1352 results about "Mutant gene" patented technology

The present invention provides a method for identifying one or more low abundance sequences differing by one or more single-base changes, insertions, or deletions, from a high abundance sequence in a plurality of target nucleotide sequences. The high abundance wild-type sequence is selectively removed using high fidelity polymerase chain reaction analog conversion, facilitated by optimal buffer conditions, to create a restriction endonuclease site in the high abundance wild-type gene, but not in the low abundance mutant gene. This allows for digestion of the high abundance DNA. Subsequently the low abundant mutant DNA is amplified and detected by the ligase detection reaction assay. The present invention also relates to a kit for carrying out this procedure.

The present invention intends to find out a gene participating in addition of mannose phosphate to a sugar chain of a glycoprotein originating in a yeast belonging to the genus Pichia and provide a means of controlling the same. The present invention also intends to provide a process for producing a protein with reduction of an acidic sugar chain by using the thus controlled yeast strain belonging to the genus Pichia. Namely, the present invention includes a protein participating in the addition of mannose phosphate to a sugar chain of a glycoprotein; a gene encoding this protein; a mutant of this gene; a vector carrying the mutant gene; a yeast strain belonging to the genus Pichia having been transformed by this vector; a process for producing a protein with reduction of an acidic sugar chain by using the transformed yeast strain; and a glycoprotein thus produced.

Seeds, plants and oils are provided having high oleic acid; low linoleic acid; and low linoleic acid plus linolenic acid; and advantageous functional or nutritional properties. Plants are disclosed that contain a mutation in a delta-12 or delta-15 fatty acid desaturase gene. Preferred plants are rapeseed and sunflower plants. Plants carrying such mutant genes have altered fatty acid composition in seeds. In one embodiment, a plant contains a mutation in a region having the conserved motif His-Xaa-Xaa-Xaa-His, found in delta-12 and delta-15 fatty acid desaturases. A preferred motif has the sequence His-Glu-Cys-Gly-His. A preferred mutation in this motif has the amino acid sequence His-Lys-Cys-Gly-His. Nucleic acid fragments are disclosed that comprise a mutant delta-12 or delta-15 fatty acid desaturase gene sequence.

Plants are disclosed that contain a mutation in a delta-12 or delta-15 fatty acid desaturase gene. Preferred plants are rapeseed and sunflower plants. Plants carrying such mutant genes have altered fatty acid composition in seeds. In one embodiment, a plant contains a mutation in a region having the conserved motif His-Xaa-Xaa-His, found in delta-12 and delta-15 fatty acid desaturases. A preferred motif has the sequence His-Glu-Cys-Gly-His. A preferred mutation in this motif has the amino acid sequence His-Lys-Cys-Gly-His. Nucleic acid fragments are disclosed that comprise a sequence of at least 20 nucleotides from a mutant delta-12 or delta-15 fatty acid desaturase gene.

The invention provides an enrichment and analysis method for circulating tumor cells. The method mainly comprises the following steps: subjecting a biological fluid sample to red blood cell lysis treatment; adding immune beads specifically bonding with white cells, red cells and/or platelets for incubation; and filtering cell suspension by using a filter membrane and enriching circulating tumor cells. According to the invention, through integration and optimization of CTCs enrichment methods such as red blood cell lysis treatment, an immune bead method and membrane filtration, the enrichment effect of the circulating tumor cells is greatly improved; with the method, the removal rate of leukocytes is greater than 99.9%, and the enrichment rate of tumor cells is greater than 80%. Meanwhile, the invention also provides a kit using the enrichment method for the circulating tumor cells and discloses a device for enrichment of the circulating tumor cells. The invention also provides an analysis method for the enriched circulating tumor cells. With the analysis method, analysis of dynamic changes like the amount of tumor cells, gene mutation and gene expression profiles can be realized.

The invention provides primers, probes, kit and method for detecting human EGFR (epidermal growth factor receptor) gene mutations. The method comprises the following steps of: (1) providing the primers and the probes; (2) processing detected samples and extracting DNA (deoxyribonucleic acid); (3) carrying out fluorescent quantitation on components of a PCR (polymerase chain reaction) system; (4) amplifying target sequences of an EGFR gene mutation to be detected; and (5) distinguishing wild type genes and mutant type genes by utilizing ARMS (amplification refractory mutation system) primers and judging results via the fluorescence intensity of an FAM and a JOE. The primers, probes and method provided by the invention have the beneficial effects that 29 kinds of mutations on the EGFR genes can be simultaneously detected, so that the sensitivity is high, the specificity is strong and the detection speed is high; and the whole detection process only takes 60 minutes.

