48 results about "Deletion mutant" 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 quinoa CqARF2 gene mutant as well as an identification method and application thereof. Chenopodium quinoa seeds are mutated on a large scale by utilizing a chemical mutagenic agent EMS, a CqARF2 gene function deletion mutant arf2 is adopted as a quinoa mutant, G at the 1050th site of the CqARF2 gene of the mutant is mutated into A, and amino acid at the 350th site is mutated into a termination codon, so that the translation process of CqARF2 protein is terminated in advance. The chenopodium quinoa CqARF2 gene function deletion mutant arf2 shows remarkable agronomic character improvement, seeds are large and full, compared with a wild type, the hundred-grain weight of the mutant seeds is increased by about 40%, the seed diameter is increased by about 10%, the seed thickness is increased by about 30%, and a certain foundation is laid for breeding new high-yield and high-quality chenopodium quinoa varieties.

The invention belongs to the technical field of plant biology, and provides a method for regulating and controlling the heading stage of rice by using an OsTCP2 gene, and the OsTCP2 is used as a TCP family transcription factor and plays an important role in regulating and controlling the heading stage of the rice. Experimental results show that the heading period of the OsTCP2 function deletion mutant strain is obviously advanced. The nucleotide sequence of the OsTCP2 gene is defined, the mutant of the OsTCP2 gene is constructed, and an OsTCP2 mutant plant is successfully obtained through a CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-associated 9) technology. The mutants show an obvious early blossoming phenomenon under a long-day condition, and the heading period is about 8 days earlier than that of a wild type on average. A new gene resource is provided for regulation and control of the heading period of the rice, rice varieties can be improved through a molecular breeding means, the adaptive capacity and yield of the rice varieties in different regions are improved, and the gene has important significance on agricultural production.

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 application belongs to the field of new application of functional proteins and mutants thereof, and discloses application of sugarcane ScCBS protein and key domain deletion mutants thereof. The nucleotide sequence of the sugarcane ScCBS protein is shown as SEQ ID NO:1, and the amino acid sequence is shown as SEQ ID NO:2. The key domain deletion mutants of the ScCBS protein are ScCBS△motif1, ScCBS△motif3, ScCBS△motif4 and ScCBS△motif9, which respectively lack 28, 50, 49 and 29 amino acids. The application proves that the ScCBS protein can interact with SCSMV P1 and inhibit the RNA silencing suppressor activity by means of yeast two-hybrid, two-molecular fluorescence complementary experiment and agrobacterium transient expression system; the four mutants all lose the interaction and inhibition activity. Therefore, the ScCBS protein and the key domain deletion mutants thereof can be used for cultivating excellent SCSMV-resistant materials, and have wide application prospects in sugarcane disease-resistant molecular breeding.

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 belongs to the technical field of plant genetic engineering and molecular breeding, and particularly relates to a new function and application of a transcription factor gene OsMYB87 derived from rice, an OsMYB87 function deletion mutant created through a gene editing technology and application of the OsMYB87 function deletion mutant in increasing the rice grain length and grain weight. The OsMYB87 gene found in the invention shows a key differentiated characteristic: preliminary phenotypic observation shows that the phenomenon that chalkiness of endosperm is remarkably aggravated inevitably due to OsMYB73 knockout is not observed while the function deficiency of the OsMYB87 gene brings a remarkable yield increasing effect (grain length and grain weight increase). The OsMYB87 is strongly prompted to be possibly located on a different regulatory pathway, and as a breeding target, the OsMYB87 provides greater possibility and a better potential choice for maintaining the appearance quality of rice commodities while realizing yield increase, so that the OsMYB87 has higher application value.

The invention relates to a rice OsACC gene promoter mutant for resisting ACCase inhibitor herbicides and application of the rice OsACC gene promoter mutant. The rice OsACC gene mutant is a deletion mutant or a deletion and combination insertion mutant; the deletion mutation occurs in the promoter of the rice OsACC gene, and the sequence of the nucleic acid before deletion mutation is as shown in SEQ ID No. 1; the length of the sequence of the deleted nucleic acid is 29 bp to 97 bp, A in the initiation codon ATG of the OsACC gene is 0 site, the deleted nucleic acid occurs in the 305-209 sites of the upstream of the initiation codon ATG, and at least the 271-243 sites of the upstream of the initiation codon ATG are deleted; the inserted nucleic acid occurs within sites 271 to 243 upstream of the initiation codon ATG. The mutant can enable rice to obtain resistance to at least one of herbicides such as haloxyfop-R-methyl, fluazifop-p-butyl and clethodim.

