322 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 relates to the technical field of gene engineering, in particular to an L-rhamnose isomerase mutant, a recombinant plasmid, a mutant strain and application. The amino acid sequence of the mutant is as shown in SEQ ID NO.4, and the nucleotide sequence of the coding gene is as shown in SEQ ID NO.3. A specific mutant gene sequence is designed, a recombinant plasmid pET28a (+)-L-Rhi is successfully constructed, the gene sequence is connected between enzyme cutting sites BamH I and Hind III of an escherichia coli expression vector pET-28a (+) to construct a mutant expression vector (the recombinant plasmid pET-28a (+)-L-Rhi), the mutant expression vector is transformed into an escherichia coli BL21 competent cell, a mutant strain is obtained, and the recombinant plasmid pET-28a (+)-L-Rhi is obtained. The obtained mutant strain can overexpress L-rhamnose isomerase, the conversion rate of D-allose is remarkably improved, the thermal stability of the enzyme is remarkably improved through the design of mutation sites, and the half-life period is effectively prolonged.

The invention relates to the technical field of gene engineering, in particular to an L-rhamnose isomerase mutant, a recombinant plasmid, a mutant strain and application. The amino acid sequence of the mutant is as shown in SEQ ID NO. 2, and the nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1. A specific mutant gene sequence is designed, a recombinant plasmid pET28a (+)-L-Rhi is successfully constructed, the gene sequence is connected between enzyme cutting sites BamH I and Hind III of an escherichia coli expression vector pET-28a (+) to construct a mutant expression vector (the recombinant plasmid pET-28a (+)-L-Rhi), the mutant expression vector is transformed into an escherichia coli BL21 competent cell, a mutant strain is obtained, and the recombinant plasmid pET-28a (+)-L-Rhi is obtained. The obtained mutant strain can overexpress L-rhamnose isomerase, the conversion rate of D-allose is remarkably improved, the thermal stability of the enzyme is remarkably improved through the design of mutation sites, and the half-life period is effectively prolonged.

The application discloses a kind of fungal laccase mutant Lcc5-G and its expression strain and application.The application is based on the fungal laccase from Coprinopsis cinerea heterologous expression since, by site-directed mutagenesis, obtain mutant gene.After fungal induction expression containing mutant plasmid, obtain laccase mutant enzyme Lcc5-G with improved specific activity, stability and AFB1 removal efficiency.When ABTS is used as substrate, the specific activity of the mutant is improved by 3 times, and the stability of the mutant enzyme is improved to 5 times of the original enzyme at 45 DEG C and pH 7.The mutant has potential application value in oxidizing aflatoxin.

Using chemical genetic screening, we discovered a small molecule Cellulosin (aka endosidin20 or ES20) that causes cell swollen and inhibits plant growth, but does not disrupt global vesicle trafficking. By doing mutant screening, we obtained multiple alleles of Arabidopsis thaliana that are resistant to Cellulosin inhibition in growth. Those mutated amino acid residues are conserved across plant species. Cellulosin targets a group of cellulose synthases (CesAs) of Arabidopsis thaliana by binding to a conserved domain essential for the catalytic activity of CesA. Cellulosin may target and inhibit all subtypes of CesAs in plants. The present invention relates to Cellulosin, a cellulose synthase inhibitor, its analogs or derivatives as a broad-spectrum herbicide. The mutated genes, their protein products and a cell or a plant having those mutated genes or expressing those protein products are within the scope of this disclosure.

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.

In traditional plant breeding approaches, chemical mutagenesis may be utilized to introduce nucleotide substitutions at random in the genome of a plant, i.e. without possibilities to control the sites of nucleotide changes. Because of genome complexities, the statistical probability is extremely little when it comes to finding a predetermined nucleotide substitution. The present invention, however, demonstrates how a novel, alternative use of digital polymerase chain reaction (dPCR), preferably droplet dPCR (ddPCR), is developed to exploit finding of specific nucleotide substitutions in mutated genes. The entire platform comprises a screening method with a library of mutagenized organisms, digital PCR-based systems and a set-up to propagate and analyze identified, mutated organisms.

