126 results about "Deletion mutation" patented technology

The present application relates to a kind of deletion mutant nucleic acid and its application in anti-herbicide.The deletion mutation occurs in the promoter of rice OsHPPD Gene, the length of deletion sequence is at least 10 bp, and the A in the start codon ATG of gene is 0, at least the bases located in the upstream of start codon ATG from 2041 to 2032 are deleted. OsHPPD The present application finds that by deleting part of the sequence in the promoter of rice OsHPPD Gene, rice can obtain resistance to HPPD inhibitor herbicides, which is of great application value for ensuring agricultural production safety and improving the efficiency of herbicide use.

The invention discloses a genomics cross-modal condition generation method and system based on a potential diffusion model, and belongs to the field of bioinformatics, and the method comprises the following steps: respectively designing and constructing an auto-encoder for gene expression and gene mutation data, and carrying out model training; constructing a potential feature space according to an encoder of gene mutation data and training an unconditionally guided potential diffusion model; constructing a cross-modal feature alignment network according to the two trained encoders and training the cross-modal feature alignment network; and performing joint fine tuning on the unconditionally guided potential diffusion model and the cross-modal feature alignment network. According to the method, the problem of sample modal deletion in multi-modal learning is solved, potential correlation between gene expression data and gene mutation data can be captured through cross-modal feature alignment, information flow between different modals is enhanced, and high-quality prediction data is effectively generated when mutation data is deleted through the potential diffusion model.

The invention relates to a deleted mutant nucleic acid and application thereof in herbicide resistance. The deletion mutation occurs in a promoter of a rice OsHPPD gene, the length of a sequence of a deleted nucleic acid is at least 25 bp, A in an initiation codon ATG of the OsHPPD gene is taken as a 0 site, and at least bases at 2299 to 2275 sites on the upstream of the initiation codon ATG are deleted. According to the application disclosed by the invention, the rice can obtain resistance to HPPD inhibitor herbicides by deleting part of sequences in the promoter of the rice OsHPPD gene, so that the application has great application values in guaranteeing agricultural production safety and improving the use efficiency of the herbicides.

The invention relates to a deleted mutant nucleic acid and application thereof in herbicide resistance. The deletion mutation occurs in a promoter of the rice OsHPPD gene, the length of a deletion sequence is at least 10 bp, A in an initiation codon ATG of the OsHPPD gene is taken as a 0 site, and at least 2041-2032 sites of basic groups located on the upstream of the initiation codon ATG are deleted. According to the application disclosed by the invention, the rice can obtain resistance to HPPD inhibitor herbicides by deleting part of sequences in the promoter of the rice OsHPPD gene, so that the application has great application values in guaranteeing agricultural production safety and improving the use efficiency of the herbicides.

The invention discloses a molecular marker related to thousand seed weight of wheat, a detection primer and application of the molecular marker and the detection primer, and belongs to the technical field of molecular marker-assisted breeding. The nucleotide sequence of the molecular marker TaDTX50-STS is as shown in SEQ ID NO. 1; basic groups from the 502 site to the 661 site of the sequence as shown in SEQ ID NO. 1 have insertion / deletion mutation. The InDel molecular marker related to the thousand seed weight character in a wheat genome is identified and named as TaDTX50-STS, and by detecting the polymorphism or genotype of the InDel molecular marker, the InDel molecular marker can be used for identifying or assisting in identifying the thousand seed weight character of wheat, assisting in breeding dominant varieties with high thousand seed weight of wheat and improving the wheat breeding efficiency.

The invention relates to the technical field of gene editing, and particularly provides a method for creating herbicide-resistant rice through gene editing of an OsEPSPS gene promoter region, and the method is characterized in that WT / 29bp single allelic deletion mutation is introduced into the OsEPSPS gene promoter region, i.e., A in an OsEPSPS gene initiation codon ATG is 0 site, A in the OsEPSPS gene initiation codon ATG is 0 site, A in the OsEPSPS gene initiation codon ATG is 0 site, and A in the OsEPSPS gene initiation codon ATG is 0 site; 29bp is deleted from the 112nd site to the 1150th site at the upstream of the initiation codon ATG on one allele, and the other allele is kept unchanged. By introducing the single allelic deletion mutation, the rice can obtain resistance to EPSPS inhibitor herbicides, the resistance is identified in T0-generation plants carrying the mutation and can be stably inherited to offspring plants, and the application value is high for guaranteeing agricultural production safety and improving the use efficiency of the herbicides.

