9 gene editing technology, pollen development abnormality and abortion can be caused, complete male sterility is caused, and it is proved that the ZmUCE4 gene has a regulation and control function on corn male reproductive development. Through offspring screening, a sterile line without transgenic components can be obtained, a stable maize male sterile line is created, and the method has important significance on maize male fertility control and hybrid seed production. The invention also designs a functional molecular marker aiming at the obtained uce4 male sterility mutant gene, and has important application value in maize male sterility line cultivation, sterile hybrid seed production and molecular marker-assisted selection.

The invention discloses a sorghum brown midrib mutant gene bmr34 related to high digestibility and application of the sorghum brown midrib mutant gene bmr34. The haplotype of the mutant gene bmr34 can specifically reduce the lignin content by 2.99% without affecting the biological yield. The invention provides the KASP marker which can accurately locate the bmr34 gene of sorghum and is closely linked with the bmr34 gene of sorghum, so that the breeding efficiency can be greatly improved, the breeding process is accelerated, and the KASP marker has important theoretical and practical significance for improving the quality of sorghum feed.

The invention discloses a rice seed dormancy gene OsPHS1 and application of a protein encoded by the gene OsPHS1, and belongs to the technical field of plant genetic engineering, and the gene and the protein encoded by the gene are used for regulating the dormancy of rice seeds. The nucleotide sequence of the gene is shown as Seq ID No: 1, the CDS sequence is shown as Seq ID No: 2, and the coded protein sequence is shown as Seq ID No: 3. The function of the rice seed dormancy gene OsPHS1 is verified by cloning and identifying the rice seed dormancy gene OsPHS1, and research finds that the dormancy of rice seeds can be remarkably reduced by knocking out or mutating the OsPHS1 gene, and the dormancy of the rice seeds can be improved to a certain extent by overexpressing the gene; the invention plays an important role in developing rice varieties with moderate dormancy.

The application discloses a carbonyl reductase mutant and application thereof in preparation of (R)-8-chloro-6-hydroxyoctanoate compounds. The application relates to a mutant of carbonyl reductase with improved catalytic performance and stereoselectivity after rational molecular modification, a recombinant expression plasmid containing the mutant gene of the enzyme, a recombinant expression transformant, a preparation method of the recombinant enzyme, and application of the recombinant enzyme or the recombinant cell as a catalyst in asymmetric reduction of 8-chloro-6-carbonyloctanoate compounds (I) to prepare (R)-8-chloro-6-hydroxyoctanoate compounds (II), in particular, in an intermediate (R)-8-chloro-6-hydroxyoctanoate methyl ester of thioctic acid. The disclosed carbonyl reductase SsCR mutant has the advantages of high reaction efficiency and high catalytic activity, and exhibits the advantages of high substrate concentration, low catalyst consumption and short reaction time in the process of catalytic preparation of the compound (II).

This invention discloses a laccase mutant, Lcc5-I479T, its expression strain, and its applications. Using laccase from *Coprinopsis cinerea* as the starting enzyme, this invention obtained the mutant gene through molecular docking and positional analysis. After induction expression in engineered bacteria containing the mutant gene, the stable laccase mutant enzyme Lcc5-I479T was obtained. Using ABTS as a substrate, the mutant enzyme Lcc5-I479T showed a stable increase in activity to 8-20 times that of the starting enzyme at 35-50℃. This mutant enzyme has potential application value in the oxidative transformation of aflatoxin.

The invention relates to the technical field of molecular biology, in particular to a multiplex PCR (polymerase chain reaction) molecular marker amplification system for identifying Waxy mutant genes of wheat, a product and application of the multiplex PCR molecular marker amplification system. The multiple PCR molecular marker amplification system developed by the invention has the advantages of stable amplification and good specificity, realizes simultaneous genotype identification of three waxy genes Wx-A1, Wx-B1 and Wx-D1 through one-time PCR, is simple, convenient and rapid in detection process and high in flux, can accurately and efficiently screen out waxy wheat materials containing Waxy mutant genes, and has good application prospects. The waxy wheat breeding molecular marker-assisted selection efficiency and accuracy can be improved, the breeding process is shortened, and technical support can be provided for waxy wheat breeding work.