The invention discloses a primer and probe for detecting a human epidermal growth factor receptor (EGFR) gene and/or a K-ras gene, a kit containing the primer and the probe and a device for detecting genetic mutation on the basis of a digital PCR platform.The method for detecting the genetic mutation by means of the primer and the probe comprises the steps that the prime and the probe are provided; DNA of a sample to be detected is extracted; a fluorescent PCR reaction system capable of amplifying a mutant gene sequence is prepared; a target probe and an internal reference probe are utilized to be hybridized with amplified products respectively, and fluorescent signals of corresponding fluorescent groups are detected; existence of the genetic mutation is judged and/or the mutation rate is calculated according to the strength and proportion of the fluorescent signals of the target probe and the internal reference probe.According to the method for detecting the genetic mutation, the needed primers and probes are small in number, the optimization procedure is simple, related mutation of EGFR and/or K-ras gene can be qualitatively or quantitatively detected, and the detection sensitivity is high; a DNA sample with low initial amount can also be detected stably.

The invention provides an STAG2 gene mutant sequence and a detection method thereof as well as use of STAG2 gene mutation in detecting bladder cancer and belongs to the technical field of the molecular biology gene detection. The detection method comprises the following steps: (1) extracting a tumor tissue DNA and a peripheral blood DNA from a tumor tissue and the peripheral blood of a bladder cancer patient; (2) performing whole genome sequencing and whole exon sequencing on the two DNAs extracted in the step (1), respectively; (3) performing whole exome sequence alignment and somatic mutation detection on the sequencing results of the step (2), thereby obtaining a mutant gene; (4) verifying the somatic replacement, insertion and deletion of the mutant gene obtained in the step (3) by use of Sanger sequencing; and (5) identifying the mutant gene as the remarkable mutant gene. The detection method has the advantage that the mutation of the STAG2 is one of unfavorable indexes of the bladder cancer and also is capable of serving as a new acting target for treating the bladder cancer.

The invention discloses an SNP (Single Nucleotide Polymorphism) locus related to a maize plant high character, which is the nucleotide 5248 from the 5' tail end in SEQ ID NO:1, the nucleotide 5248 from the 5' tail end in SEQ ID NO:1 is positioned in the 5th exon of a maize dwarf mutant gene br2 in a maize genome, and the nucleotide 5248 is T. Molecular markers of the invention can be used for maize breeding in different genetic backgrounds, can be used for the genetic improvement of maize; and the method and the marks have wide application prospects in the field of maize breeding.

The present invention relates to a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of a mutant gene, comprising a complementary RNA strand having a complementary region that is substantially complementary to a portion of the mutant gene, and which is partially complementary to the corresponding wild-type gene. The invention further relates to a pharmaceutical composition comprising the dsRNA and a pharmaceutically acceptable carrier. The pharmaceutical compositions are useful for inhibiting the expression of a target mutant gene, as well as for treating diseases caused by expression of the target gene. The invention also relates to methods for inhibiting the expression of a target mutant gene, as well as methods for treating diseases caused by the expression of the target gene.

The invention relates to a disease risk predication method and system. The method comprises the following steps of: S1, receiving input gene sequencing result information; analyzing the input gene sequencing result information to obtain all mutant gene information; S2, looking up disease gene mutant information of corresponding gene sites from a precision medical knowledge base according to the gene sites corresponding to the mutant gene information, so as to obtain at least one piece of disease gene mutant information; S3, matching each piece of the disease gene mutant information with all the disease gene mutant information to obtain the similarity of each piece of the mutant gene information, and all the disease gene mutant information; and S4, obtaining a risk predication table aiming at the gene sequencing result information according to the similarity of each piece of the mutant gene information, and all the disease gene mutant information. By analyzing an association relation of susceptibility genes, mutation sites, mutation types and diseases, disease risk predication is carried out on healthy people and general disease risk grade predication is provided; disease risks in the future are reduced.

The present invention discloses a method of detecting a wild-type or mutant B-RAF gene in a body fluid sample from a subject. Also disclosed are methods of using B-RAF as a biomarker for detecting cancer, predicting the outcome of cancer, and monitoring the treatment of cancer or the status of cancer. Furthermore, the invention discloses methods and compositions for detecting a mutant gene with a peptide nucleic acid clamp capable of hybridizing to a wild-type gene and a locked nucleic acid probe capable of hybridizing to a mutant of the gene.