The invention relates to a T7RNA polymerase mutant for synthesizing single-stranded DNA as well as a screening method and application of the T7RNA polymerase mutant, and belongs to the technical field of enzyme engineering. The invention provides a method for directed evolution and screening of a T7RNA polymerase mutant, which comprises the following steps: knocking out exonuclease and mismatch repair protein from a chassis strain genome, knocking in single-stranded annealing protein, resistance gene containing deletion mutation and a single-stranded DNA synthesis template, and constructing to obtain a genetically engineered bacterium for screening; a T7RNA polymerase mutant library is constructed, recombinant plasmids with the mutant library are transferred into engineering bacteria, T7RNA polymerase mutants synthesize single-stranded DNA to repair deletion mutation of resistance genes, the transformed engineering bacteria can grow on a resistance plate, the T7RNA polymerase mutants obtained through multiple rounds of screening can efficiently synthesize the single-stranded DNA through in-vitro verification, and the T7RNA polymerase mutant can be used for preparing the single-stranded DNA. A brand new tool is provided for synthesis of single-stranded DNA, and the method has the potential of being applied to the fields of gene editing, evolutionary engineering and the like.

The present invention provides a quinazolinone-fused five-membered heterocyclic derivative with a structure represented by formula (I-A). Experimental results show that the compound provided by the present invention has good inhibitory activity against MAT2A, has inhibitory activity against the proliferation of cancer cells with an MTAP deletion mutation, and can be used for treating and / or preventing MAT2A-related cancer diseases.

The invention discloses a rice mutant gene and an identification method. Compared with an OsNRAMP5 gene sequence of an indica rice Shuhui 498 genome, the OsNRAMP5 mutant gene of the rice is subjected to base A deletion mutation at the downstream + 1083 site of an initiation codon ATG. The invention creatively proposes that groups are firstly grouped, grouping screening of OsNRAMP5 gene variants is carried out by utilizing high-throughput targeted sequencing of seed endosperm mixed sample DNA, and then only seeds of a variation group in which the screened variant mixed sample DNA with the highest mutation possibility is located are sown into seedlings. And subsequently screening and identifying single plants carrying corresponding variation in the variation group according to single plants. The technology can greatly save the cost of time, land, reagents and the like, and improves the screening efficiency and land utilization efficiency. The method disclosed by the invention is high in efficiency, good in accuracy, simple and convenient to operate, capable of better realizing high throughput, low cost and operation convenience, and suitable for popularization and application.

The invention discloses a construction method of CiYMaV CRP mutant infectious clone. A CiYMaV infectious clone vector is prepared by inserting a CiYMaV complete sequence into the downstream of a 2 * 35S promoter of a vector pCASS4-RZ; the mutant is obtained by carrying out alanine site-directed mutation or deletion mutation on a CRP (C-reactive protein) region on a CiYMaV infectious cloning vector through overlap extension PCR (polymerase chain reaction), and the site-directed mutation or deletion mutation is obtained on the basis of CiYMaV strong pathogenicity infectious cloning pCiYMaV. According to the invention, a vector pCASS4-RZ is used as a vector skeleton to construct an infectious clone containing strong pathogenicity of CiYMaV, and a series of CRP mutant infectious clone which is coded by viruses and has different pathogenicity and pathogenic expressions are constructed at the same time; virus inoculation experiments show that the CRP protein affects virus accumulation and symptom formation, and a foundation is laid for research on pathogenic mechanisms, virus gene functions and low virulent line cross protection of citrus yellow mottle related viruses and subsequent Mandarivirus virus research.

