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142 results about "Mutation screening" patented technology
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Mutation Screening. Mutation Screening is a tool for couples to identify if they are carriers of a heredity disease. Mutation Screening by Sanger Sequencing is most often recommended when the genetic disease has previously been identified or there is a known family history for a particular genetic disease.
A (Rhizopus Oryzae)ME-F11, its mutationscreening method and method for fermenting fumaric acid are disclosed. The preserving code is CCTCC NO: M 207066. The fumaric acid concentration reaches to 90-120g / L, total yield reaches to 90-95 wt% . It's simple and can be used for industrial production.
The invention discloses a combined primer for screening human deafness genemutation. The combined primer comprises a combined forward primer and a combined reverse primer, wherein the combined forward primer consists of two parts namely a joint part P1 sequence and a forward primer sequence, and the combined reverse primer sequentially consists of a joint part P2 sequence, a label sequence and a reverse primer sequence. The invention also discloses a kit for screening the human deafness genemutation. The invention also discloses applications of the combined primer and the kit in preparation of a reagent for screening the human deafness multi-genemutation. A primer containing the label (Barcode) sequence is used for differentiating different samples, and at least 32 samples can be detected by performing a method provided by the invention once, so that combined primer can be widely applied to general investigation of deafness susceptible genes. The price of the combined primer disclosed by the invention is far lower than that of imported instruments or reagents, so that the sequencing cost is greatly reduced and then the detection cost is also greatly reduced.
The invention discloses a human K-ras (K-rat sarcoma) genemutation parting type fluorescent quantitation PCR (polymerasechain reaction) detecting reagent kit and a detecting method. The detecting reagent kit comprises PCR mixed reaction liquid, a peptidenucleic acid probe, a taqman-MGB (minor groove binder) probe, an inner control forward and reverse primer and probe, an external control forward and reverse primer and probe and a positive reference article, wherein the inner control forward and reverse primer and probe is used for K-ras genemutation detection, and the external control forward and reverse primer and probe is used for the K-ras genemutation detection. The PNA (peptidenucleic acid) technology and the taqman-MGB technology are combined, relevant ARMS (amplification refractory mutation system) primers are designed, PNA can be stably combined with wild K-ras genes 12 or 13 codon sequences, only mutation specimens can be amplified, and the K-ras gene mutation can be detected. The method has the advantages that the speed is high, simplicity and convenience are realized, the specificity is good, the sensitivity is high, and the method can be used for the clinical K-ras gene mutation screening.
The invention relates to primers and probes for detecting human K-ras genemutation as well as use method thereof, relating to detection of genemutation. In the invention, the fluorescent quantitative PCR technology and the fluorescence dual ring probes technology are combined to develop a multiple real-time fluorescent PCR method capable of fast detecting genemutation and design the relevant primers. The method is used for detecting mutation of the twelfth and the thirteenth codons of the K-ras gene, mutant type primers designed specific to different mutation types are respectively adopted to detect samples to be detected, only the mutation type samples can be smoothly amplified to obtain double-strand DNA products which can be combined with DNA probes to send out fluorescence signals to be detected. The invention can detect mutate type genes with content being 1 / 1000 in the samples to be detected, has the advantages of high speed, convenience and simplicity, safety, high sensitivity, high throughput and low cost and can be applied to K-ras gene mutation screening of numerous clinical samples.
The invention discloses organophosphorus degrading enzyme mutants and coding genes and application thereof. Site-directed mutation screening is performed on original enzyme coding genes to obtain three organophosphorus degrading enzyme mutants according to the spatial structure characteristic of organophosphorus degrading enzyme OPHC2 by computer-aid molecular design, and the amino acid sequences of the three mutants are shown by SEQ ID NO.2, SEQ ID NO.4 and SEQ ID NO.6 respectively. Compared with the original enzyme, the organophosphorus degrading enzyme mutants improve the ethyl-parathion degradation specific activities by 2.45 times, 2.56 times and 4.51 times, and the degradation capabilities (kcat / Km value) of the organophosphorus degrading enzyme mutants are 2.7 times, 3.3 times and 4.2 times that of the original enzyme. Enzymatic property analysis shows that the enzymatic property of the organophosphorus degrading enzyme mutants is not changed compared with the original enzyme, and the both have good heat resistance.
