127 results about "Promoter activity" patented technology

The invention discloses a tea tree CsU3 gene promoter and application thereof, and belongs to the technical field of plant biotechnology and gene engineering. Tea tree RNA polymerase III type promoters-tea tree endogenous U3 gene promoters CsU3a, CsU3b, CsU3c and CsU3d are successfully cloned for the first time, a tea tree U3 gene promoter activity detection carrier is successfully constructed, the transcriptional activities of the four CsU3 promoters are compared through instantaneous tobacco leaf conversion, GUS dyeing and GUS enzyme activity determination, and the CsU3 gene promoter activity detection carrier can be used for detecting the activity of the tea tree U3 gene promoter. CsU3a and CsU6b promoters with strong transcriptional activity are selected to be used for constructing a tea tree gene editing carrier, the feasibility of the promoter applied to a tea tree CRISPR / Cas9 gene editing system is verified in a tea tree protoplast, and CRISPR / Cas9 mediated tea tree genome targeted editing is realized.

The invention discloses haplotype molecular markers related to wheat drought resistance, primers, a kit and application of the haplotype molecular markers, and aims to solve the technical problem that molecular markers capable of identifying excellent haplotypes of TaXTH-2B gene promoters are lacked at present. It is confirmed that the TaXTH-2B gene promoter has two haplotypes, and the Hap II haplotype is an excellent haplotype endowing wheat with a high drought resistance index and has high promoter activity; meanwhile, the invention develops molecular marker primers for distinguishing the two haplotypes, and the molecular marker primers have wide application prospects in the aspects of identifying and screening bred wheat varieties or breeding intermediate generation materials, assisting in selecting wheat materials with high drought resistance indexes and the like.

The present invention relates to a nucleic acid sequence derived from a regulatory region of a human Lgr5 gene and having promoter activity in retina non-long-process cells. The invention also relates to an expression cassette or vector comprising the promoter, a viral particle or cell comprising the expression cassette or vector, and a pharmaceutical composition comprising the expression cassette or vector and the viral particle or cell. The invention also relates to the use of the expression cassette, vector, viral particle, cell or pharmaceutical composition in the treatment of eye diseases, especially diseases associated with retinal ganglion cell degeneration.

The invention provides a tobacco polyphenol oxidase gene NtPPO15 and application of the tobacco polyphenol oxidase gene NtPPO15 in inhibiting browning of tobacco and improving ripening resistance and baking resistance of the tobacco. According to the invention, a key gene NtPPO15 related to tobacco browning and baking resistance is screened through experiments, the activity of an NtPPO15 promoter and a gene expression site are further explored by constructing a GUS dyeing vector, and the function of the gene is deeply studied through gene editing and overexpression. Experiments prove that the NtPPO15 gene provided by the invention is a key gene related to browning in a tobacco maturing and baking process, and by regulating and controlling the expression quantity of the NtPPO15 gene, the browning of the tobacco can be controlled, the maturing resistance and baking resistance of the tobacco can be improved, the quality of a tobacco product can be improved, the harvesting frequency can be reduced, and the economic benefit can be greatly improved; and the method can also be used for screening tobacco varieties with excellent quality, so that the method has a good and wide market application prospect.

The present disclosure provides a method for producing a polyisoprenoid, which makes it possible to synthesize in vitro a polyisoprenoid having an unprecedented structure, such as a 100% cis-polyisoprenoid or a polyisoprenoid containing an allylic diphosphate derivative as an initiating terminal. The present disclosure relates to a method for producing a polyisoprenoid in vitro, which employs a gene coding for a neryl diphosphate synthase and rubber particles bound to a protein encoded by the gene, or a method for producing a polyisoprenoid, which includes introducing into a plant a vector in which a gene coding for a neryl diphosphate synthase is linked to a promoter having a promoter activity that drives laticifer-specific gene expression to express a protein encoded by the gene specifically in laticifers.

The invention discloses application of a VvENO2 gene in regulation and control of plant fruit development, and belongs to the technical field of plant genetic engineering. In the early stage, hydrogen peroxide (H2O2) treatment can significantly promote maturation of grape fruits, and transcriptome analysis on the grape fruits treated by H2O2 discovers that the expression quantity of the VvENO2 gene is reduced. In order to further clarify the function of the VvENO2 gene in the plant fruit development process, a subcellular localization test and promoter activity analysis are carried out on the VvENO2 gene, it is proved that the VvENO2 gene is localized in a cell nucleus, a promoter of the gene can start GUS reporter protein, and the activity of the VvENO2 gene can be enhanced through ABA and H2O2 treatment. Further, transgenic arabidopsis thaliana and transgenic grapes are constructed, and phenotypic observation proves that the VvENO2 gene negatively regulates the maturation of plant fruits. The invention provides a new regulation and control site for breeding of early-maturing varieties of plants.