The invention discloses a monoclonal antibody capable of simultaneously identifying mycobacterium tuberculosis secretory protein 64 of a wild type (W-MPT64) and a 63bp deletion mutant type (D-MPT64). The amino acid sequences of variable regions of a heavy chain and a light chain of the antibody are shown as SEQ ID NO.1 and SEQ ID NO.5. The lowest detection limits of W-MPT64 and D-MPT64 by a mycobacterium tuberculosis MPT64 double-antibody sandwich enzyme-linked immunosorbent assay and a fluorescence immunochromatography assay established by using the monoclonal antibody both reach 1.5625 pg / mL, which are significantly higher than the lowest detection limit in the prior art, and the monoclonal antibody can be used for qualitative and quantitative detection of mycobacterium tuberculosis secretory protein 64.

The invention discloses a vascular specific VND gene promoter and application of the vascular specific VND gene promoter in driving expression of xylan hydrolase in plants. The invention provides a DNA fragment. The DNA fragment is any one of the following: 1) the nucleotide sequence of the DNA fragment is sequence 1; and 2) a nucleic acid sequence which has more than 90% of homology with the nucleic acid sequence shown in 1) and has the same function. Experiments prove that a SbNAC31 gene promoter specifically expressed by a vascular tissue is connected with CDS of a xylan hydrolase gene SbXyl, and a vector with the fusion gene is used for converting a sorghum xylan hydrolase function deletion mutant, so that the growth and development defects of the mutant are made up; meanwhile, the excellent lignocellulose quality (high feed quality) of the mutant is reserved.

The present application belongs to the technical field of genetic engineering, and particularly relates to a genetically engineered strain with high yield of acetoin and 2,3-butanediol, a construction method and application. The present application obtains acoR deletion mutant MW03-ΔA by knocking out the regulatory gene acoR of the acetoin dehydrogenase enzyme system, blocks the degradation of AoDHES to acetoin, effectively reduces the production of by-products, thereby facilitating the accumulation of more acetoin. A strong promoter expression vector is further constructed in the MW03-ΔA strain, and the alpha-acetyl lactate decarboxylase gene alsD is connected, and then introduced into the acoR deletion mutant MW03-ΔA of Bacillus licheniformis, so as to overexpress alpha-acetyl lactate decarboxylase, and obtain a genetically engineered strain MW-BL4 with high yield of acetoin and 2,3-butanediol. The present application has the beneficial effect that compared with a single product of the engineered strain, the present application can realize the production of acetoin and 2,3-butanediol and other multiple products by using a single strain, and the yield is considerable, which can significantly reduce the cost of bacterial strains in industrial production and improve the production efficiency.

The invention relates to a herbicide-resistant rice OsACC gene promoter mutant and application thereof. The rice OsACC gene mutant is a deletion mutant; wherein the deletion mutation occurs in the promoter of the rice OsACC gene, and the sequence of the nucleic acid before the deletion mutation is shown as SEQ ID No.1; the length of the sequence of the deleted nucleic acid is 14 bp to 30 bp, A in the initiation codon ATG of the OsACC gene is 0 site, the deleted nucleic acid occurs in 964 to 935 sites on the upstream of the initiation codon ATG, and at least bases on 958 to 945 sites on the upstream of the initiation codon ATG are deleted. The mutant can enable rice to obtain resistance to herbicide fluazifop-p-butyl and / or clethodim.

The invention discloses application of a rice nitrogen utilization gene NU3 in regulation and control of rice growth, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the rice nitrogen utilization gene NU3 disclosed by the invention is as shown in SEQ ID NO. 1; the regulation of rice growth refers to regulation of rice nitrogen utilization rate, regulation of rice seed quality and regulation of seed protein content. According to the invention, the expression mode of the gene NU3 in rice is identified, and a function deletion mutant of the gene NU3 is obtained by using a gene editing technology. The rice seedling growth amount of the functional deletion mutant is reduced, the thousand seed weight is obviously reduced, the protein content of seeds is reduced, and the nitrogen utilization rate is reduced. The invention provides new gene resources and research basis for rice nitrogen metabolism research, growth regulation mechanism research and seed quality regulation.