The invention provides a primer probe set for digital PCR detection of non-small cell lung cancer EGFR mutation and application thereof, and relates to the technical field of digital PCR detection, the primer probe set comprises a primer pair and a probe for specific detection of EGFR gene exon 19 deletion mutation (19del) and exon 21 L858R point mutation, and the 19del mutation probe comprises a plurality of sequences to cover common subtypes. The primer probe group can be combined with a microfluidic digital PCR platform for use, a sample is divided into tens of thousands of microdroplets for amplification, and FAM, HEX, ROX and CY5 fluorescence channel signals are detected to realize absolute quantification. The system provided by the invention has high sensitivity and excellent linearity and precision, is particularly suitable for detecting extremely-low-frequency EGFR mutation in body fluid such as plasma, and has important application value in individualized diagnosis and treatment, curative effect monitoring and drug resistance evaluation of non-small cell lung cancer.

The invention relates to a method and device for simultaneously detecting multiple gene mutations related to thalassemia, and the method comprises the following steps: obtaining nanopore sequencing data of a to-be-detected sample, the to-be-detected sample comprising amplification products of multiple PCR primer pairs for simultaneously detecting multiple gene mutations related to thalassemia; performing quality control screening on the nanopore sequencing data according to the amplification length of the multiple PCR primer pair to obtain first target sequencing data; according to the first target sequencing data, obtaining the sequence number of the amplification product of the PCR multiple primer pair, and according to the sequence number of the amplification product of the PCR multiple primer pair, determining the gene mutation information related to thalassemia of the to-be-detected sample. According to the application, deletion mutation and non-deletion mutation related to thalassemia can be detected at the same time, and the accuracy of HBA1 and HBA2 gene detection results is improved.

The application provides a mussel mucin synthesis method and relates to the technical field of genetic engineering.The application improves the yield of mussel mucin by carrying out deletion mutation on the N-terminal sequence of the secondary structure of the mussel mucin amino acid sequence such as SEQ ID NO.1.The yield of mussel mucin in the prior art is about 2.57g / L, while the yield of mussel mucin in the application is about 5g / L, which is greatly improved compared with the yield of mussel mucin in the prior art.On this basis, the mussel mucin is modified by dopa through co-expression of tyrosinase, and the expression amount of the tyrosinase is controlled through different intensities of RBS, so as to regulate the modification degree of dopa, improve the yield of mussel mucin, and realize controllable modification of dopa in the mussel mucin, finally obtaining mussel mucin with high yield and different modification degrees of dopa.

The invention belongs to the technical field of biological detection, and provides a kit for detecting EGFR (epidermal growth factor receptor) 19del subtypes. According to the invention, by designing four universal upstream primer target segments and four probes combined with the upstream primers, different subtypes of EGFR19del can be accurately detected by a Tm value; by changing the fluorophore combination of the probe, single-tube multi-channel accurate typing can be realized. The primer tag segment and detection probe combination developed by the invention is not only suitable for deletion mutation, but also suitable for detection of fusion mutation and pathogenic microorganisms, and has wide applicability.

A primer pair, method, and use for rapidly identifying newborn nude mice. On the basis that a deletion mutation occurs in the base G at position 337 of a coding region of the Foxn1 gene in nude mice, a primer pair introducing an enzyme digestion site is designed according to sequence characteristics. Using the genome of a mouse to be tested as a template, the Foxn1 gene is amplified according to a PCR-RFLP method. The PCR product is then digested with a SmaI restriction endonuclease, and the digested product is genotyped by agarose gel electrophoresis. In addition, a reaction system that can be combined with lateral flow technology is screened, thereby achieving instant detection of the newborn nude mice. The method has the characteristics of simple operation, short time consumption, low cost, instant detection, etc.

This invention discloses transcription factors As NF-YB3 Its application in regulating drought resistance in oats falls under the field of biotechnology. (Transcription factors) As NF-YB3 The amino acid sequence is shown in SEQ ID NO.1. As NF-YB3 The CDS sequence of the gene is shown in SEQ ID NO.2, and the gDNA sequence is shown in SEQ ID NO.3. Overexpression As NF-YB3 The gene significantly enhances the drought resistance of oats, while also increasing the activity of CAT and SOD enzymes in oats, slowing down the water loss rate of detached leaves, and improving seedling survival rate under drought stress. AsNF-YB3 forms a heterotrimeric complex with AsNF-YA1 and AsNF-YC5, and its 4bp insertion / deletion mutation in the promoter region regulates oat drought resistance by affecting AsARF1 binding and gene expression. This provides a new gene resource and technical pathway for agricultural stress-resistance breeding.