The invention provides an application of inhibiting or knocking out a tobacco polyphenol oxidase gene NtPPO12 in inhibiting tobacco browning. The important gene NtPPO12 related to tobacco browning is screened out through transcriptome sequencing and molecular biological experiments, the NtPPO12 promoter activity and the gene expression site are further verified, and the gene function is deeply studied through gene editing and overexpression. Molecular biology experiments prove that the NtPPO12 gene provided by the invention is a key gene related to browning in the tobacco maturing and baking process, and the browning degree of tobacco can be controlled by regulating the expression quantity of the NtPPO12 gene, so that the maturing resistance and baking resistance of tobacco are improved, the harvesting frequency is reduced, the baking characteristic is improved, and the yield of tobacco is improved. Therefore, the method has a wide market application prospect.

The present invention relates to a nucleic acid sequence derived from a regulatory region of a human sox4 gene and having promoter activity in retinal ganglion cells. The invention also relates to an expression cassette or vector comprising the promoter, a viral particle or cell comprising the expression cassette or vector, and a pharmaceutical composition comprising the expression cassette or vector and the viral particle or cell. The invention also relates to the use of the expression cassette, vector, viral particle, cell or pharmaceutical composition in the treatment of eye diseases, especially eye diseases associated with retinal ganglion cell or photoreceptor cell degeneration.

The present application provides an application of inhibiting or knocking out tobacco polyphenol oxidase gene NtPPO12 in inhibiting tobacco browning. The present application screens an important gene NtPPO12 related to tobacco browning through transcriptome sequencing and molecular biology experiments, further verifies the promoter activity and gene expression site of NtPPO12, and deeply researches the function of the gene through gene editing and overexpression. The present application proves by molecular biology experiments that the NtPPO12 gene provided by the present application is a key gene related to browning in the mature curing process of tobacco, and by regulating the expression amount of the NtPPO12 gene, the browning degree of tobacco can be controlled, and the tobacco maturity and curing resistance can be improved, which is beneficial to reducing the picking frequency, improving the curing characteristics, and improving the appearance and internal quality of tobacco leaves, so it has a broad market application prospect.

The invention belongs to the technical field of genetic engineering, and particularly relates to a citrus canker pathogen mediated citrus leaf transient expression method which comprises the following steps: preparing a citrus canker pathogen infection solution which is xanthomonas citrus subspecies; infecting the citrus by using the citrus canker pathogen infection liquid, and culturing to obtain citrus infected by the citrus canker pathogen; preparing an agrobacterium infection solution containing a target gene; and inoculating the citrus infected by the citrus canker pathogen with an agrobacterium infection solution containing a target gene, and carrying out transient expression of the target gene. The transient expression method provided by the invention can realize transient and efficient expression of the target gene in leaves of different citrus varieties. The transient expression method disclosed by the invention can be used for researching gene functions, protein interaction, promoter activity analysis and the like; and the method can also be used for evaluating the citrus canker resistance gene function.

The invention discloses a porcine beta-defensin-3 gene core promoter as well as a construction method and application thereof, and belongs to the technical field of gene engineering. The sequence of the core promoter is as shown in SEQ ID No.1, and the core promoter has remarkable promoter activity in porcine small intestine epithelial cells and can be regulated and controlled by nutrient substances sodium butyrate and glutamine. The invention also discloses a construction method for specifically amplifying the core promoter, which comprises the following steps: carrying out PCR (Polymerase Chain Reaction) amplification by taking porcine small intestine epithelial cell DNA (Deoxyribonucleic Acid) as a template, constructing a pMD-18T-pBD-3-P recombinant plasmid by taking upstream and downstream primers as shown in SEQ No.2-SEQ No.3, and amplifying by taking the recombinant plasmid as a template to obtain a core promoter fragment. The core promoter disclosed by the invention provides an excellent experimental system for researching a transcription regulation mechanism of the porcine beta-defensin-3 gene, the nutritional response characteristic of the core promoter provides a new platform for researching nutrition-immune interaction, and the core promoter has a wide application prospect in the field of healthy breeding of livestock and poultry.

The present disclosure provides a method for obtaining a promoter sequence modified so as to have a desired transcriptional activity. More specifically, provided is a method for obtaining a promoter sequence modified so as to have a desired transcription activity, the method comprising: preparing an original promoter sequence to be modified; generating a set of a plurality of modified promoter sequences that can be created by genome editing techniques on the basis of the promoter sequence; predicting, by a machine learning model, the transcription activity of each modified promoter sequence included in the set of modified promoter sequences generated; and selecting a modified promoter sequence predicted to have a desired active transcription performance.