Engineered Bacteroides thetaiotaomicron (Bt) strains, including deletion mutant strains, and methods of use thereof are provided. Methods of delivering a therapeutic compound to a gut microbiome of a subject include generating an engineered Bacteroides thetaiotaomicron (Bt) strain to overproduce outer membrane vesicles (OMVs); loading a target therapeutic compound into the OMVs; and colonizing the gut microbiome of the subject with the engineered Bt strain. In some embodiments, the subject has at least one of lactose intolerance, phenylketonuria, inflammatory bowel disease, and a chronic intestinal condition.

The invention relates to a herbicide-resistant rice OsACC gene mutant and application thereof. The rice OsACC gene mutant is a deletion mutant; the sequence of the nucleic acid before mutation is deleted is as shown in SEQ ID No. 1; the length of the sequence of the deleted nucleic acid is 33-97 bp, A in the initiation codon ATG of the OsACC gene is 0 site, the deleted nucleic acid occurs in the 305-209 sites of the upstream of the initiation codon ATG, and at least the 251-219 sites of the upstream of the initiation codon ATG are deleted; and excluding the following mutants: the length of the sequence of the deleted nucleic acid is 53-97 bp, the deleted nucleic acid occurs in the 305-209 sites upstream of the initiation codon ATG, and at least the 271-219 sites upstream of the initiation codon ATG are deleted. The mutant can enable rice to obtain three resistances of haloxyfop-R-methyl, fluazifop-p-butyl and clethodim.

The invention relates to a plant receptor-like kinase gene OsER1 participating in regulation and control of rice spikelet development, and compared with a wild type, an OsER1 gene function deletion mutant for regulating and controlling rice spikelet development and seed morphology has the advantage that the number of glumes, inner and outer hulls, stamens, pistils, stigmas and lobes of oser1-T is partially deviated. As the number of pistils in one spikelet is increased, the number of seeds is increased, and phenotypes of abnormal seed forms are avoided. The research proves that the OsER1 influences the pistil number and seed morphology of rice by regulating and controlling the development of rice glumous flowers. The discovery has important significance on creation of rice genetic materials and crop genetic improvement work, and has certain application value on rice breeding work.

One embodiment of the present disclosure is a composition for inhibiting the accumulation of α-synuclein, the composition containing an α-synuclein deletion mutant or a nucleic acid construct that expresses the deletion mutant.

This invention discloses a method for constructing a Brucella bovis mutant and its application in outer membrane vesicles. Using Brucella bovis RB51 as the parent strain, a triple-deletion mutant was obtained through multi-gene editing and homologous recombination by stepwise knockout of the eipB, per, and wadC genes. This invention, by knocking out genes such as eipB, constructs a mutant with significantly increased OMV secretion, smaller particle size, and superior immunoprotective efficacy compared to the parent strain. Furthermore, it is safe and lacks replication ability, laying the foundation for the development of a highly effective and safe brucellosis vaccine.

Provided is the use of an OsNPF7.2 gene in improving drought tolerance in rice breeding. A rice OsNPF7.2 gene deletion mutant is obtained by means of genetic engineering technology, thus confirming that the OsNPF7.2 gene can affect the drought tolerance of rice. An OsNPF7.2 gene overexpression line is obtained by means of genetic engineering technology, and a Japonica rice near-isogenic line carrying the IR64-type OsNPF7.2 gene is obtained by hybridization. Experimental results show that under drought conditions, both the OsNPF7.2 overexpression line and the Japonica rice near-isogenic line carrying the IR64-type OsNPF7.2 gene exhibit a higher tiller number per plant and a higher yield than those of the wild type, indicating that increasing the expression of the OsNPF7.2 gene can enhance the drought tolerance of rice, and the IR64-type OsNPF7.2 gene has a better effect on improving drought tolerance.

An immunogenic composition comprising a deletion mutant of a Clostridioides difficile TcdB toxin (such as TcdB2 or TcdB1) that lacks residues at least from amino acid residue 1769 to amino acid residue 1787 of a wild-type TcdB amino acid sequence or of a protein having high identity thereto, a vaccine comprising the immunogenic composition, a method of stimulating an immune response, a nucleic acid which encodes the amino acid sequence of the deletion mutant, a vector encoding the nucleic acid, and a host cell comprising the vector.