The application discloses a method for improving the protein secretion capacity of Aspergillus and a recombinant strain obtained from the method. An16g07795 The method comprises performing genetic operation on Aspergillus to delete or reduce the activity of specific genes An16g07795 , so as to improve the protein secretion capacity of saccharifying enzyme, alpha-amylase, other proteins participating in starch hydrolysis and the like. The application utilizes the CRISPR-Cas9 genome editing technology to knock out the genes of Aspergillus niger, obtains a gene editing deletion mutant strain, and significantly improves the production level of starch hydrolysis enzymes, including the protein secretion level and the activity of saccharifying enzyme, so the application has great application value.

The invention belongs to the technical field of animal breeding, and particularly relates to application of a detection reagent of a molecular marker related to the hair length character of a down producing goat. The molecular marker is located at 155, 344, 701-155, 346 and 600 positions on a No.1 chromosome of a goat genome, and the nucleotide sequence of the molecular marker is as shown in SEQ ID NO. 1. It is found that the wool length of an individual with a normal molecular marker area is larger than that of an individual with a deletion molecular marker area, and therefore the cashmere goat variety with the large wool length can be bred by detecting deletion mutation of the molecular marker on the chromosome 1 of the goat genome.

This invention relates to the field of gene editing technology, specifically providing a method for creating herbicide-resistant rice by gene editing the promoter region of the OsEPSPS gene. The method involves introducing a WT / -29bp monoallelic deletion mutation into the OsEPSPS gene promoter region. Specifically, with position A of the OsEPSPS gene start codon ATG set to 0, a 29bp deletion is made in one allele located at positions 1122 to 1150 upstream of the start codon ATG, while the other allele remains unchanged. This invention demonstrates that by introducing this monoallelic deletion mutation, rice can acquire resistance to EPSPS inhibitor herbicides. This resistance was identified in the T0 generation plants carrying this mutation and can be stably inherited by their offspring, possessing significant application value for ensuring agricultural production safety and improving herbicide application efficiency.

The present invention discloses a method for constructing a genetically engineered bacterium capable of efficiently synthesizing lipopolysaccharide and its application, belonging to the technical field of genetic engineering. The genetically engineered bacterium of the present invention, E. coli W3110△npr△yhbJ△fabF△P BAD(lpxD ‑fabZ‑lpxA‑lpxB), has undergone deletion mutations of three genes, npr, yhbJ, and fabF, and at the same time, a promoter P has been inserted in front of the chromosomal gene cluster lpxD‑fabZ‑lpxA‑lpxB sequence BAD . The genetically engineered bacterium of the present invention has no resistance markers, has good growth conditions, can be adapted to large-scale production, and the yield of synthesized lipopolysaccharide is 13.0 mg / g DCW, which is 91.2% higher than that of the wild-type E. coli W3110.

The invention belongs to the technical field of gene editing and disease model construction, and particularly relates to a construction method and application of a Wilson disease animal model. Aiming at a large fragment deletion mutation type which exists clinically in Wilson disease but lacks a corresponding animal model, a CRISPR / Cas9 gene editing technology is utilized, a pair of sgRNAs is specifically designed, an eighth exon region of a mouse Atp7b gene is precisely cut and deleted, and a WD mouse model with deletion mutation (c.2333340delGACGGTGG) of eight basic groups of the eighth exon of the Atp7b gene is successfully constructed. The method can be used for research on pathogenesis of WD, screening of novel diagnostic markers and evaluation of curative effect of therapeutic drugs, is particularly suitable for preclinical evaluation of gene therapy strategies and development of adjuvant therapeutic drugs of targeted NLRP3 inflammasomes, and has great scientific research value and clinical transformation prospect.