A method for treating a mental disorder in a subject, comprising the step of normalizing the expression of both DRD1 and DRD5 belonging to the DRD1-like family of the subject to a preset standard level. The DRD1 / 5 level and expression can be increased or decreased bidirectionally by targeting the upstream promoter sequence of DRD1 / 5. The promoter activity of DRD1 / 5 can be attenuated by using antipsychotic drugs that directly target the DRD1 / 5 promoter. The antipsychotic drugs used are typical and / or atypical antipsychotic drugs such as haloperidol, risperidone, clozapine, and cariprazine. This treatment is also useful for subjects in whom the constitutive expression of the DRD1 / 5 gene is impaired. This method can be used to treat schizophrenia, autism, depression, mania, and bipolar disorder by normalizing the DRD1 / 5 expression level. The promoter gene sequencing of DRD1 and / or DRD5 can also be used for quantitative mental health screening by comparing with a standard sequence database.

The present disclosure relates to a novel polynucleotide exhibiting promoter activity, and a method for producing L-isoleucine using same. A microorganism having the novel polynucleotide according to the present disclosure, in which specific positions of promoter regions of an NCgl0302 gene and / or an NCgl0123 gene are mutated, has significantly increased L-isoleucine productivity, and thus the novel polynucleotide can be effectively used to efficiently produce L-isoleucine.

This invention discloses cis-acting elements that enhance the promoter activity of the cotton SWP73B gene and their applications. The invention provides two cis-acting elements that can improve promoter activity, thereby increasing gene transcription levels. These two cis-acting elements are located on the promoter of the cotton boll weight gene GrSWP73B, the sequence of which is SEQ ID NO:1. The cis-acting elements are named GrmotifD and GrmotifE, with their core sequences being CATCA and ATTAA, respectively. The core cis-acting elements GrmotifD and GrmotifE provided by this invention can enhance promoter activity. Vectors containing these cis-acting elements can be used to transform cotton, enabling widespread and efficient expression of the target gene and cultivating new materials with large boll characteristics.

The invention discloses a promoter mutant and an application of the promoter mutant in production of isoleucine. The invention provides a DNA molecule which is any one of the following: A1) a DNA molecule with a nucleotide sequence as shown in SEQ ID No.2; a2) is a variant which is obtained by substitution, deletion or addition of one or more nucleotides at other positions except the 66th nucleotide in the nucleotide sequence as shown in SEQ ID No.2, and the obtained DNA molecule has the same or enhanced promoter activity. Experiments prove that a series of promoter mutants with different strength and regulation characteristics are constructed to replace natural promoters, so that the transcriptional level of a target gene in a key period of fermentation is accurately improved, and the yield of the target gene is increased. The promoter engineering strategy provides a new direction for improving the production performance of industrial microorganisms by optimizing endogenous stress-resistant elements.

The invention relates to the technical field of biological medicines, and particularly discloses application of a swine STING molecular agonist SR-717 in preparation of a pseudorabies virus infection resisting medicine. The swine STING molecular agonist SR-717 can be stably combined with swine STING protein, the promoter activity of a downstream signal molecule interferon regulatory factor 3 (IRF3) and I-type interferon (IFN beta) is enhanced, PRV replication can be remarkably inhibited, the virus load is reduced, and pathological changes of infected mouse lung, brain and other tissues are reduced. The SR-717 provided by the invention has good targeting property and immune activation ability, and is suitable for intervention treatment of porcine pseudorabies virus infection.

The invention provides a method for synthesizing a CMV (cytomegalovirus) promoter and improving the expression quantity of recombinant protein, and belongs to the technical field of gene engineering. The synthetic CMV promoter is based on a CMV promoter, and nucleotide sequences at a transcription factor YY1 binding site are connected in series for 2-7 times; the nucleotide sequence of the CMV promoter is as shown in SEQ ID NO. 1. According to the invention, a transcription factor YY1 and a CMV promoter are subjected to collaborative optimization creatively, the CMV promoter sequence is optimized by analyzing the binding motif of the transcription factor YY1 gene on the CMV promoter sequence, and a plurality of promoter sequences are obtained by connecting binding sites in series. Different experiments prove that the effect of enhancing the activity of the CMV promoter in CHO cells can be achieved, and a new expression element is provided for efficient expression of recombinant protein.

