61 results about "Gene deletion" patented technology

The invention relates to the technical field of genetic engineering, and provides a method for improving the yield of lincomycin by modifying a streptomyces lincomycin SLCG6736 gene, the SLCG6736 gene in the streptomyces lincomycin is deleted through a genetic engineering way to obtain a lincomycin high-yield engineering strain, and the obtained strain is used for fermentation production of lincomycin. Wherein the nucleotide sequence of the SLCG6736 gene is as shown in SEQ ID NO. 1. The invention further provides application of the method. The lincomycin biosynthesis negative regulator SLCG6736 has the advantages that the lincomycin biosynthesis negative regulator SLCG6736 is screened in the research, the SLCG6736 gene is knocked out through a genetic engineering way, a lincomycin high-yield strain can be obtained, and a technical support is provided for improving the lincomycin fermentation yield in industrial production.

The invention provides a method for improving the yield of lincomycin by modifying a streptomyces lincomycin SLCG3904 gene, which comprises the following steps: deleting a GlnR family transcriptional regulation gene SLCG3904 in the streptomyces lincomycin through a genetic engineering way to obtain a lincomycin high-yield engineering strain, and fermenting the obtained strain to produce the lincomycin. Wherein the nucleotide sequence of the SLCG3904 gene is as shown in SEQ ID NO. 1. The invention further provides application of the method. The invention has the advantages that the lincomycin biosynthesis negative regulator SLCG3904 (GlnR family) is screened in the research, the SLCG3904 gene on the streptomyces lincomycin chromosome is deleted through a genetic engineering way, a lincomycin high-yield strain can be obtained, and a technical support is provided for improving the lincomycin fermentation yield in industrial production.

The invention belongs to the field of plant disease control research, and relates to a cucumber corynespora cassiicola effect protein CcSP1 as well as a coding gene and application thereof. The invention provides a novel cucumber corynespora cassiicola effect protein CcSP1 as well as a coding gene and application thereof. The nucleotide sequence of the coding gene of the corynespora cassiicola effect protein CcSP1 is as shown in SEQ ID NO. 1. Through transient expression of the CcSP1 gene in nicotiana benthamiana, allergic necrosis caused by nicotiana benthamiana can be inhibited. A gene deletion mutant delta CcSP1 is constructed through homologous recombination, vegetative growth is not affected by deletion of the gene, but the pathogenicity is remarkably reduced when cucumber leaves are infected. The result shows that CcSP1 participates in scab expansion after plants are infected by corynespora cassiicola, and plays an important role in the pathogenic process of corynespora cassiicola. The invention has important significance for analyzing the related molecular mechanism of the cucumber corynespora leaf spot and establishing the comprehensive prevention and control technical strategy of the plant cucumber corynespora leaf spot.

The invention discloses an application of an FgGCV1 gene and a knockout mutant thereof in prevention and control of fusarium graminearum, and belongs to the technical field of microorganisms. The invention discloses for the first time that after the FgGCV1 gene is knocked out, generation of ascospores in the sexual reproduction process of fusarium graminearum can be inhibited, the fusarium graminearum is prevented and controlled, wheat scab is prevented and controlled, and the nucleotide sequence of the FgGCV1 gene is as shown in SEQ ID NO.1. The FgGCV1 gene deletion mutant specifically regulates and controls maturation of ascospores in the sexual reproduction process of fusarium graminearum, and the ascospores serve as an important primary infection source of wheat scab, so that the transmission path of the wheat scab can be blocked after the FgGCV1 gene is knocked out, and the FgGCV1 gene deletion mutant plays an important role in fusarium graminearum prevention and control and green ecological prevention and control; the method is also an important resource for researching the sexual reproduction process of filamentous ascomycetes.

The invention relates to application of a corn ZmECT3 gene in regulation and control of plant heat resistance, and belongs to the technical field of plant gene breeding, the nucleotide sequence of the ZmECT3 gene is as shown in SEQ ID NO.1, and the amino acid sequence of the gene encoding protein is as shown in SEQ ID NO.2. By analyzing a ZmECT3 gene deleted corn plant and a ZmECT3 gene overexpressed arabidopsis thaliana plant, it is found that the ZmECT3 gene can regulate and control the heat resistance of the plants, and important theoretical significance and application value are provided for creation of heat-resistant corn new germplasm and heat-resistant corn molecular breeding.