Compositions and methods for detecting structural variations (SVs) in target genes or for genetic mapping of movable transposable elements are disclosed, the target genes relating to disease pathologies commonly found in large Mendelian genomics projects, and the movable transposable elements relating to genetic diseases, cancer, and aging. The method comprises: (i) contacting a sample containing genomic DNA with a DNA endonuclease for an effective amount of time to cleave the genomic DNA into fragments, the genomic DNA being uncrosslinked; (ii) subjecting the fragments obtained from step (b) to a DNA ligase to obtain circularized DNA; (iii) subjecting the circularized DNA to reverse PCR amplification containing a reverse primer, wherein the reverse primer is designed to match a expected wild-type sequence near a suspected mutant locus in the gene; and (iv) sequencing the amplified products. Exemplary conditions include Bardet-Biedel syndrome; severe upper and lower limb defects; retinitis pigmentosa; syndromic microcephaly; spastic paraplegia; and atypical hemolytic uremic syndrome.

The invention discloses an organ-like drug resistance prediction method and device and a storage medium, and relates to the technical field of bioinformatics, and the method comprises the steps: S1, constructing an initial prediction model; s2, acquiring a training data set; s3, training an optimization prediction model; s4, acquiring related data of a to-be-predicted drug; s5, obtaining a prediction result of the drug resistance of the to-be-predicted drug; according to the method, the drug resistance of the organoid to the drug can be predicted more accurately. By fusing multi-dimensional features of gene mutation, target mutation and drug functional groups, the understanding and learning ability of the model to the drug resistance mechanism is enhanced, and the reliability of drug resistance prediction is improved. The method not only can accurately capture key information of influence of gene and target mutation on drug resistance, but also can highlight unique mutant genes of the organoid on drug resistance by comparing feature differences of the organoid and a common cell line, so that an efficient and accurate calculation framework is provided for drug resistance detection.

The invention discloses a glucose-6-phosphate dehydrogenase mutant as well as an expression strain and application thereof. According to the invention, on the basis of glucose-6-phosphate dehydrogenase from marine microorganisms, after a mutation site is selected by combining Evcouplings design with Fold X scoring, a recombinant plasmid is constructed, and a mutant gene is obtained. After the engineering bacteria containing the mutant plasmids are subjected to induced expression, the glucose-6-phosphate dehydrogenase G6PDH202-Des3 with improved specific enzyme activity is obtained. Under the condition of 37 DEG C, the specific enzyme activity of the mutant is improved to 1.6 times, and the stability is improved to 1.2 times. The mutant has potential application value in an in-vitro diagnostic kit for detecting creatine kinase activity by an enzyme method.

The purpose of the present invention is to provide a novel mutant gene that imparts perpetual flowering properties to roses. Specifically, the present invention pertains to a KSN mutant gene comprising a base sequence (a) or (b): (a) a base sequence represented by SEQ ID NO: 1; and (b) a base sequence that is at least 90% identical in sequence to the base sequence represented by SEQ ID NO: 1, and maintains a transposon comprising the base sequence from position 729 to position 5812 in the base sequence represented by SEQ ID NO: 1.

The invention relates to the technical field of biomedical treatment, in particular to a primer probe combination for distinguishing and relatively quantifying wild type and secondary drug-resistant mutation of EGFR (Epidermal Growth Factor Receptor) genes. The ARMS PCR primer probe combination, the detection reagent, the kit and the detection method which take the EGFR gene as the target are obtained by optimizing the mutation site position and concentration of the primer probe and screening the annealing temperature and enzyme. The primer probe combination, the detection reagent, the kit and the detection method disclosed by the invention are used for ARMS PCR detection of EGFR genes, the amplification inhibition capability on a wild type template is improved, non-specific amplification is reduced to improve specificity, and meanwhile, a good linear relationship can be kept; the primer design process is simplified, and introduction of extra mismatched bases is avoided; a reliable relative quantitative system is established, experimental interference factors are eliminated, and abundance of mutant genes is accurately reflected; the detection period is shortened, the cost is reduced, clinical rapid detection requirements are met, and a more reliable technical means is provided for detection of mutant genes.

The application discloses TaBLM10 The application belongs to the technical field of gene editing and relates to application of a gene in improving resistance of wheat to scab. TaBLM10 The application improves the expression level of the gene by editing the promoter sequence of the gene TaBLM10 , thereby improving the resistance of wheat to scab. The promoter sequence is a sequence as shown in SEQ ID NO. 4, wherein a G / A mutation exists at the 60th base; a sequence as shown in SEQ ID NO. 5, wherein a G / A mutation exists at the 2373th base; and a sequence as shown in SEQ ID NO. 6, wherein a G / A mutation exists at the 341th base. TaBLM10 The application first finds the correlation between the gene and the resistance of wheat to scab, and improves the expression level of the gene by editing the promoter sequence of the wheat TaBLM10 TaBLM10 gene, thereby enhancing the resistance of wheat to scab and laying a foundation for creating new germplasm resistant to scab.