The invention discloses application of a VvMYB APL gene in regulation and control of plant fruit ripening, and belongs to the technical field of plant genetic engineering. The invention finds that the VvMYB APL gene in a grape fruit treated by hydrogen peroxide has a down-regulated expression phenomenon in different stages, which indicates that the VvMYB APL gene may negatively regulate the development of the grape fruit. A VvMYB APL gene is further cloned from 'Kyoho' grapes, subcellular localization shows that the VvMYB APL gene is localized in a cell nucleus, a GUS staining test verifies the promoter activity of MYB APL, and a result shows that the promoter of the gene can start GUS reporter protein, and H2O2 treatment can enhance the activity of the gene. Furthermore, a VvMYB APL gene overexpression and silent vector transformation experiment shows that the silence of the VvMYB APL gene obviously promotes the ripening of grape fruits, and the overexpression inhibits the ripening of the fruits.

The application discloses a molecular genetic marker affecting pig sexual maturity and application thereof. The application takes RBP1 as a research object, and adopts molecular and cell biology methods to study the correlation between RBP1 and the initial estrus period. It is found that six SNP sites in the RBP1 promoter region are related to the initial estrus period. Further research shows that the promoter activity of the CC genotype of the g.80408109T>C site is significantly higher than that of the TT genotype, the promoter activity of the AA genotype of the g.80409632A>G site is significantly higher than that of the GG genotype, and it is confirmed that RBP1 can inhibit the ferroptosis of granulosa cells. It can be seen that the mutation in the RBP1 promoter region can affect the expression of RBP1, and RBP1 can accelerate the initial estrus start by inhibiting the ferroptosis of pig ovarian granulosa cells. The application has good application value for the research on ovarian follicle development and initial estrus start.

The present application discloses an endogenous promoter of Koelreuteria paniculata or Koelreuteria paniculata var. variegata and its application in regulating plant leaf color development. GLK The present application compares the leaf phenotype difference of Koelreuteria paniculata and Koelreuteria paniculata var. variegata, analyzes the chloroplast structure of the two, and determines the photosynthetic characteristic index, finds that the net photosynthetic rate of Koelreuteria paniculata var. variegata is significantly lower than that of Koelreuteria paniculata, and accordingly infers that the gene expression amount of Koelreuteria paniculata var. variegata is down-regulated due to the variation of the gene promoter. GLK Based on the transcriptome data, specific primers are designed to obtain the gene and perform tissue specificity and time sequence development expression pattern analysis, and further, through the FPNI-PCR technology, two types of promoters, A type and B type, are cloned from Koelreuteria paniculata and Koelreuteria paniculata var. variegata, and it is found through analysis that the A type promoter is a promoter common to Koelreuteria paniculata and Koelreuteria paniculata var. variegata, and the B type promoter is a promoter specific to Koelreuteria paniculata; it is found through GUS activity detection that the B type promoter has higher driving activity than the A type promoter. The present application has application prospect in cultivating plant varieties with high photosynthetic efficiency or ornamental plant varieties.

This invention discloses tree fern. AspiMYB5 The application of genes in regulating the biosynthesis of tree fern phenols belongs to the field of genetic engineering technology, which involves regulating the biosynthesis of tree fern phenols within plants. AspiMYB5 The expression level of the gene, thereby regulating the key enzyme gene of tree fern phenol. AspiPKS6 The expression of [a specific enzyme] and the synthesis of tree fern phenol. This invention uses a yeast library screening method to screen genes affecting key enzymes. AspiPKS6 Expressed transcription factors AspiMYB5 The results were verified using yeast single-hybrid technology (Y1H) and electrophoretic mobility variation analysis (EMSA). AspiMYB5 and AspiPKS6 The promoter exhibits direct physical binding in vitro. Dual-luciferase reporter gene assays further demonstrate that... AspiMYB5 and AspiPKS6 When the promoter is co-expressed, the fluorescence signal intensity is significantly reduced, further confirming... AspiMYB5 It can inhibit AspiPKS6 AspiMYB5 Promoter activity. This invention not only improves the theoretical research on the biosynthetic pathway of tree fern phenols, but also provides key target genes and technical support for regulating the content of secondary metabolites in rare plants through genetic improvement.

The application belongs to the field of plant synthetic biology and plant gene expression regulation, and particularly relates to a vector for plant promoter double system quantitative characterization, a standardization method, a product and application. The method combines a protoplast transient expression system and an agrobacterium-mediated leaf transient expression system, uses a compact promoter characterization plasmid (pLIP) containing a firefly luciferase module driven by a to-be-detected promoter and a renilla luciferase internal reference module driven by a 35S promoter, and the two modules are located in a T-DNA interval. The ratio of the two luciferases is calculated, the RPU (relative promoter unit) method is introduced, a standard promoter (35S) is used as a reference, and normalization and direct comparison of promoter activities in different systems are realized. The scheme significantly improves the repeatability and cross-system compatibility of promoter activity data, and is suitable for efficient screening and standardized characterization of regulatory elements in plant synthetic biology.