The invention discloses a method for preparing bovine MSTN gene editing cells by using CRISPR / Cas12i, and belongs to the technical field of biology. According to the invention, an sgRNA expression vector aiming at the bovine MSTN gene is constructed, and a CRISPR / Cas12i system is utilized to carry out precise gene editing on a specific target area of a first exon of the bovine primary fibroblast MSTN gene, so that mRNA of the MSTN gene cannot be normally transcribed, and MSTN gene deleted cells are obtained. Compared with other gene editing tools, the Cas12i system shows higher specificity and editing efficiency (improved by 3%-5%), and the off-target effect risk is effectively reduced. The prepared MSTN gene deleted bovine recombinant cell line is stable and efficient in editing efficiency, the cell expression level is not affected, the MSTN gene deleted bovine recombinant cell line can be widely applied to somatic cell cloning and microinjection preparation of double-muscle-hip beef cattle, and a new strategy is provided for beef cattle breeding.

The invention discloses a dual fluorescent quantitative PCR (polymerase chain reaction) primer probe combination for identifying a PRV (pseudorabies virus) vaccine strain and a wild strain. The dual fluorescent quantitative PCR primer probe combination comprises two pairs of primers gB-F and gB-R, gE-F and gE-R, and probes gB-probe and gE-probe, wherein the two pairs of primers are used for specifically amplifying a PRV gB gene and a PRV gE gene; through existence of real-time fluorescence PCR amplification curves of the PRV gB gene and the gE gene, a PRV vaccine strain (gE gene deletion) and a wild strain can be rapidly, specifically, sensitively and stably identified, technical support can be provided for porcine pseudorabies epidemic disease monitoring, and thorough purification of the PRV in a live pig breeding link is facilitated.

The invention discloses a fusarium venetianum engineering strain as well as a construction method and application thereof, relates to the technical field of gene engineering, and aims to solve the problem of low fungal hypha protein yield, chitin synthetase FvChs4 gene of fusarium venetianum is deleted, the classification name of the fusarium venetianum engineering strain is fusarium venetianum, and the fusarium venetianum engineering strain has the characteristics that the fusarium venetianum engineering strain has a nucleotide sequence shown in SEQ ID NO: 1 and a nucleotide sequence shown in SEQ ID NO: 2 and a nucleotide sequence shown in SEQ ID NO: 3. The fusarium venanii engineering strain is preserved in the China General Microbiological Culture Collection Center (CGMCC) on May 27, 2025, and the preservation number is CGMCC No. 42015. Through targeted knockout of the FvChs4 gene, consumption of a carbon source by cell wall synthesis is reduced, and directional enrichment of the carbon source to protein synthesis is realized, so that the yield of hypha protein is increased.

The application discloses a Toxoplasma gondii DAHPS gene deletion vaccine strain and application thereof. The application provides a Toxoplasma gondii gene knockout strain, which is obtained by directly knocking out a 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase TgDAHPS gene of the Toxoplasma gondii through a gene editing technology, the gene ID of the TgDAHPS gene is TGGT1_221260, and the nucleotide sequence is shown as SEQ ID NO. 1. The gene deletion strain Δdahps provided by the application can grow normally in vitro, does not reproduce in a host in vivo, and is almost non-toxic. After immunization of the Δdahps strain, the host can be induced to have good immune protection. The Δdahps has the advantages of weak virulence and almost no reproduction in vivo as the attenuated vaccine strain of the Toxoplasma gondii, can improve the resistance of the host to wild Toxoplasma gondii, and can be used for preventing the infection of the Toxoplasma gondii in humans and animals, and has important significance for developing more Toxoplasma gondii vaccines.