The invention provides a GmVPS35A1 mutant gene and application thereof, the GmVPS35A1 mutant gene is changed into a Gmvps35a1 mutant gene from a Gmvps35a1 gene, the Gmvps35a1 mutant gene is Gmvps35a1-KO1, Gmvps35a1-KO3 or Gmvps35a1-KO4, and the nucleotide sequence of the Gmvps35a1-KO1, the nucleotide sequence of the Gmvps35a1-KO3 and the nucleotide sequence of the Gmvps35a1-KO3 and the nucleotide sequence of the Gmvps35a1-KO4 are shown as SEQ ID NO.2, SEQ ID NO.3 or SEQ ID NO.4 respectively. The invention further provides a preparation method of the GmVPS35A1 mutant The invention discloses an application of GmVPS35A1 protein in increasing the content of soybean protein. The sequence of the GmVPS35A1 protein is shown as SEQ ID NO.5. The invention further discloses a preparation method of the GmVPS35A1 protein. And a theoretical basis is provided for subsequent preparation of high-protein soybeans.

The invention provides an LMNA mutant gene and application thereof, and belongs to the technical field of biomedicine. The nucleotide sequence of the human LMNA mutant gene is as shown in SEQ ID NO. 1. The LMNA mutant gene can be used for screening dilated cardiomyopathy and atrioventricular block and constructing a non-human animal model of the dilated cardiomyopathy and the atrioventricular block, and is beneficial to exploring pathophysiological processes of the dilated cardiomyopathy and the atrioventricular block and exploring therapeutic targets and drugs of the dilated cardiomyopathy and the atrioventricular block.

The present invention relates to a lettuce plant that is resistant to Fusarium wilt, more specifically to a lettuce plant that comprises a mutated gene that confers resistance to F. oxysporum in lettuce. Furthermore the present invention relates a resistance gene and a method for obtaining a lettuce plant that is resistant to Fusarium wilt, wherein the method comprises the step of mutating a gene.

The invention discloses an acyl-coenzyme A oxidase mutant as well as an expression strain and application thereof. On the basis of acyl coenzyme A oxidase derived from marine microorganisms, mutation sites are designed through Saprot and scored, and mutation is introduced through overlapping extension PCR (polymerase chain reaction), so that a mutant gene is obtained. After the engineering bacteria containing the mutant plasmids are subjected to induced expression, the acyl-coenzyme A oxidase with improved stability and specific enzyme activity is obtained. Under the condition of 37 DEG C, the stability of the mutant is improved by 1.3 times. When palmitoyl coenzyme A is used as a substrate, the specific enzyme activity of the mutant is improved to 1.5 times. The mutant has application value in an in-vitro diagnostic kit for detecting free fatty acid by an enzyme method.

The present application relates to a CRISPR-mediated immortalized sheep fetal skin fibroblast cell line in the field of variation or genetic engineering and application thereof.The construction of the CRISPR-mediated immortalized sheep fetal skin fibroblast cell line of the present application comprises the following steps: step one, constructing a Cas9 site-directed knockout plasmid containing an FGF5 gene target sequence; step two, constructing a homologous repair plasmid Donor-2 kb HA-SV40LT containing an FGF5 gene homologous arm; step three, constructing a donor DNA containing an SV40LT protein expression frame; and step four, preparing a sheep fetal fibroblast cell line stably expressing SV40LT protein.The immortalized SFF-FGF5 / SV40LT monoclonal cell strain created by the present application can become an ideal tool cell line for genome editing researches such as sheep CRISPR library screening, multiple gene editing and mutation gene function exploration.

The invention relates to a seawater oryzias medaka stable expression cas9 cell line, a construction method and application, and belongs to the technical field of gene editing, the cell line takes seawater oryzias medaka cells as a main body, and a cas9-Blasticidin gene is inserted into a seawater oryzias medaka cell genome. The construction method of the cell line comprises the following steps: infecting a testis cell line of oryzias melastigma by lentivirus to realize stable insertion of a cas9 gene in an oryzias melastigma genome, and screening a monoclonal cell line to obviously improve the expression quantity of cas9 in the cell line. The invention also provides a method for constructing the oryzias melastigma cell line by utilizing the cell line to edit the mstnb gene of the oryzias melastigma cell, and the method for constructing the oryzias melastigma cell line. The mstnb gene is successfully mutated by transfecting sgRNAs in the cell line.