A method for collecting data for cancer diagnosis, onset risk determination, malignancy determination, and / or prognosis prediction according to the present invention comprises (a) a step for causing an antigen-antibody reaction by bringing a sample from a subject into contact with a tumor-associated antigen; (b) a step for collecting data for cancer diagnosis, onset risk determination, malignancy determination, and / or prognosis prediction, (c) a step for comparing the measured amount of anti-p53 antibody with a predetermined reference amount of anti-p53 antibody for a tumor-associated antigen, and (c) a step for measuring the amount of anti-p53 antibody in a sample that specifically binds to the tumor-associated antigen; the tumor-associated antigen includes one or more p53 isomer mutants comprising an amino acid sequence having a mutation in any one of amino acid sequences of SEQ ID NO: 1-3, and the mutation includes any one of an N-terminal deletion mutation, a C-terminal deletion mutation, and a deletion mutation of both the N-terminal and the C-terminal.

The invention discloses a primer group, a kit and a detection system for thalassemia gene detection. In order to more comprehensively and accurately detect point mutation, small insertion, small deletion, gene fusion mutation, large fragment deletion and gene structure variation on a thalassemia core gene and a modified gene in single detection, the invention provides a thalassemia gene detection primer group and a kit. Based on third-generation sequencing platforms such as a single-base nanopore sequencing platform and the like, all point mutation, small insertion, small deletion and gene fusion mutation in the amplification range of the thalassemia core gene and the modified gene can be detected by utilizing the primer group or the kit; and 60 alpha large fragment deletion mutations, 60 beta large fragment deletion mutations and 7 gene structure variations. The method is beneficial to rapid and comprehensive screening of thalassemia, and is high in detection sensitivity and good in accuracy.

The invention relates to the technical field of gene editing, and particularly provides a method for creating fluazifop-p-butyl herbicide resistant rice through gene editing of an OsACCase gene promoter region. The method comprises the following steps: constructing a crRNA plasmid library covering an OsACCase gene promoter region on the basis of a CRISPR / LbCas12a system, performing agrobacterium-mediated transformation on rice callus, and performing resistance screening, fluazifop-p-butyl herbicide screening and molecular identification to obtain a fluazifop-p-butyl resistant rice strain. The strain is subjected to-4bp / -5bp double allelic deletion mutation in an upstream 1865-site region of an initiation codon ATG of the OsACCase gene, and a homozygous mutant strain and a double allelic deletion mutant strain can normally grow after being sprayed with 6 times of field recommended dose of fluazifop-p-butyl. The herbicide resistance is realized by editing the gene promoter region, the growth defect of coding region mutation is avoided, the breeding technology is enriched, and weed prevention and control are assisted.

The invention relates to a T7RNA polymerase mutant for synthesizing single-stranded DNA as well as a screening method and application of the T7RNA polymerase mutant, and belongs to the technical field of enzyme engineering. The invention provides a method for directed evolution and screening of a T7RNA polymerase mutant, which comprises the following steps: knocking out exonuclease and mismatch repair protein from a chassis strain genome, knocking in single-stranded annealing protein, resistance gene containing deletion mutation and a single-stranded DNA synthesis template, and constructing to obtain a genetically engineered bacterium for screening; a T7RNA polymerase mutant library is constructed, recombinant plasmids with the mutant library are transferred into engineering bacteria, T7RNA polymerase mutants synthesize single-stranded DNA to repair deletion mutation of resistance genes, the transformed engineering bacteria can grow on a resistance plate, the T7RNA polymerase mutants obtained through multiple rounds of screening can efficiently synthesize the single-stranded DNA through in-vitro verification, and the T7RNA polymerase mutant can be used for preparing the single-stranded DNA. A brand new tool is provided for synthesis of single-stranded DNA, and the method has the potential of being applied to the fields of gene editing, evolutionary engineering and the like.