The invention discloses a recombinant vector for expressing varIBDV VP2 gene Meg Gene-deficient Marek's disease virus strain and its construction method and application. Meg The gene-deficient Marek's disease virus strain is obtained by using recombinant cloning technology to insert the gene fragment CMV-VP2 containing the CMV promoter sequence, varIBDV VP2 gene and terminator sequence into the Meg The gene-deleted Marek's disease virus strain rMSΔMeq US2 gene was obtained by replacing the nucleotide sequence between positions 156501-157116 of the rMSΔMeq genome, and was named rMDV-varVP2. Studies have shown that the recombinant obtained by the present invention Meg The gene-deficient Marek's disease virus rMDV-varVP2 has the same in vitro replication ability and good genetic stability as the parent virus rMSΔMeq strain, and rMDV-varVP2 can provide good protection against IBDV mutants after immunizing chickens. The present invention provides a technical means for the development of a recombinant MDV live vector vaccine.

The invention provides a novel recombinant orf virus, the recombinant orf virus expresses interleukin 15, the recombinant orf virus is an ORFV (Open Reading Frame Virus) ORFs 005-008 gene deleted strain (marked as ORFV delta), and the IL15 gene expressed by the recombinant orf virus is one of a mouse source, a human source, a cat source or a dog source. According to the invention, an ORFV oncolytic virus is used as a carrier to develop a tumor immunotherapy drug ORFVdelta-IL15 expressing IL15, and the IL15 is expressed in tumor cells by using the characteristics of virus infection, so that the activation of NK cells and T cells in a tumor microenvironment is promoted while the tumor cells are cracked, and the anti-tumor effect of the oncolytic virus is enhanced. The invention provides a new strategy for tumor treatment.

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 invention provides a method for preventing or treating diseases related to CSNK2B gene deletion. The disclosure also provides a CSNK2B protein or a variant thereof, a nucleic acid molecule encoding a CSNK2B protein or a variant thereof, an expression cassette comprising a nucleic acid molecule encoding a CSNK2B protein or a variant thereof, a vector comprising a nucleic acid molecule encoding a CSNK2B protein or a variant thereof, a virus, such as an AAV particle, expressing a CSNK2B protein or a variant thereof, and a method for preparing the CSNK2B protein or a variant thereof. According to the present invention, the use of the cells expressing the CSNK2B protein in the preparation of drugs for treating or preventing CSNK2B gene deletion-related diseases in subjects is provided, and the CSNK2B gene deletion-related diseases are selected from the group consisting of Rett syndrome, autism spectrum disorder (ASD), Porier-Bienvenu neurodevelopment syndrome (POBINDS), epilepsy or broad-spectrum development retardation, or the use of the cells expressing the CSNK2B protein in the preparation of the drugs for treating or preventing the CSNK2B gene deletion-related diseases in the subjects,

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 invention discloses a toxoplasma gondii gene deletion strain ME49 [delta] TGZS and an application thereof in preparation of a gene engineering inactivated vaccine. The toxoplasma gondii gene deletion strain ME49 [delta] TGZS is prepared by taking a toxoplasma gondii strain ME49 as a parent strain and knocking out a TGZS gene in the toxoplasma gondii strain ME49; the nucleotide sequence of the TGZS gene is as shown in SEQ ID NO: 1. The strain can maintain normal polypide growth and replication capacity under an in-vitro culture condition, after inactivation treatment, the strain loses proliferative activity but maintains a complete polypide morphological structure, and the strain is compatible with an aluminum hydroxide adjuvant to prepare a vaccine preparation. According to the vaccine, by activating the specific immune response of a terminal host cat, the sexual reproduction cycle of the toxoplasma gondii in the intestinal epithelial cells of the cat is effectively blocked, and the formation and discharge of oocysts are remarkably inhibited, so that the propagation chain of the toxoplasma gondii is controlled from the source. The invention provides a new effective means for prevention and control of toxoplasmosis.

