18 results about "Gene deletion mutation" patented technology

The invention relates to a Pseudomonas plecoglossicida pldB gene deletion mutant strain and a preparation method and application thereof, and belongs to the technical field of microorganisms and the field of algae preservation, the pldB gene of the mutant strain is deleted or inactivated, the nucleotide sequence of the pldB gene is shown as SEQ ID NO: 1, and the GenBank accession number of the Pseudomonas plecoglossicida is CP031146.1. The virulence of the mutant strain is obviously reduced compared with that of a wild strain of pseudomonas plecoglossicida; the strain can induce appropriate immune response of a host and up-regulate expression of anti-inflammatory factors, is beneficial to relieving immune pathological injury, has good attenuated live vaccine potential, and provides a brand new vaccine candidate strain for prevention and control of the visceral white-spot disease of the large yellow croaker.

The application discloses a rice OsNAR1 application of the gene or the coded protein in regulating rice resistance to Magnaporthe oryzae, and relates to the technical field of rice Magnaporthe oryzae prevention and treatment. OsNAR1 The CDS nucleotide sequence of the gene is shown as SEQ ID No. 2. OsNAR1 The application finds that the rice gene OsNAR1 plays an important role in the process of rice resistance to Magnaporthe oryzae, the number of leaf spots on the gene deletion mutant is less, the gene can be used for screening rice strains resistant to Magnaporthe oryzae, the expression amount of a basic resistance marker gene in the gene deletion mutant is significantly up-regulated, and it is indicated that OsNAR1 negatively regulates the expression of the basic resistance gene and can be used for preventing and treating Magnaporthe oryzae.

The application belongs to the field of genetic engineering and medical engineering, and discloses virK Application of gene in regulation of Klebsiella pneumoniae virulence. The application finds that the expression level of the gene is significantly increased in polymyxin-resistant mutant strains through transcriptome analysis, virK Further research shows that the expression of the gene is regulated by the PhoP / PhoQ two-component regulatory system. The application constructs virK Gene deletion mutant strains and complementary strains, and compares the virulence difference of the strains in an animal infection model, finds that virK The virulence of the deletion strain is significantly reduced, and the complementary strain can restore the virulence phenotype, indicating that virK The gene plays an important role in the regulation of Klebsiella pneumoniae virulence. The application first finds that PhoP can regulate virK Gene expression, and affects the virulence phenotype of Klebsiella pneumoniae through the regulation pathway, thereby revealing a new virulence regulation mechanism, and providing a theoretical basis for subsequent targeted intervention.

The present application relates to the field of biotechnology, in particular to the application of glycoside hydrolase FsGH28c and its coding gene in regulating pathogenicity of a bacterial strain. The present application provides a glycoside hydrolase coding gene FsGH28c related to the development of Fusarium solani pisi, the utilization of carbon source and pathogenicity. After knocking out the FsGH28c gene of Fusarium solani pisi, the spore production of Fusarium solani pisi is reduced, the spore development is deformed, and the utilization rate of carbon source (especially pectin) is reduced. In addition, the amount of fungal colonization at the root of the host pepper and the pathogenicity of the FsGH28c gene deletion mutant are reduced. The FsGH28c gene of the present application positively regulates the pathogenicity of Fusarium solani pisi, and can be used as an important candidate gene for the study of the pathogenic mechanism of Fusarium solani pisi.

This invention discloses a *Lactobacillus paracasei* S-NB gene deletion mutant strain, its construction method, and its application. The S-NB gene deletion mutant strain uses *Lactobacillus paracasei* S-NB as the starting strain, and the mutant strain contains *Lactobacillus paracasei* S-NB gene deletion mutants. cps The upstream segment of the gene was knocked out by the plasmid. cps Upstream homologous arm gene substitution in *Lactobacillus paracasei* S-NB cps The downstream gene fragment carried by the knockout plasmid cps Gene substitution in the downstream homologous arm of the gene; among which, cps The gene base sequence of the upstream homologous arm is shown in SEQ ID NO:1. cps The downstream homologous arm gene base sequence is shown in SEQ ID NO:2. Compared with traditional methods, the success rate of knockout vector construction in this invention reaches over 95%, and the operation is simple, time-saving, and the operation cycle is only 6-7 days; the knockout vector constructed using this invention can knock out Lactobacillus paracasei S-NB. cps Genes, the result cps Gene deletion mutants are unaffected in growth, but their capsule layer becomes thinner and their ability to produce biofilm decreases.