This invention discloses a quantitative genomic RNA standard for the BCR-ABL1 fusion gene and its preparation method. The method involves extracting genomic RNA containing two mutant forms of the BCR-ABL1P210 fusion gene, b2a2 and b3a2. RNA storage buffer and quantitative standard solutions are prepared, and corresponding primers and probes are designed. A one-step reverse transcription digital PCR method is used, with the BCR-ABL1P210 fusion mutant genomic RNA as a template for PCR amplification. Fluorescence signals are collected to detect the expression of the BCR-ABL1P210 fusion genes b2a2 and b3a2, thereby obtaining the copy number content of the BCR-ABL1P210 fusion genes b2a2 and b3a2 and the abundance of the mutant gene in ABL-WT as quantitative values.

The invention discloses application of a transcription extension complex subunit ZmELP2 in regulation and control of corn grain development and plant height, and belongs to the technical field of plant genetic engineering and corn molecular breeding. The corn mutant smk251 with stable inheritance is obtained through EMS mutagenesis treatment, and compared with a wild type, the corn mutant smk251 has the advantages that the grain size is obviously reduced, and the plant height is reduced. The mutant provides a source for digging genes for regulating and controlling corn kernel and plant height. Based on a corn mutant smk251, a mutant gene ZmELP2 gene is further obtained through map-based cloning technology positioning, the function of the ZmELP2 gene is verified through overexpression and refilling tests, and it is determined that the mutant smk251 is really caused by ZmELP2 gene mutation. The ZmELP2 gene provided by the invention can be used for providing a new gene resource for cultivating high-yield corn varieties and lodging-resistant corn varieties, and has important production and application values.

The present application belongs to the technical field of genetic engineering, and relates to a CrtI enzyme mutant, a recombinant plasmid, a recombinant strain and application thereof. The CrtI enzyme mutant provided by the present application is obtained by random mutation of a wild-type CrtI gene through error-prone PCR, and the amino acid sequence thereof is shown as SEQ ID NO. 1. The obtained mutant has multiple site variations in the amino acid sequence, thereby significantly improving the catalytic efficiency and substrate affinity of the enzyme. The present application further constructs a recombinant expression vector containing the CrtI enzyme mutant gene, and transforms the recombinant expression vector into a red Phaffia yeast competent cell, and obtains a high-yield astaxanthin recombinant strain through resistance screening and pigment phenotype screening. Compared with the wild-type CrtI enzyme, the CrtI enzyme mutant of the present application significantly improves the conversion rate of beta-carotene to astaxanthin in the red Phaffia yeast.

The invention discloses application of a rice malic acid synthase gene in improving the nutritional quality of seeds. The invention reports that the OsIPMS1 and OsIPMS2 genes can regulate and control the nutritional quality in polished rice grains for the first time in rice, and tests show that the mutation of the OsIPMS1 and OsIPMS2 genes improves the content of protein and amino acid in the polished rice grains. It is proved that the OsIPMS1 and OsIPMS2 genes regulate and control the nutritional quality of rice seeds, and the genes are beneficial to cultivation of rice varieties with high protein and amino acid.

The present invention provides an anti-cancer vaccine comprising: (i) at least one peptide comprising the amino acid sequence of a neoantigen encoded by a mutant homologous recombination (HR) DNA repair gene selected from the group: BRCA1, BRCA2, PALB2, CDK12, RAD51B, RAD51C and RAD51D, wherein the mutant gene comprises a reversion mutation; and / or (ii) at least one polynucleotide encoding the at least one peptide of (i). Also provided are engineered T cells that recognise said neoantigen. Related methods and medical uses of the vaccine and / or engineered T cell are provided, including for the treatment of cancers, such as homologous recombination (HR) deficient cancers that acquire PARP inhibitor resistance or platinum resistance by development of reversion mutations in an HR DNA repair gene selected from the group: BRCA1, BRCA2, PALB2, CDK12, RAD51B, RAD51C and RAD51D.

The invention belongs to the field of crop heredity, and relates to application of a polyphenol oxidase gene. The invention discloses an InDel marker for rice brown glume traits, a regulatory gene and application of the InDel marker and the regulatory gene. Based on a brown glume mutant material in the background of Hunan early indica 42, genetic analysis shows that the mutation is dominant mutation, single-gene control and gene positioning find that the mutant gene is a polyphenol oxidase encoding gene (MSULocus is LOCOs04g53300), and the exon region of the mutant gene is deleted by 6bp (GCGACT), so that the material shows brown glume; the gene is knocked out from a brown glume mutant, and the glume color becomes normal; the method has the advantages that the GCGACTs are used as the raw materials, the GCGACTs are accurately edited in the wild type materials, the corresponding 6bp deletion (GCGACT) materials are obtained, the glumes of the GCGACTs are brown, and new gene resources and methods are provided for quickly and directionally cultivating novel rice materials and rice materials suitable for mechanized seed production of hybrid rice.