The invention discloses a Brucella bacterium capable of generating a lipoid A vaccine adjuvant and an application of the Brucella bacterium. According to the invention, recombinant plasmids for expressing BacA and LpxL genes are constructed, and are transformed into Brucella through electric shock, so that a double-gene knockout strain is successfully obtained. By knocking out BacA, an ultra-long fatty acid chain of lipoid A is shortened, an organism is induced to generate an inflammatory reaction, and by knocking out LpxL, the inflammatory reaction is reduced, so that the effect of the immunologic adjuvant is achieved. LPS is extracted for in-vitro stimulation analysis, and it is found that the inflammatory response of double-gene deletion is reduced compared with that of single-gene deletion. Animal experiments show that double-gene deletion can induce an organism to generate good immune response and induce relatively high cellular immunity and humoral immunity, and a new thought is provided for research and development of gram-negative bacterium inactivated vaccines.

The invention provides a method for improving the yield of thaxtomin A through a streptomyces scabies SCAB82261 gene. The method comprises the following steps: deleting the streptomyces scabies SCAB82261 gene through genetic engineering to obtain a thaxtomin A high-yield strain, and carrying out fermentation production; wherein the nucleotide sequence of the SCAB82261 gene is as shown in SEQ ID NO. 1, and the coded amino acid sequence is as shown in SEQ ID NO. 2. The invention has the following advantages: the thaxtomin A biosynthesis negative regulation gene SCAB82261 is screened, and the PaaK protein encoded by the thaxtomin A biosynthesis negative regulation gene SCAB82261 can catalyze the degradation of aromatic compounds such as phenylalanine; when the streptomyces scabies SCAB82261 gene is deleted, a high-yield strain of the thaxtomin A can be obtained, and a technical support is provided for improving the yield of industrial production of the thaxtomin A.

This invention discloses a method for rice to withstand heat stress. RMI1 Genes and their applications. Among them... RMI1 The nucleotide sequence of the gene is shown in SEQ ID No. 1, and the amino acid sequence of the protein it encodes is shown in SEQ ID No. 2; this invention verifies RMI1 The function of the gene, RMI1 protein is located in the cell nucleus, rmi1-1 In mutants RMI1 The deletion of 33 base pairs in the gene results in the loss of 11 amino acids in the RMI1-1 protein, making... rmi1-1 High temperatures during the seedling stage cause a large number of abnormal chromosomes to adhere and fragment during the late mitotic phase of root tip cells, resulting in stunted plant growth and development. rmi1-1 High temperatures during the reproductive growth period of plants lead to a large number of chromosome fragments and adhesions in the late stage of meiosis I, resulting in sterility; high temperatures also affect the interaction between RMI1-1 protein and topoisomerase TOP3α and helicase RECQ4. rmi1-cr3 Frameshift mutations lead to abnormal embryo and endosperm development; overexpression RMI1 This invention can enhance the heat resistance of rice. RMI1 The analysis of gene function provides genetic resources for the breeding of heat-resistant rice varieties.

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 invention discloses a toxoplasma gondii DAHPS gene deletion vaccine strain and an application thereof. The invention provides a toxoplasma gondii gene knockout strain, the toxoplasma gondii gene knockout strain is obtained by directly knocking out a 3-deoxy-D-arabinoheptulose-7-phosphate synthase TgDAHPS gene of toxoplasma gondii through a gene editing technology, the gene ID of the TgDAHPS gene is TGGT1221260, and the nucleotide sequence of the TgDAHPS gene is shown as SEQ ID NO.1. The toxoplasma gondii gene knockout strain has the advantages that the toxoplasma gondii gene knockout strain can be used for preparing the toxoplasma The gene-deleted insect strain delta dahps provided by the invention can normally grow in vitro, does not propagate in a host and is almost non-toxic, and after the delta dahps insect strain is immunized, the host can be induced to generate good immune protection force. As a toxoplasma gondii attenuated vaccine strain, the delta dahps has the advantages of weak toxicity and almost no reproduction in vivo, can improve the resistance of a host to wild toxoplasma gondii, can be used for preventing human and animals from being infected with toxoplasma gondii, and has important significance for developing more toxoplasma gondii vaccines.