The invention relates to a gene ZmILR1 for controlling the size of a corn kernel as well as a molecular identification method and application of the gene ZmILR1, and belongs to the technical field of plant genetic breeding and functional gene research. The ZmILR1 gene encodes an IAA-amino acid hydrolase, mutation of the IAA-amino acid hydrolase causes corn kernels to become smaller, and overexpression of the gene can create a corn material with increased kernel length, kernel width and hundred-kernel weight. A functional molecular marker A1A4 is developed according to the sequence difference of the mutant gene and a wild type gene at the DNA level, and can be used for rapid identification and screening of small-grain corn materials. The invention provides a theoretical basis and an application tool for analyzing a molecular mechanism of the corn grain size and developing molecular marker-assisted breeding and hybrid seed breeding, and provides important gene resources and technical support for high-yield and high-quality molecular breeding of corn.

The invention discloses a corn ALS mutant gene and application of the corn ALS mutant gene in herbicide resistance. The invention provides a method for improving the herbicide resistance of corn, which comprises the following steps: mutating 493th nucleotides of a ZmALS1 gene and a ZmALS2 gene in a corn genome from C to G so as to improve the herbicide resistance of the corn, the nucleotide sequence of the ZmALS1 gene is shown in SEQ ID No. 1; the nucleotide sequence of the ZmALS2 gene is shown in SEQ ID No. 2. Specific sites in corn ZmALS1 and ZmALS2 serve as sgRNA recognition sites on the basis of the principle of a base editing technology, a single base replacement editing vector capable of simultaneously targeting the ZmALS1 and ZmALS2 is designed, the gene editing vector is stably transformed through agrobacterium to obtain a transgenic plant, and the resistance to herbicides is remarkably improved.

The invention belongs to the field of biotechnology and environmental governance, and particularly relates to a manganese peroxidase mutant modified through semi-rational design and application of the manganese peroxidase mutant to straw degradation. On the basis of Irpex lacteus MnP, a core site G189 is screened out through structural analysis and alanine scanning, a G189A single mutant (glycine-alanine) is constructed, the G189A single mutant is free of Mn < 2 + > dependence, the enzyme activity reaches 15.2 U / mL (about 130% higher than that of a wild type), and the catalytic efficiency is improved by 2.1 times. Meanwhile, the mutant gene is introduced into bacillus subtilis to realize efficient extracellular secretion, the straw lignocellulose depolymerization can be effectively promoted, the application value in the fields of agricultural straw recycling and the like is remarkable, and the wide market application prospect is achieved.

The invention discloses yarrowia lipolytica with high yield of beta-carotene as well as a construction method and application of the yarrowia lipolytica. According to the yarrowia lipolytica engineering bacterium, geranyl diphosphate synthase gene xdGPS, phytoene dehydrogenase gene CarB, phytoene cyclization / phytoene synthesis bifunctional enzyme mutant gene GarRPY27R, acetyl-coenzyme A synthetase mutant gene ACSL641P and pyruvate ferredoxin oxidoreductase gene nifJ are integrated and expressed on a chromosome, and the yarrowia lipolytica engineering bacterium is obtained. Meanwhile, a 3-hydroxy-3-methylglutaryl CoA reductase gene HMGR (3-hydroxy-3-methylglutaryl CoA reductase) derived from the yarrowia lipolytica strain is subjected to overexpression; the yield of the beta-carotene is increased to 13.186 g / L, and the production efficiency is 0.14 g / L / h.

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 bactrocera dorsalis point mutation gene editing method based on CRISPR-Cas9 and homologous directional repair, and relates to the technical field of gene editing. The method comprises the following steps: designing high-specificity sgRNA for a PAM sequence adjacent region of a target gene; extending the length of the homologous arm to 100bp, and designing a single-stranded DNA (deoxyribonucleic acid) containing a mutation site as a homologous repair template; after a mixture containing the high-specificity sgRNA, the homologous repair template and Cas9 protein is injected into a bactrocera dorsalis fertilized egg in a microinjection mode, incubation and incubation are carried out, and G0-generation imagoes are obtained through culture; hybridizing the G0-generation imago with wild bactrocera dorsalis, and screening to obtain a G1-generation hybrid mutant; and carrying out selfing by using the G1-generation heterozygous mutant, and screening to obtain a G2-generation homozygous mutant. By adopting the method, the mutation efficiency and embryo survival rate of bactrocera dorsalis can be effectively improved.

The invention is applicable to the technical field of molecular biology, and provides a mutant gene, a mutant, a transformant and a genetic transformation plant of tobacco deoxycarboxy-putrescine lysine synthase and application of the mutant gene, the mutant, the transformant and the genetic transformation plant of the tobacco deoxycarboxy-putrescine lysine synthase of the tobacco deoxycarboxy-putrescine lysine synthase of the tobacco deoxycarboxy-putrescine lysine synthase. The nucleotide sequence of the mutant gene is shown as SEQ ID NO.2 in a sequence table. After the gene is mutated, a mutant strain shows the phenotype of white veins, and various nutrients are greatly changed. According to the mutant gene of the tobacco deoxycarboxamide lysine synthase provided by the invention, by detecting the mutant and comparing the potassium content in tobacco leaves, the result shows that the mutant gene of the tobacco deoxycarboxamide lysine synthase can improve the potassium content in the tobacco leaves. Therefore, a better choice is provided for genetic improvement of new plant varieties, and the gene has important application value.