The present invention provides a recombinant vector for producing a fibroblast growth factor 2 (FGF-2) protein, the recombinant vector comprising a bSA: FGF-2 gene sequence region, the bSA: FGF-2 gene sequence region sequentially comprises genes of a bovine serum albumin (bSA) coding sequence, a His tag coding sequence, a protease recognition site coding sequence and a fibroblast growth factor 2 (FGF-2) protein coding sequence; the bovine serum albumin (bSA) coding sequence is a sequence for coding a region containing wild-type bovine serum albumin (bSA-f) or a deletion mutant of bovine serum albumin, and the deletion mutant of bovine serum albumin contains 110 or more N-terminal amino acid residues of bovine serum albumin.

The application discloses a Rhodococcus double oxygenase deletion library and a construction method and application thereof. Based on strict bioinformatics screening, the Rhodococcus double oxygenase gene is screened, different double oxygenase genes are knocked out, the Rhodococcus double oxygenase deletion mutant library is constructed, and the library is used for systematic degradation function screening and genotype-phenotype correlation analysis of aromatic compounds, so that key double oxygenase genes necessary or redundant for degradation of specific aromatic compounds can be rapidly identified, the substrate utilization spectrum and functional division spectrum of the double oxygenase family can be drawn, and the application prospect in multiple fields such as high value of lignin, synthesis of fine chemicals, and environmental biological remediation is potential.

This invention discloses a SNP marker and detection primer pair associated with the adventitious root primordium trait of *Senecio scandens* with stem nodules, and its application, belonging to the field of molecular biology technology. Based on SNP marker technology, this invention develops an SNP marker and detection primer pair linked to the adventitious root primordium trait of *Senecio scandens* with stem nodules. This invention involves phenotypic identification of mutants induced by EMS mutagenesis in *Senecio scandens* with stem nodules, followed by isolation, purification, population construction, and genetic analysis combined with Mutmap. + The SNP marker associated with adventitious root primordia is located at 10,433,968 bp on chromosome 4 of *Senecio scandens*, involving a C / T mutation. When the genotype is T, *Senecio scandens* lacks adventitious root primordia. This invention's SNP molecular marker can screen F2 populations at the early cotyledon stage, identifying mutants and wild types without phenotypic differences, thus improving screening efficiency, shortening the breeding cycle, and demonstrating value in marker-assisted breeding.

The invention relates to a rice OsACC gene promoter mutant resistant to fluazifop-p-butyl or clethodim and application of the rice OsACC gene promoter mutant resistant to fluazifop-p-butyl or clethodim. The rice OsACC gene mutant is a deletion mutant; the sequence of the nucleic acid before mutation is deleted is as shown in SEQ ID No. 1; the length of the sequence of the deleted nucleic acid is 38-97 bp, A in the initiation codon ATG of the OsACC gene is 0 site, the deleted nucleic acid occurs in the 305-209 sites of the upstream of the initiation codon ATG, and at least the basic groups of the 305-268 sites of the upstream of the initiation codon ATG are deleted; the length of the sequence of the deleted nucleic acid is 63-97 bp, the deleted nucleic acid occurs in the 305-209 sites upstream of the initiation codon ATG, and at least the bases at the 305-243 sites upstream of the initiation codon ATG are deleted.

The invention provides a construction method of a male sterile line of corn, and relates to the technical field of plant breeding, the method comprises the following steps: designing double sgRNAs on two sides of a key functional region of a nuclear fertility gene of corn, and inducing exon region deletion mutation through Cas9 mediated double-site cutting; introducing the editing element into a donor plant, hybridizing with a receptor selfing line, backcrossing to a BC2 generation, and selfing to obtain a homozygous deletion mutant without the editing element; the method comprises the following steps: constructing a multi-control maintenance breeding construct containing a fertility restoring gene, a pollen inhibition module and a selection marker, enabling the multi-control maintenance breeding construct to exist in a maintainer line in a semi-zygote state, limiting the formation of male gametes containing the construct through the pollen inhibition module, and realizing automatic sorting of sterile line seeds by utilizing the selection marker. According to the scheme, complete and stable sterile characters are ensured through double sgRNA induced large fragment deletion, a non-transgenic homozygous mutant is obtained by combining a backcross selfing strategy, meanwhile, efficient and pure breeding of sterile line seeds is achieved through a multi-control maintenance system, and the hybrid seed production efficiency and the seed purity are remarkably improved.