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 application relates to application of a maize ZmECT3 gene in regulating heat tolerance of plants, belongs to the technical field of plant gene breeding, and discloses that the nucleotide sequence of the ZmECT3 gene is shown as SEQ ID NO. 1, and the amino acid sequence of a protein coded by the gene is shown as SEQ ID NO. 2. Through analysis of maize plants with a ZmECT3 gene deletion and Arabidopsis plants with overexpression of the ZmECT3 gene, it is found that the ZmECT3 gene can regulate heat tolerance of plants, and the application provides important theoretical significance and application value for creating new heat-tolerant maize germplasm and heat-tolerant maize molecular breeding.

The invention discloses a highly toxic Klebsiella pneumoniae rpoE The invention relates to a deletion strain and its application in preparing attenuated live vaccines, belonging to the field of microbiology. rpoE The killing effect of hvKP gene on wax moth was weakened, and the number of colonization in mouse viscera was reduced, and the damage to the lung was reduced, indicating that the knockout of high-virulence Klebsiella pneumoniae rpoE The gene can reduce the pathogenicity of highly virulent Klebsiella pneumoniae. At the same time, immune protection experiments have shown that highly virulent Klebsiella pneumoniae rpoE The attenuated live vaccine prepared from the gene-deleted strain provides high immune protection for mice, and has better immune protection than attenuated killed vaccines, with broad market application prospects. This invention broadens the research on the pathogenesis of hvKP and provides a theoretical basis for the prevention and treatment of hvKP.

The invention discloses a method for distinguishing a vibrio alginolyticus wild type virulent strain and an ApA gene deletion type weak strain based on lipidomics. The ratio of the content of CL (16: 0 / 16: 1 / 16: 1 / 16: 1 / 16: 1) to the content of LPE (20: 0) in vibrio alginolyticus is used for distinguishing a vibrio alginolyticus sRNA molecule AphA virulent strain and a vibrio alginolyticus attenuated strain in a lipidomics research, the vibrio alginolyticus sRNA molecule AphA virulent strain with the ratio of CL to LPE being more than 15 in a sample to be detected is a vibrio alginolyticus wild type virulent strain, and the vibrio alginolyticus AphA gene deleted attenuated strain with the ratio of CL to LPE being less than 1 in the sample to be detected is a vibrio alginolyticus AphA gene deleted The method disclosed by the invention is simple to operate and reliable in detection result, and can provide reference for lipid characteristic analysis of the vibrio alginolyticus sRNA molecule AphA and related research on lipidomics of the vibrio alginolyticus sRNA molecule AphA.

The invention discloses a mycoplasma bovis protein RecD as well as a coding gene, a recombinant vector and application thereof, and aims to solve the technical problems that safe and effective commercial mycoplasma bovis vaccines are lacked in the current market and the function research on mycoplasma bovis genes is insufficient. A recombinant protein RecD is successfully expressed by optimizing a sequence of a mycoplasma bovis gene Mbov0848, constructing a recombinant vector and transforming the recombinant vector into escherichia coli, the recombinant protein RecD is confirmed to have ATP enzyme activity through purification and functional verification, the optimal reaction conditions of the recombinant protein RecD are further determined, meanwhile, an Mbov0848 gene deleted strain is constructed, and the recombinant protein RecD is transformed into escherichia coli. It is found that the viability of a strain without the gene in a host is reduced. The application has important significance for understanding the pathogenic mechanism of the diseased mycoplasma bovis and developing a new treatment strategy and a new mycoplasma bovis vaccine.

The invention discloses a toxoplasma gondii CFAP gene deleted strain as well as a construction method and application thereof. The toxoplasma gondii CFAP gene deleted strain ME49 cfu is prepared by taking a toxoplasma gondii strain ME49 as a parent strain and knocking out a CFAP gene in the toxoplasma gondii strain ME49; the nucleotide sequence of the CFAP gene is as shown in SEQ ID NO: 1. The strain ME49 cfap can be normally cultured in vitro, the polypide can normally grow and replicate, but the strain can induce the terminal host cat to generate immune protection force, inhibit the sexual reproduction of the toxoplasma gondii, reduce the oocyst discharge, and facilitate the reduction of the transmission risk of the toxoplasma gondii caused by the terminal host cat.