The present invention relates to improved microbial enzymes with transfructosylation activity for efficient and cost-effective production of fructo-oligosaccharides. More specifically, the invention is directed towards obtaining mutant FTase family of genes from genus Aspergillus. Nucleic acids, peptide sequences, mutant proteins, vectors and host cells for recombinant expression of novel FTases are also provided. Various mutations, such as but not limited to point mutations and deletion mutations as well as combinations thereof are presented herein. The invention also relates to a process for the expression of a novel recombinant FTase mutants as a secreted protein. The enzymes exhibit high purity after filtration, which eliminates the need for costly chromatographic procedures.

The invention relates to the technical field of gene editing, and particularly provides rice OsEPSPS gene promoter region deletion mutation and application thereof in creation of herbicide-resistant rice, the deletion mutation is that a sequence with the length of 22bp or 28bp is deleted in the OsEPSPS gene promoter region, namely, A in an OsEPSPS gene initiation codon ATG is 0 site, A in the OsEPSPS gene initiation codon ATG is 0 site, A in the OsEPSPS gene initiation codon ATG is 0 site, and A in the OsEPSPS gene initiation codon ATG is 0 site; and deleting the basic groups at sites 2534 to 2555 upstream of the initiation codon ATG or the basic groups at sites 2527 to 2554 upstream of the initiation codon ATG. According to the application disclosed by the invention, the rice can obtain resistance to EPSPS inhibitor herbicides by deleting part of sequences in the promoter of the rice OsEPSPS gene, so that the application has great application values in guaranteeing agricultural production safety and improving the use efficiency of the herbicides.

The invention provides a primer group, a method and a kit for detecting thalassemia, which can be used for simultaneously detecting thalassemia related genes, related regulatory regions and modified gene mutations. According to the method, multiple regions and multiple mutations of HBA1, HBA2, HBB and other genes, related regulatory regions (LCR, 3 'HS, HS-40 and MCS-R) and modified genes (HBD, MYB, BCL11A and KLF1) are detected at the same time based on multiple PCR amplification and three-generation sequencing, amplification of related gene deletion type and non-deletion type mutations can be achieved at the same time in a single reaction tube through multiple PCR; a third-generation sequencing platform (an ONT sequencing platform and a PacBio company sequencing platform) is subsequently adopted for subsequent detection, the method is easy and convenient to operate, multiple PCR and a third-generation sequencing library are reliable in quality and high in repeatability, application of the third-generation sequencing technology to clinical detection is facilitated, and the method is suitable for clinical detection. According to the invention, various deletion and non-deletion mutation, complex structure variation and the like can be detected at the same time; when two or more mutations exist on the HBA1 / 2 or HBB gene locus at the same time, cis-mutation or trans-mutation can be distinguished.

The present invention contemplates-allele-specific gene editing based on targeting a heterozygous single nucleotide polymorphism (SNP) in a protein coding sequence associated with a genetic disease. The data shown herein demonstrates that the outcome of such gene editing creates a nonesense mutation that results in a marked and selective reduction of mutant protein without affecting wild type protein expression. Expression of a single CRISPR-Cas9 nuclease in neurons generated a high frequency of mutations in the targeted HD allele that included both small insertion / deletion mutations and viral vector insertions. Thus, as disclosed herein, allele-specific targeting of InDel and insertion mutations to heterozygous coding SNPs provides a feasible approach to inactivate autosomal dominant mutations that cause genetic disease.

The invention relates to the technical field of biology, in particular to a primer probe set for detecting mutation of a bZIP region of a CEBPA gene, a kit and a drop-offdPCR method. The primer and the probe are designed according to the DNA sequence of the CEBPA gene, and cover the area where common in-frame mutation sites in the bZIP area of the gene are located. When mutation exists in the bZIP region of the CEBPA gene, the wild-type probe located at the mutation site cannot be tightly combined with the template, and the other three wild-type probes located outside the mutation site in the same group can still be combined with the template, so that mutation of multiple sites can be detected through one experiment. The kit disclosed by the invention can be used for detecting in-frame insertion / deletion mutations including K304Q305insL, Q305dup, K313V314insE and the like in the bZIP region of the CEBPA gene, has relatively high sensitivity, and can be used for MRD (Magnetic Resonance Diffraction) monitoring.