This invention discloses a bifunctional flavin adenine dinucleotide synthase mutant and its application in FAD synthesis. The mutant disclosed in this invention is RFK / FADS. C18R or RFK / FADS C18R‑R46H ;RFK / FADS C18R The amino acid sequence is shown in SEQ ID NO.15, RFK / FADS C18R‑R46H The amino acid sequence is shown in SEQ ID NO. 16. Genetically engineered bacteria carrying the bifunctional flavin adenine dinucleotide synthase mutant gene plasmid produced flavin adenine dinucleotide using glucose as a substrate through a simple fermentation process under aerobic conditions in a shake flask. The FAD yields of strains XF02 and XF03 containing the mutant gene were 79.31 mg / L and 167.56 mg / L, respectively, representing increases of 58.62% and 235.12% compared to strain XF01 containing the non-mutated gene. By constructing microbial "cell factories" through metabolic engineering strategies, modified microorganisms can utilize inexpensive raw materials to generate desired natural products within their cells. This advancement not only reduces the production cost of FAD but also improves the environmental sustainability of its production process.

The application discloses a method for constructing a myopathy animal model, the constructed model and application of the model. The application also provides application of a reagent for knocking out or mutating a PRR14 gene or inhibiting expression of the gene in preparation of a myopathy animal model product. Specifically, compared with normal mice, total muscle mass, grip level and muscle endurance of the mice with muscle tissue-specific knockout of the PRR14 gene are all significantly reduced, the animal model provided by the application has typical characteristics of myopathy, can well simulate occurrence and development of myopathy in vivo, is an ideal animal model for basic and clinical application research of myopathy, and can be well applied to screening of drugs for treating myopathy.

The present invention relates to a modified U1 snRNA configured to rescue exon skipping in a mutant ABCA4 gene. In one aspect of the invention, there is provided a modified U1 small nuclear RNA (snRNA) configured to retain exon 40 in a mature mRNA transcript of a mutant ABCA4 gene comprising a mutation that induces exon 40 skipping wherein the mutation is located between 3 base pairs upstream and 8 base pairs downstream of a donor splice site of exon 40, wherein the modification comprises replacing a portion of the single-stranded nucleotide sequence of the 5'region of the wild-type U1 snRNA with a single-stranded binding nucleotide sequence capable of hybridizing to a target sequence present on the precursor mRNA transcript of the mutant ABCA4 gene, and wherein the target sequence is located in a region between 13 base pairs upstream and 15 base pairs downstream of the donor splice site of exon 40. In another aspect, there is provided a nucleic acid construct comprising a polynucleotide sequence encoding a modified U1 snRNA as described herein.

This invention relates to the field of agricultural technology, specifically to a mutant rice OsDSP1 gene and its applications. Compared to the wild-type rice OsDSP1 gene, the mutant OsDSP1 gene has a T base inserted after position 115 or a G base deleted at position 116. The mutant rice OsDSP1 gene provided by this invention can improve rice resistance to brown planthoppers, while also increasing grain length and thousand-grain weight, effectively increasing rice yield. This invention provides new gene resources and improvement targets for the breeding of insect-resistant rice varieties, and has significant application value for insect-resistant and yield-increasing plant breeding.

The application discloses a CRP mutant gene and application thereof, and belongs to the technical field of genetic engineering. The amino acid sequence of the CRP mutant gene is shown in SEQ ID NO. 7 or SEQ ID NO. 8 or SEQ ID NO. 9, the CRP mutant gene can significantly eliminate the 'glucose inhibition' effect, activate glycolysis and TCA cycle in a basic medium with glucose as a carbon source, strengthen the utilization and biosynthesis of glucose, and promote the growth of a strain, and the CRP mutant gene with the amino acid sequence shown in SEQ ID NO. 7, 8 or 9 also has the effect of improving the expression amount of vanillin or naringenin of the strain.

The application discloses a male sterility gene Zmuce4 and application thereof in creating a maize male sterile line, and belongs to the field of plant biotechnological breeding. The application proves that the gene has a regulating function on maize male reproductive development by causing abnormal pollen development and abortion and leading to complete male sterility through site-directed mutation of the gene in wild-type maize by CRISPR / Cas9 gene editing technology. Zmuce4 Through offspring screening, a sterile line without a transgenic component can be obtained, and a stable maize male sterile line is created, which has important significance for maize male fertility control and hybrid seed production. uce4 The application also designs a functional molecular marker for the obtained male sterility mutant gene, which has important application value in maize male sterile line breeding, sterile hybrid seed production and molecular marker assisted selection.