The invention relates to bacillus mucilaginosus and application thereof for preparing a salinealkali soil improvement agent. The bacillus mucilaginosus JDzhs-bh is preserved at the China Center for Type Culture Collection on March 20th, 2019, the address is No.3, courtyard 1, the western Beichen road, Chaoyang district of Beijing, and the strain preservation number is CGMCC No.17376. The invention discloses bacillus mucilaginosus capable of resisting high temperature and saline alkali and being cultivated through ultravioletmutation screening for the first time. The bacillus mucilaginosus has the stress resistance for high temperature and saline alkali, and can effectively adapt to saline alkali hardening soil. Meanwhile, the effect of degrading silicon and phosphorus is achieved, release of elements such as soluble silicon and phosphorus in soil is promoted, cropstress resistance is enhanced, the utilization rate of the soil repairing agent is achieved, saline alkali soil is repaired and improved, and salinization is lowered.
The invention provides an exome potential pathogenic mutation detection method based on a family line. The detection method comprises the following steps: 1) reading a result file of an exomesequencing dataprocessing flow, and conducting function filtering; 2) reading the file obtained in the last step, extracting mutations in all samples, calculating a union set, and then combining all samples, so that a matrix is constituted; 3) extracting mutation information in the matrix obtained in the last step, enumerating and assessing pathogenicity of single mutation and pathogenicity of combined dual-site mutation, so that a potential pathogenic mutationlist is obtained; and 4) in accordance with the list obtained in the last step, calculating the appearance situations of sites in various samples and target genes. According to the method disclosed by the invention, data integration and basic filtration are completed by taking an output result of the common exome sequencing processing flow as an input condition; by virtue of a special mutationscreening algorithm, a candidate set of the potential pathogenic mutations is provided; and the method focuses on solving a problem on potential pathogenic mutation mining of sequencing data with high heterogeneity, high mutation rate and high noise.
The invention relates to a strain for producing L-lysine, which is classified and named as Escherichiacoli NT1003 delta Met delta Thr and is collected in China Center for Type Culture Collection (CCTCC) on May 30, 2013, and the collection number is CCTCC NO: M2013239. The invention further provides a mutationscreening method of the strain and a method for producing the L-lysine by utilizing the strain. After continuous passage is performed on the Escherichiacoli NT1003 delta Met delta Thr strain for 7 times, the content of the L-lysine in a fermentation solution is about 60g / L and has no significant change, so that the genetic stability is good.
The invention discloses a kit for synchronously detecting 12 mutation hotspots of the Chinese population phenylketonuria PAH (phenylalanine hydroxylase) gene. In the kit, 12 loca on the Chinese population PAH gene are used as detection objects; an amplification primer and an extension primer are respectively designed for the mutation type of each locus; each destination section is subjected to PCR amplification and mark extension at the same time; and the gene types of the 12 mutation hotspots are obtained at the same time through capillary electrophoresis analysis. The invention provides a phenylketonuria PAH gene mutation screening kit which is simple, has the advantages of high flux, high performance, low cost and high detection accuracy, is suitable for the Chinese people and suitable for the group screening of the Chinese population phenylketonuria.
The invention discloses a universal strategy for efficiently improving enzymethermodynamics stability. The strategy is specifically implemented in the way of taking a catalytic residue of zymophore as the center, selecting a high-flexibility residue 10 angstroms away from the center at most as a target locus according to the B-factor value in an enzymecrystal structure, and obtaining a mutant with enzymethermodynamics stability improved through mutation screening. Under the guidance of the strategy, the thermal stability of candida rugosalipase 1 with a highly complicated structure is improved remarkably. By means of stability change results of protein (LipA) with the smallest structure and moderately complicated protein (CalB), it is found that the success rate of enzyme mutation can be increased remarkably through mutation of residues with high B-factor 6-10 angstroms away from the catalytic residue. Meanwhile, through system analysis of a mutant structure-function relationship, a foundation is laid for deep research of enzyme region stabilization law and development of enzyme stabilization technique.