The invention provides an application of a CsSTE12 gene in inhibition of Philospora piricola. The nucleotide sequence of the CsSTE12 gene is shown as SEQ ID NO.1, and the coding amino acid sequence of the CsSTE12 gene is shown as SEQ ID NO.2. The invention also provides an application of the CsSTE12 gene in inhibition of Philospora piricola in inhibition of Philospora piricola. According to the CsSTE12 gene deletion mutant disclosed by the invention, the CsSTE12 gene is knocked out, so that the obtained CsSTE12 gene deletion mutant cannot form a normal spore structure, and spores cannot fall off from hyphae, and the CsSTE12 deletion is proved to regulate the morphology of the spores of the helminthosporium basicola and influence the production of the spores. Molecular exploration is carried out from the perspective of pathogenic bacteria, and the application has important guiding significance in research on prevention and treatment of plant diseases caused by wheat root rot helminthosporium.

The present application belongs to the field of plant disease control research, and relates to a cucumber multi-host coelomyces effector protein CcSP1, a coding gene thereof and application. The present application provides a new cucumber multi-host coelomyces effector protein CcSP1, a coding gene thereof and application. The nucleotide sequence of the coding gene of the multi-host coelomyces effector protein CcSP1 is shown as SEQ ID NO. 1. Transient expression of the CcSP1 gene in N. benthamiana can inhibit hypersensitive necrosis caused by N. benthamiana. A gene deletion mutant ΔCcSP1 is constructed by homologous recombination, and the deletion of the gene does not affect the vegetative growth, but significantly reduces the pathogenicity when invading the cucumber leaves. It is shown that CcSP1 is involved in the lesion expansion of the plant after the invasion of the multi-host coelomyces, and plays an important role in the pathogenic process of the multi-host coelomyces. The present application has important significance for analyzing the related molecular mechanism of cucumber coelomyces leaf spot and establishing the comprehensive control technology strategy of the plant cucumber coelomyces leaf spot.

The invention relates to the technical field of molecular biology, in particular to a preparation method of a pabpc1a gene deleted zebrafish mutant, which is used for preparing the pabpc1a gene deleted zebrafish mutant. According to the preparation method, the CRISPR / Cas9 technology is utilized for the first time to perform targeted knockout on the pabpc1a gene in the zebra fish, and the pabpc1a gene deletion mutant capable of being stably inherited is obtained. The zebrafish mutant with the deletion of the pabpc1a gene, which is constructed and obtained by the invention and can be stably inherited, is expected to be applied to research on the function of the pabpc1a gene, and is expected to be applied to research on an embryonic development molecular mechanism and high-throughput drug screening.

The invention belongs to the technical field of biological medicines, and discloses an induced pluripotent stem cell (iPSC) capable of retaining NFIA gene deletion mutation and application of the iPSC. The stem cell strain is preserved in the China Center for Type Culture Collection (the preservation number is CCTCC NO: C202590), carries NC000001.10: g.6165096761842967del heterozygous deletion mutation, contains deletion of exons 3-6 of an NFIA gene, stably expresses pluripotent markers (OCT4, SOX2, NANOG and SSEA4), and has the capability of differentiating to three germ layers. The preparation method comprises the following steps: separating PBMC (peripheral blood mononuclear cells) from peripheral blood of a patient, carrying out reprogramming by utilizing 2.0 Sendai Reprogam Kit, and amplifying pluripotent clone in Essental 8TMMedium. The stem cell strain can be used for constructing a research model of neurodevelopmental disorder diseases (such as corpus callosum insufficiency, hydrocephalus and development retardation), simulating pathological phenotypes by differentiating neurons, glial cells or brain organs, and is also suitable for drug screening and gene therapy research. And a precise humanized tool is provided for mechanism analysis and treatment development of NFIA deficiency related diseases.