Provided are a rice mutant gene and the use thereof in regulating the plant height of rice. The rice mutant gene at least has an allele mutation, and the allele mutation is the mutation into Asn from Asp at the 377th site in an amino acid sequence shown as SEQ ID No. 26. The mutant gene can effectively regulate and control the rice plant height, and has good application potential in plant architecture breeding of rice varieties.

The present invention provides a mutant gene, detection and application that causes 3-methylglutaconic aciduria type VII. The mutant gene that causes 3-methylglutaconic aciduria type VII includes a compound heterozygous mutation at the CLPB:NM_030813.6:exon8:c.1016T>G:p.L339R site and the exon1:c.130delG:p.E44Sfs*5 site. The mutant gene can effectively distinguish patients with 3-methylglutaconic aciduria type VII from the normal population. Therefore, the pathogenic gene mutation of the present invention can be used as a biomarker for diagnosing 3-methylglutaconic aciduria type VII. The present invention can be used for screening or diagnosing the genetic diagnosis of 3-methylglutaconic aciduria type VII by detecting whether the subject carries the above-mentioned mutation. The detection kit provided by the present invention can be used to quickly and effectively predict or diagnose 3-methylglutaconic aciduria type VII.

The invention discloses a mutant gene OsNRAMP5 capable of reducing cadmium content in rice grains and application of the mutant gene OsNRAMP5. The gene has a genome nucleotide sequence as shown in SEQ ID No.1, and the 8875149th nucleotide on the No.7 chromosome of the wild type of fragrant rice, namely the 10th exon splicing site of the LOCOs07g15370 (OsNRAMP5) gene, is mutated from A to T, so that the gene is subjected to variable splicing in the transcription process. The invention also discloses a protein coded by the nucleotide sequence and used for regulating and controlling low accumulation of cadmium in fragrant rice grains, and the protein has an amino acid sequence as shown in SEQ ID No.3. The OsNRAMP5 gene for regulating and controlling the accumulation of the harmful heavy metal element cadmium is cloned in the fragrant rice by utilizing a forward genetics technology, and the mutant gene not only can improve the content of fragrant substances in the fragrant rice, but also can greatly reduce the content of cadmium in the rice, so that the quality safety of the fragrant rice is ensured.

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 invention provides an application and a method of a pineapple zeaxanthin epoxidase AcoZEP1 gene, and belongs to the technical field of biology.The application is that the AcoZEP1 gene is inactivated or the activity of the AcoZEP1 gene is reduced, so that the content of ABA in pineapple leaves and the activity of ZEP in the pineapple leaves are reduced, and therefore pineapple leaf thorns are reduced; according to the method, a pineapple mutant for inactivating or reducing the activity of an AcoZEP1 gene is planted so as to reduce pineapple leaf thorns. By mutating the AcoZEP1 gene, the density of pineapple leaf thorns can be remarkably reduced, and the thorn-free pineapple germplasm can be cultivated by using molecular breeding technologies such as transgenosis and gene editing in the later period, so that data support and material support are provided for solving the problems of long time consumption, time consumption and labor consumption of cross breeding of the thorn-free pineapple variety in the pineapple industry; and the time required for thorn-free breeding of the pineapples is obviously shortened.

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 discloses a beta-farnesene synthase mutant, which is characterized in that the amino acid sequence of the beta-farnesene synthase mutant has the difference of one or more amino acid residues selected from A26T, A169E, Y251H, V323S, G325A and A438T relative to the amino acid sequence of wild type beta-farnesene synthase derived from walnuts. The beta-farnesene synthase mutant gene and the encoding protein thereof provided by the invention are effectively expressed in saccharomyces cerevisiae. Under the same condition, the walnut-derived beta-farnesene synthase mutant provided by the invention not only has higher activity compared with the beta-farnesene synthase derived from artemisia annua, German chamomile, peppermint and fragrant citrus, but also has significantly improved enzyme activity compared with the wild walnut beta-farnesene synthase; and the saccharomyces cerevisiae can have higher farnesene yield.

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.