The invention relates to a novel oncolytic vaccinia virus vector VVL [delta] TK [delta] N1L [delta] A41L-RFP and VVL [delta] TK [delta] N1L [delta] A41L-IL-27 with deletion of A41L genes and an application of the novel oncolytic vaccinia virus vector VVL [delta] TK [delta] N1L [delta] A41L-IL-27. The novel oncolytic vaccinia virus vector VVL [delta] TK [delta] N1L [delta] A41L-RFP is a vaccinia virus Lister strain, a TK gene, an N1L gene and an A41L gene are deleted at a fixed point, and the vector has tumor targeting, can carry an exogenous therapeutic gene, and is used for treating various solid tumors. The vaccinia virus vector VVL [delta] TK [delta] N1L [delta] A41L-IL-27 can kill tumor cells and express interleukin 27 at the same time, and the vaccinia virus vector VVL [delta] TK [delta] N1L [delta] A41L-IL-27 and the oncolytic vaccinia virus have a stronger anti-tumor effect. The virus vectors VVL [delta] TK [delta] N1L [delta] A41L-RFP and VVL [delta] TK [delta] N1L [delta] A41L-IL-27 provided by the invention can be used as a potential novel gene therapy drug for treating pancreatic cancer, colon cancer, lung cancer and other tumors.

The present invention discloses a guide RNA for constructing a gene-deficient Arabidopsis mutant plant, an Arabidopsis mutant plant and a preparation method thereof, wherein the guide RNA comprises a pair of sequences as shown in SEQ ID No: 1 and SEQ ID No: 2. The preparation method comprises: designing a primer pair according to the guide RNA; using the primer pair for PCR amplification, purification, and enzyme digestion, and then connecting with a vector to obtain a recombinant plasmid; using the recombinant plasmid to transform competent cells, and then screening and verifying to obtain an expression vector for knocking out a gene to be knocked out in Arabidopsis; transforming the obtained expression vector into Agrobacterium and soaking the flowers, and then screening to obtain an Arabidopsis mutant plant. The technical solution of the present invention overcomes the current shortcomings in plant breeding and other performances, can advance the bolting period of Arabidopsis, and provides important value for Arabidopsis in the direction of plant breeding.

The invention relates to the field of construction of gene deletion strains, in particular to a Brucella M5bvfA gene deletion strain as well as a construction method and application thereof. The specific construction method comprises the following steps: connecting and synthesizing upstream and downstream homologous recombination sequences of a bvfA gene to obtain a targeting fragment, and cloning the targeting fragment into a suicide plasmid containing a gentamicin resistance gene to obtain a targeting plasmid; the targeting plasmids are electrically transformed into escherichia coli, and donor bacteria are obtained; the method comprises the following steps: jointing donor bacteria with brucella M5 recipient bacteria, and screening to obtain gentamicin-resistant brucella clones; the method comprises the following steps: inoculating gentamicin-resistant brucella cloned bacterial liquid into a culture medium containing cane sugar, culturing until monoclone is formed, and identifying and screening to obtain a traceless brucella M5bvfA gene deleted strain. The deletion strain can distinguish natural infection from vaccine immunity, has no antibiotic gene residue, and has the advantages of low toxicity and traceless deletion.

This invention discloses tomatoes WRKY52 The application of genes in regulating plant resistance to insect pest stress belongs to the field of biotechnology. WRKY52 The nucleotide sequence of the protein-coding region of the gene is shown in SEQ ID NO.1. This invention discloses tomato for the first time. WRKY52 The role of genes in regulating plant resistance to insect pests, especially bollworm, and the use of CRISPR / Cas9 gene editing technology to obtain tomatoes WRKY52 The study found that the mutant with gene deletion exhibited reduced resistance to the cotton bollworm, proving... WRKY52 The application of genes as positive regulators in improving the resistance of tomatoes to insect pest stress can be used for the breeding of insect-resistant tomato varieties. This invention provides more gene resources and new ideas for plant molecular stress resistance genetic breeding and genetic modification of plant stress tolerance.