The invention provides a primer group and a kit for simultaneously detecting IKBKG gene and IKBKGP1 gene Exon4-10 deletion mutation and application of the primer group and the kit, and belongs to the technical field of gene detection. The primer group disclosed by the invention comprises a primer 1 with a nucleotide sequence as shown in SEQ ID NO.1, a primer 2 with a nucleotide sequence as shown in SEQ ID NO.2, a primer 3 with a nucleotide sequence as shown in SEQ ID NO.3 and a primer 4 with a nucleotide sequence as shown in SEQ ID NO.4. According to the invention, through one-time amplification detection, whether the gene sequence of IKBKG has the Exon4-10 deletion or the gene sequence of IKBKGP1 has the Exon4-10 deletion can be clearly known. For a sample with positive Exon4-10 deletion, the type of the IKBKG and / or IKBKGP1 subjected to the Exon4-10 deletion mutation can be clearly known.

The present invention discloses a phage genome editing vector based on the CRISPR-Cas9 system and its editing method and application. red9 A plasmid containing the gene and sgRNA elements is constructed, and specific primers are designed for the editing site to obtain the pTarget plasmid; the donor DNA sequence is constructed into a vector to obtain the pEdit plasmid. After phage is infected with a host bacteriophage containing the pEdit plasmid, the pTarget plasmid is used to reverse screen for mutant phage, achieving efficient and rapid gene editing of the phage genome, including gene deletion mutations, single nucleotide substitution mutations, and insertion mutations. This method is simple to operate and has high gene editing efficiency. It has promoted the development of structural analysis and functional research of Staphylococcus aureus phage genomes and provided a new strategy for the modification of engineered phages, with broad application prospects and market value.

The invention belongs to the technical field of microorganisms and genetic engineering. The invention provides a listeria monocytogenes strain as well as a preparation method and application thereof, the flagellum formation ability of the strain is enhanced, the environmental stress resistance is enhanced, and the immune recognition of host cells on listeria monocytogenes can be enhanced; the strain is subjected to delta yxeA gene deletion mutation; the bacterial strain is named as Listeria monocytogenes Lm-delta lmo2568, the preservation number of the bacterial strain is CGMCC (China General Microbiological Culture Collection Center) No.36001, and the bacterial strain is preserved in the China General Microbiological Culture Collection Center on September 22, 2025. The invention finds that the deletion of Listeria monocytogenes YxeA promotes the formation and expression of flagellum, and the obtained strain can enhance the immune recognition of host cells on Listeria monocytogenes, and can be used for preparing related preparations.

The invention discloses application of a rice OsNAR1 gene or a protein coded by the rice OsNAR1 gene to regulation and control of resistance of rice to pyricularia oryzae, and relates to the technical field of prevention and control of pyricularia oryzae. The CDS nucleotide sequence of the rice OsNAR1 gene is as shown in SEQ ID No. 2. Research finds that the rice gene OsNAR1 plays an important role in the rice blast resistance process of rice, the number of disease spots on leaves of a gene deletion mutant is smaller, the gene can be used for screening to obtain rice blast bacteria resistant rice strains, the expression quantity of basic resistance marker genes in the gene deletion mutant is remarkably increased, and the expression quantity of the basic resistance marker genes in the gene deletion mutant is remarkably increased. The expression of the basic resistance gene is negatively regulated by the OsNAR1, and the rice blast can be prevented and controlled.