The invention discloses application of a zinc metalloproteinase CsMep gene in regulating pathogenicity of colletotrichum orbiculare, and belongs to the technical field of biology. The zinc metalloproteinase CsMep is identified, and the nucleotide sequence of the CsMep gene is shown as SEQ ID NO.1. The zinc metalloproteinase CsMep not only regulates the zinc ion steady state of colletotrichum orbiculare, but also enhances the toxicity by influencing the metabolism of aromatic amino acids. Through knockout of the CsMep gene in colletotrichum gloeosporioides and construction of deletion mutant colletotrichum gloeosporioides, strain morphological observation and pathogen infection experiments show that the pathogenicity of the deletion mutant colletotrichum gloeosporioides is obviously reduced, a new thought is provided for a metalloproteinase-targeted colletotrichum gloeosporioides prevention and control strategy, and a theoretical basis is provided for development of novel bactericides.

The application discloses application of protein Fobut in regulation of pathogenicity of banana fusarium wilt fungus. The application obtains a deletion mutant and a back-supplementation mutant of Fobut by constructing a knock-out plasmid and a back-supplementation plasmid of a gene (Fobut) coding protein Fobut of banana fusarium wilt fungus and transforming corresponding protoplasts respectively. It is found through observation of the phenotype of the deletion mutant and the back-supplementation mutant and pathogenicity analysis that the knock-out gene Fobut can obviously reduce the pathogenicity of banana fusarium wilt fungus, and the pathogenicity is restored after back-supplementation, which indicates that the protein Fobut is related to the pathogenicity of banana fusarium wilt fungus and can be used as a target for prevention and treatment of banana fusarium wilt. The application enriches a protein database related to the pathogenicity of banana fusarium wilt fungus and is beneficial to prevention and treatment of banana fusarium wilt.

The application discloses a kind of simultaneously identifying wild type (W-MPT64) and 63bp deletion mutant (D-MPT64) Mycobacterium tuberculosis secreted protein 64 monoclonal antibody.The antibody heavy chain and light chain variable region amino acid sequence as shown in SEQ ID NO.1 and SEQ ID NO.5.The Mycobacterium tuberculosis MPT64 double antibody sandwich enzyme-linked immunosorbent assay and fluorescence immunoassay method established using the monoclonal antibody reach 1.5625 pg / mL, significantly higher than the minimum detection limit of prior art, can be used for qualitative and quantitative detection of Mycobacterium tuberculosis secreted protein 64.

The invention provides a specific InDel molecular marker of a key gene OsSPO11-1 for rice apomixis control and application of the specific InDel molecular marker, and belongs to the technical field of crop molecular heredity. The invention provides a 9bp deletion mutant of a second exon of a rice fertility control gene OsSPO11-1 at the 162-170bp position in a CDS (Coding Sequence) sequence. The mutation causes the deletion of three key amino acids His-Arg-Tyr in the amino acid coding sequences from the 54th site to the 56th site. In fertility expression, the homozygous type of the mutant shows the characteristic of complete sterility, and the heterozygous type of the mutant keeps complete fertility. In order to accurately identify the mutation, the invention specifically develops a specific molecular marker, as shown in SEQ ID NO.7. The invention also provides a method for identifying the mutation. Through single PCR amplification, the marker can accurately distinguish a wild type, a heterozygous type and a mutant type, and the detection result is completely consistent with the results of sequencing analysis and phenotype observation.

The invention relates to an E3 ubiquitin ligase coding gene and application thereof in resisting rice stripe virus. The method comprises the following steps: knocking out an OsRING18 gene from wild type ZH11 rice by utilizing a CRISPR / Cas9 gene editing system, so as to obtain a homozygous OsRING18 function deletion mutant rice strain; a result shows that compared with wild Nipponbare ZH11 rice, a function deletion mutant rice strain of the OsRING18 gene shows higher disease resistance.