The invention relates to a strain for generating phospholipase D with high and stable yield by utilizing physical and chemical mutation, belonging to the technical field of microbial mutation. The strain is characterized in that the phospholipase D is prepared by fermenting streptomyces chromofuscus of high yield phospholipase D, which is obtained by taking the streptomyces chromofuscus as an original strain and sequentially carrying out composite mutation of ultraviolet rays and lithiumchloride and composite mutation of atmosphere cold plasmas and the lithiumchloride on a monospore bacterial suspension thereof. The invention has the advantages that the streptomyces chromofuscus of the high yield phospholipase D is obtained through mutation screening, thereby providing a high yield strain with higher application value to industrial production of the phospholipase D, laying a good foundation for fermentation culture and application of the phospholipase D, and also providing a good and feasible method for microbial mutation breeding. The invention adopts a mutation and screening method, has the advantages of simple operation, high efficiency and the like, and has a practical value on screening the strain with an industrial application value.
The invention relates to POU3F4 mutantgene which has 499C> T mutation, which is relative to human genetic deafness. The invention provides a reagent kit for detecting POU3F4 gene 499 C>T mutation, and a detection process which is used to diagnose the reason and kind of deafness generation through detecting whether patients have the mutation genes. The mutantgene and the detection process are beneficial for the POU3F4 mutation screening work of deafness patients which is carried out on clinics, and provides services for the diagnosis and treatment of deaf patients.
The invention discloses an aureobasidium pullulans for highly producing pigment-free Pulullan, which is collected in China Center For Type Culture Collection (CCTCC) with a collection number of CCTCC M2013611. The invention further discloses a mutationscreening method and a culture method of aureobasidium pullulans for highly producing pigment-free Pulullan, and a method for producing Pulullan through fermentation of the aureobasidium pullulans for highly producing pigment-free Pulullan. A culture obtained by fermentation cultivation of an aureobasidium pullulans strain GM-1 cannot show a dark green color or deep green color due to accumulation of melanin substance, but form a milk white or faint yellow composition. Compared with a conventional Pullulan producing strain, the aureobasidium pullulans strain GM-1 has the advantages that the Pulullan is colorless and is higher in yield, the purposes of increasing the raw materialutilization rate and reducing the production cost are reached, and the demand of application fields such as foods, medicines, and environment on the pigment-free Pulullan is met.
The invention discloses a white rot Inonotus hispidus mutant strain T906 for laccase production and a preparation method thereof. The mutant strain T906 (CoriolopsisgallicaT906) mentioned in the invention is preserved in China Center for Type Culture Collection. The preservation number is CCTCC NO: M2011317, the preservation date is September 15, 2011 and it is preserved in Wuhan University, Wuhan, China. Wild Inonotus hispidus (preservation number being cfcc88387) purchased from China Forestry Culture Collection Center is used as an original strain, and the mutant strain is obtained by ultraviolet and nitrosoguanidine combined mutation screening. Under an optimum condition, the laccase production of the mutant strain reaches 213U / mL. The mutant strain has mycelial morphology and cultural characteristics which are obviously different from the original strain, does not degenerate after serial passages for over 30 times and has genetic stability.