The invention discloses a glucose oxidase mutant gene, expression and application thereof. In the invention, 272 basic groups are changed through codon optimization and GC content change and the content of GC is reduced to 48.44% from 55.54% so as to obtain the glucose oxidase mutant gene, wherein the basic group is represented by SEQ ID NO.2. The glucose oxidase mutant gene is transferred into pichia yeast to express; the experimental result shows that the secretory expression level of the glucose oxidase mutant gene in the pichia yeast is significantly improved by comparing with the same before mutation; compared with partial research at home and abroad, the final secretory expression of the glucose oxidase mutant gene achieves high expression level, thereby building the foundation for further expansion of industrial production. The determination of enzymatic properties of the glucose oxidase mutant gene shows that the recombinant glucose oxidase protein expressed by the glucose oxidase mutant gene has good thermal stability and high enzyme activity.

The invention discloses an omega-aminotransferase mutant gene and an application thereof, and belongs to the field of the molecular biological technology. The nucleotide sequence of the omega-aminotransferase mutant gene is shown as SEQ ID NO.1. The omega-aminotransferase amino acid sequence embodied by the code of the omega-aminotransferase mutant gene is shown as SEQ ID NO.2. The omega-aminotransferase mutant gene is firstly provided, and the omega- aminotransferase corresponding to the code of the omega-aminotransferase mutant gene has a high enzyme amount, so that the biological catalysis and conversion efficiency is improved in industrial production. The invention further provides the application of the omega-aminotransferase embodied by the code of the omega-aminotransferase mutant gene in preparation of trifluoro-propylamine. The trifluoro-propylamine is added to a buffer solution to serve as the substrate for the reaction, and then products of trifluoromethyl amine compounds are collected from the reaction liquid. The method for preparing the trifluoro-propylamine has the obvious advantages that the reaction conditions are mild, no pollution is caused, and the process is simple.

The invention discloses a bacterial laccase mutant protein, which is characterized in that the mutant protein amino acid sequence is obtained by deletion mutation of the 323rd glycine residue to the 332rd glycine residue in the bacterial laccase amino acid sequence shown as SEQ ID No.1. Through a genetic engineering reconstruction method, a stability improved bacterial laccase protein coding gene, its expression plasmid and engineered bacteria can be obtained, and after large-scale fermentation and induced expression of the engineered bacteria, the stability improved bacterial laccase protein can be obtained. According to the invention, the marine uncultured microorganism source bacterial laccase Lac15 is taken as the foundation, and by means of genetic engineering reconstruction, mutant gene can be obtained. At the same time, a recombinant escherichia coli is employed to conduct high-density culture for high-efficiency expression of the bacterial laccase mutant protein. According to the invention, the stability and yield of the bacterial laccase are greatly improved.

The invention discloses a Bacillus subtilis adenylosuccinate synthetase mutant gene purA and applications thereof. The sequence of the Bacillus subtilis adenylosuccinate synthetase mutant gene purA is shown in SEQ ID No.1. Engineering bacteria containing the Bacillus subtilis adenylosuccinate synthetase mutant gene purA (C725T) and built according to the invention are good in bio-safety and clear in genetic background, and the contained mutant gene purA (C725T) can significantly improve the riboflavin synthesis capacity of Bacillus subtilis, and under the condition of fermentation in a shaking flask, the level of riboflavin accumulation is increased by over 40%.

The invention provides a screening method for a tumor new antigen, and the method comprises: A, obtaining polypeptide sequences encoded corresponding to mutant genes of a tumor tissue cell; B, performing predicting on the affinity of each polypeptide sequence with a major histocompatibility complex MHCI, and screening a polypeptide sequence having the affinity exceeding a specified threshold; andC, predicting polypeptide enzyme digestion sites of the polypeptide sequences encoded corresponding to the mutant genes, and retaining the polypeptide sequences in which the enzyme digestion sites arenot in the polypeptide sequence having the affinity exceeding the specified threshold and which can be separated into segments and cannot be enzyme-digested, as candidate polypeptide sequences of thetumor new antigen. The screening method for the tumor new antigen facilitates subsequent further targeted experiments based on the screening for the tumor new antigen, can greatly reduce the number of the experiments, and saves time, labors and expenditures.

The invention belongs to the field of rapeseed genetic engineering and relates to a gene for controlling a rape plant type, a molecular marker and an application thereof. The gene for controlling a rape plant type can control plant height, branching angles and leaf morphology. Through map-based cloning and genetic transformation, a gene BnaA3. IAA7 for controlling a rape plant type is cloned and verified. The gene is located on an A3 chromosome of a rape genome, codes an auxin protein and has a sequence shown in the formula of SEQ ID NO: 1. The invention also discloses a separated mutant geneBnaA3. Iaa7 of the gene BnaA3. IAA7. The mutant gene has a sequence shown in the formula of SEQ ID NO: 4 and can significantly reduce the plant height and branching angles of rape. The mutant gene does not influence the yield per plant when improving the rape variety or the inbred plant type, and increases the yield per plant when improving the rape hybrid plant type. The invention also disclosesa related molecular marker.