The invention relates to a preparation method of PPAR gamma gene deleted zebrafish, belongs to the field of molecular biology, and particularly relates to preparation of PPAR gamma gene deleted mutant zebrafish. The method comprises the following steps: 1, synthesizing two sgRNA chains; 2, combining the two sgRNA chains with Cas9 protein, and microinjecting the combined sgRNA chains and Cas9 protein into fertilized eggs in a single cell period of zebra fish; 3, detecting mutation efficiency, and feeding the F0 generation zebrafish to adult fish; 4, backcrossing the F0-generation adult zebrafish and the wild type zebrafish; 5, selecting individuals with consistent filial generation deletion positions after backcross for selfing to obtain F1-generation PPAR gamma mutation zebrafish; and 6, carrying out selfing on the obtained F1 generation PPAR gamma deletion homozygous individuals. According to the invention, a PPAR gamma gene deletion mutant strain is obtained in zebrafish by using a gene editing technology, the designed double-acting target primer can realize accurate and efficient knockout of a target gene in a zebrafish genome, the knockout rate reaches 85%, and embryo development after microinjection does not have malformation. The obtained PPAR gamma gene deletion mutant zebrafish can be used for research on lipid metabolism and deposition.

The present invention provides a drought-resistant gene TaHsfC3-4 and its application in plant drought resistance improvement, relating to the field of plant genetic engineering technology. The drought-resistant gene TaHsfC3-4 of the present invention is cloned from the salt-alkali-tolerant / drought-resistant heat wheat variety Cang 6005, and the gene expression is induced by PEG6000 osmotic stress and ABA, and its nucleotide sequence is shown in SEQ ID NO.1, and the encoded amino acid sequence is shown in SEQ ID NO.2. The present invention utilizes Agrobacterium genetic transformation method to overexpress the gene in Arabidopsis wild type and Arabidopsis mutant (ABA receptor gene deletion mutant), and the drought resistance of the obtained transgenic strain is significantly improved, indicating that the TaHsfC3-4 gene cloned by the present invention has the function of improving plant drought resistance.

The invention discloses an LPLa gene deleted zebra fish primer and a preparation method of zebra fish, belongs to the technical field of molecular biology, and particularly relates to design and application of the LPLa gene deleted zebra fish primer. The LPLa gene deleted zebrafish primer disclosed by the invention consists of a forward primer and a reverse primer of two sgRNA (small guide ribonucleic acid); the method comprises the following steps: 1, synthesizing two sgRNA chains; 2, combining the two sgRNA chains with Cas9 protein, and carrying out micro-injection on the combined sgRNA chains and Cas9 protein into fertilized eggs in a single cell period of zebra fish; 3, detecting sgRNA knockout efficiency, and sequencing and screening mutant zebrafish F0; 4, backcrossing to obtain a mutant BC1 generation, and sequencing to detect the frame shift condition after the gene is knocked out; and 5, selfing to obtain F1-generation zebrafish LPLa- / -, and subsequently culturing to obtain the LPL gene deletion homozygous mutation line. The primer disclosed by the invention realizes accurate and efficient knockout of a target gene in a zebrafish genome, and the embryo development after microinjection does not have malformation, so that the LPLa gene deleted zebrafish mutant strain is obtained. The LPLa gene deletion mutant zebrafish provides a basic model for fish fat development and deposition regulation.

The present invention relates to an Arabidopsis thaliana MAP4K9 gene and its application in regulating the high-temperature tolerance of plants, belonging to the technical field of plant gene breeding. This gene has a nucleotide sequence as shown in SEQ ID NO.1, and the encoded protein of this gene has an amino acid sequence as shown in SEQ ID NO.2. By analyzing the MAP4K9 gene deletion mutant and the MAP4K9 overexpression transgenic Arabidopsis thaliana plants, the present invention finds that the MAP4K9 gene has a positive regulatory effect on the high-temperature tolerance of Arabidopsis thaliana plants, providing important theoretical significance and application value for regulating the high-temperature tolerance of plants.