The present invention discloses a congenital adrenal hyperplasiagene screening kit. The congenital adrenal hyperplasiagene screening kit comprises a PCR amplification primer mixed liquor used for detecting 17 mutation sites including CYP 21A2 gene; an extended primer mixed liquor for the above 17 mutation sites; dNTP; 10*RCR buffer; 25 mM Mg2+ ions; Fast StartTaq enzyme; purification reagent composed of SAP enzyme, Exon I enzyme and a matched buffer; SNaPshot Multiplex mixed liquor; single-site homozygous and heterozygous mutation positive and negative controlDNA; and deionized water. According to the technical scheme of the invention, a large number of researches and practices show that, the mutation sites of the congenital adrenal hyperplasia gene in the Chinese population are screened and combined. By utilizing the kit, 17 mutation sites of the specific congenital adrenal hyperplasia gene are simultaneously subjected to multiplex nested PCR amplification, labeling and extending, so that the defects of the existing mutation screening method are overcome. The kit is simple in operation and low in cost. The detection throughput and the accuracy of the detection result are greatly improved compared with the prior art.
The invention belongs to the technical field of biological engineering and synthetic biology, and particularly relates to a multi-fragment deoxyribosenucleic acid (DNA) series connection recombination assembly method based on BT1 integrase and mutation recognition sites thereof. In the method, sixteen pairs of mutational integrase recognition sequences are provided through mutation screening and verification, on the basis, a complete set of related carriers is constructed and comprises a framework carrier and seven TA cloning vectors, the framework carrier can realize escherichia coli-streptomyces coelicolor shuttle, the seven TA cloning vectors comprise mutation substrates and are used for DNA element construction and screening, and a concrete method for multi-fragment external series connection recombination reaction is provided. The method of the invention is simple, fast and efficient and can be used for the multi-fragment fast series connection recombination assembly, and the amplification of series connection products is realized through escherichia coli transformation. The method can be widely used for combination and assembly of standard modules and elements in the synthetic biology.
This disclosure teaches high throughputmutation screening methods allowing simultaneous analysis of multiple genetic regions of interest and sensitive detection of very low frequencymutation(s) by the use of a universalized approach. Methods comprise treating RNA:DNA heteroduplexes of interest with a ribonuclease, sequence extension by an RNA-primed DNApolymerase, ligation with a blocking adapter, and differential sequence fill-in followed by single-strand-specific nucleasedigestion to permit full-length sequence extension and subsequent ligation with a tagged reporter adapter solely in mutants filled in with a complementary deoxyribonucleotide triphosphate. By forming tagged mutant-dual adapter hybrids or mutant-triple adapter hybrids, the detection and / or quantification of mutants may be directed to the commonly shared tag(s) or flanking adapter sequences for signal detection / enhancement or sequence amplification in all different mutants regardless of the source or the number of mutations involved, thereby avoiding the tremendous effort of multiple target-specific sequence amplifications. Methods may be performed wholly or partially in solution, on solid phase media, in large scale, adapted for automated or semi-automated analysis, and any combinations thereof.
The invention discloses a detection method for mutation screening of disease-causing genes of Parkinson's disease (PD). The detection method comprises the following steps: (1) adding dd H2O into a synthesized MIPs probe to prepare a probe solution with the concentration of 100mu M; (2) sucking 1mu l of the probe solution in step (1) by utilizing a pipettor; then putting the probe solution into a 1.5ml new EP pipe; sufficiently and uniformly mixing; marking a pipe wall as mixed-MIPs by utilizing a Marker pen; (3) preparing an MIPs probe phosphorylation activation system and carrying out activation on the mixed-MIPs; (4) diluting a mixed-MIPs probe; (5) preparing an MIPs target capturing reaction system; (6) preparing an MIPs enzymedigestionreaction system; (7) carrying out PCR (PolymeraseChain Reaction) amplification reaction; (8) carrying out polyacrylamidegel electrophoresis and magnetic bead purification; (9) carrying out high-throughput sequencing on a mixed library; (10) carrying out Sanger sequencingverification. By adopting the detection method for the mutation screening of the disease-causing genes of the Parkinson's disease, the mutation screening of the 18 PD disease-causing genes of the Parkinson's disease can be finished, and the detection method has important significance on researches of super-early diagnosis and PD pathogenesis of potential PD patients.