The invention discloses a regulatory element for enhancing the activity of a promoter and a method for obtaining a high-activity promoter through transformation, and relates to the technical field of gene engineering. The technical problem to be solved by the invention is how to obtain a modified promoter with improved promoter activity. In order to solve the technical problem, the invention provides transcriptional regulation elements from the 1st site to the 1200th site of a nucleotide sequence such as SEQ ID NO: 1. By connecting the transcriptional regulation element to the upstream of a 35S core promoter, a high-activity promoter with the promoter activity 5-6 times that of the 35S promoter can be obtained. According to the research technical scheme, a new insight is provided for analyzing the functional architecture of the corn cis-regulatory element, and a theoretical basis is laid for the rational design of crop synthesis promoters.

Provided is a promoter comprising any one of the following DNAs (1) to (4): (1) DNA comprising a base sequence represented by any one of SEQ ID NO: 1 to SEQ ID NO: 3, (2) DNA comprising a base sequence having a substitution, addition or deletion of one or more bases in the base sequence represented by any one of SEQ ID NO: 1 to SEQ ID NO: 3 and having promoter activity, and (3) DNA comprising a base sequence having a substitution, addition or deletion of one or more bases in the base sequence represented by any one of SEQ ID NO: 1 to SEQ ID NO: 3, (3) DNA having a promoter activity and comprising a base sequence having 90% or more identity with the base sequence represented by any one of SEQ ID NO: 1 to SEQ ID NO: 3, and (4) DNA having a promoter activity and comprising a base sequence capable of hybridizing under stringent conditions with a base sequence complementary to the base sequence represented by any one of SEQ ID NO: 1 to SEQ ID NO: 3.

This invention belongs to the field of plant genetic engineering technology, specifically disclosing a lotus embryo-specific promoter pNnCYP80A and its applications. The nucleotide sequence of the promoter is shown in SEQ ID NO.1. The pNnCYP80A promoter provided by this invention has strong activity and can drive the efficient and specific expression of the GUS reporter gene in the immature embryos of transgenic Arabidopsis thaliana. This promoter can drive the efficient and specific expression of exogenous genes in plant embryo organs, and has broad application prospects in the study of plant embryo organ development and the targeted improvement of embryo quality and yield traits through transgenics.

The application discloses a transcription factor SrMYB1 for regulating synthesis of steviol glycoside, an expression protein thereof and application, and belongs to the technical field of plant genetic engineering. The transcription factor SrMYB1 has a nucleotide sequence as shown in SEQ ID NO. 1, and the amino acid sequence of the expression protein is as shown in SEQ ID NO. 2. Subcellular localization in tobacco epidermal cells shows that the protein coded by the gene is mainly located in the nucleus; yeast activity experiment shows that the protein has transcription activation activity. Yeast single hybridization experiment shows that SrMYB1 can be combined on the promoter of a key gene for synthesis of steviol glycoside SrUGT76G1 . Promoter activity and transgenic experiment both show that SrMYB1 significantly inhibits transcription of the key gene for synthesis of steviol glycoside SrUGT76G1 . Based on the characteristics, certain reference can be provided for quality regulation of steviol glycoside and cultivation of new steviol varieties.

The present invention discloses a single-nucleotide polymorphism (SNP) molecular marker for the FMO3 gene and its application in determining ketosis resistance in cattle, belonging to the technical field of molecular breeding for animal disease resistance. The SNP molecular marker is located at ENSBTAG00000020597: g.-1613T>C. The genotype of this molecular marker is significantly correlated with blood BHB concentration, affecting FMO3 gene promoter activity and, consequently, BHB concentration in the blood of dairy cows. The blood BHB concentration of cows with the CC genotype is significantly lower than that of cows with the TT genotype. Therefore, individual cows with the CC genotype have significantly higher ketosis resistance than those with the TT genotype. This SNP molecular marker can be used in auxiliary breeding of dairy cows to improve the quality of dairy cow populations.

Described herein are nucleic acids having constitutive promoter activity, and the use of such nucleic acids having constitutive promoter activity for expressing a polynucleotide of interest in a plant.

The present application relates to a novel promoter and a method for producing a target product using same, wherein a polynucleotide according to one embodiment has promoter activity, when introduced into a microorganism can increase the expression and activity of an actively linked gene, and can be useful for efficiently producing a target product affected by the gene and the polynucleotide.