74 results about "Promoter activity" patented technology

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 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 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 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 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 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.

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.

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. A polynucleotide according to one embodiment has promoter activity, can be introduced into a microorganism to enhance the expression and activity of a gene operably linked thereto, and can be effectively used for efficiently producing a target product that is affected by the polynucleotide and the gene.

The application discloses a pig beta-defensin-3 gene core promoter and a construction method and application thereof, and belongs to the technical field of genetic engineering. The sequence of the core promoter is shown as SEQ ID No. 1, the core promoter has significant promoter activity in pig small intestinal epithelial cells, and can be regulated by nutrients such as sodium butyrate and glutamine. The application also discloses a construction method for specifically amplifying the core promoter, which comprises the following steps: performing PCR amplification by taking pig small intestinal epithelial cell DNA as a template, taking the upstream primer and the downstream primer shown as SEQ No. 2-SEQ No. 3 as the primer, constructing a pMD-18T-pBD-3-P recombinant plasmid, and taking the recombinant plasmid as a template to amplify a core promoter fragment. The core promoter provides an excellent experimental system for studying the transcriptional regulation mechanism of the pig beta-defensin-3 gene, the nutrient response characteristics provide a new platform for studying the interaction between nutrition and immunity, and the core promoter has a wide application prospect in the field of healthy breeding of livestock and poultry.

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.

The invention provides a promoter pchsA derived from petunia, a gene expression cassette of betacyanin and application of the gene expression cassette, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the promoter pchsA is shown as SEQ ID No.1; according to the invention, a petunia organ specific promoter pchsA is utilized to construct a gene expression cassette of the betacyanin, so that heterologous synthesis of the betacyanin is successfully driven in tobacco, and expression and accumulation of the betacyanin in tissues of roots, stems, leaves and flowers of the tobacco are realized. According to the invention, a new transgenic tobacco germplasm capable of efficiently accumulating the betacyanin is created, a new material is provided for plant bioreactor production of the high-value pigment (betacyanin), and meanwhile, a method for rapidly, intuitively and visually detecting the activity of the promoter is established.

The invention discloses a pancreatic tissue specific expression recombinant adeno-associated virus vector and application thereof, and belongs to the technical field of biological medicine. Comprising the following steps: S1, cloning a pancreatic specific promoter: jointly using Ins2 and Pdx1 promoters to prepare a Pdx-1 / Ins2 promoter; performing serotype and capsid mutation optimization; s2, insertion of an exogenous gene expression cassette: cloning a target gene to the downstream of a Pdx-1 / Ins2 promoter to construct a'promoter-exogenous gene-polyadenylic acid signal 'expression unit; s3, regulation and control element integration: inserting an HS4 insulator sequence into two sides of an expression box, and adopting Gibson assembly to realize seamless connection; constructing an SIN structure: deleting a U5 region in ITR through site-specific mutagenesis or enzyme digestion, and verifying that no residual promoter activity exists; s4, virus packaging and purification: packaging rAAV particles by adopting an HEK293 cell three-plasmid co-transfection system; the three plasmids comprise a carrier plasmid, an auxiliary plasmid and a packaging plasmid; and determining the purity after purification.

The invention discloses a bovine SLC27A6 gene promoter vector as well as a construction method and application thereof, and belongs to the field of molecular biology and genetic engineering. According to the invention, the bovine SLC27A6 gene promoter SLC27A6-A-SLC27A6-E of which the nucleotide sequences are respectively shown as SEQ ID NO.1-SEQ ID NO.5 is obtained through cloning, and bovine SLC27A6 gene promoter vectors are respectively constructed by taking a pGL4.10 vector as a skeleton vector. Experimental results show that in MAC-T cells, the bovine SLC27A6 gene promoter vector constructed by the invention can exert promoter activity and regulate and control downstream gene expression. The invention provides a new biological material for regulation of gene expression in bovine cells and genetic breeding of the targeted SLC27A6 gene, and has a wide application prospect.

The present invention relates to a nucleic acid having promoter activity in cone 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 said expression cassette, vector, viral particle, cell or pharmaceutical composition in the treatment of eye diseases, in particular hereditary retinal diseases.

This invention discloses VvENO2 The application of genes in regulating plant fruit development belongs to the field of plant genetic engineering technology. Previous research in this invention found that hydrogen peroxide (H2O2) treatment significantly promotes grape fruit ripening. Transcriptome analysis of H2O2-treated grapes revealed… VvENO2 Gene expression levels decreased. To further clarify... VvENO2 The role of the gene in plant fruit development was investigated through subcellular localization experiments and promoter activity analysis, demonstrating its nucleus location and that its promoter can activate the GUS reporter protein. Furthermore, treatment with ABA and H2O2 enhanced its activity. In addition, transgenic Arabidopsis and transgenic grape were constructed, and phenotypic observations confirmed... VvENO2 Genes negatively regulate the ripening of plant fruits. This invention provides a new regulatory site for breeding early-maturing plant varieties.

The invention discloses a core promoter for construction of a gene expression system in mammalian cells and application of the core promoter, and provides polynucleotide with promoter activity, a transcription expression cassette containing the polynucleotide, a recombinant expression vector, a recombinant host cell and application in an expression system. The core promoter provided by the invention can be combined with various enhancers and coding sequences in a modular manner, is suitable for constructing a drug-induced expression system, a tumor specific expression system, a synthesis loop and a gene expression regulation system related to cell therapy, and has good universality and application prospects.

The invention discloses a construction method of a mouse model for monitoring AMR activity and / or AMR promoter activity in the liver in real time, and belongs to the field of animal models and construction methods thereof. The invention aims to solve the technical problem of how to realize in-vivo, non-invasive and / or dynamic monitoring of AMR activity and / or AMR promoter activity in the liver. Therefore, the invention provides a method for constructing an animal model, the method comprises the following steps: a DNA molecule is introduced into a receptor animal, and the DNA molecule comprises a reporter gene and an AMR promoter for promoting transcriptional expression of the reporter gene; the receptor animal contains an AMR gene and / or the AMR promoter. The small animal model can be used for evaluating the regulating effects of different factors on AMR activity and / or AMR promoter activity in vivo, can also be used as an evaluation model of the regulating effects of cells, drugs and the like on liver AMR activity and / or AMR promoter activity, and can be widely applied to various biomedical experiments.

The present application relates to a novel promoter and a method for producing target materials using the same. More specifically, the present application relates to a novel polynucleotide having promoter activity, a gene expression cassette, and a host cell comprising the same, and a method for producing target materials using the microorganism.

The invention provides a beef cattle MSTN gene promoter function optimization design method based on deep learning and model adaptation. The beef cattle MSTN gene promoter function optimization design method comprises the following steps: taking RNA-seq expression quantity of an MSTN gene as a supervision label, and taking high-dimensional epigenetic signal prediction data obtained by inputting a promoter region sequence into a pre-trained Enformer deep learning model as an input feature; training a machine learning adapter model; when the prediction precision of the adapter model reaches a preset threshold value, determining the adapter model as a verification model; constructing a promoter variant sequence library of the MSTN gene by adopting a multi-strategy variant generation method; the promoter activity is verified by a reporter gene expression verification method aiming at the promoter sequence which ranks in the top in the variant sequence library. The invention also discloses an enhanced promoter with a sequence as shown in SEQ ID NO.4 and a weakened promoter with a sequence as shown in SEQ ID NO.2 or 3.

The invention discloses a large yellow croaker interferon i gene promoter sequence and application thereof. By analyzing genome data of the pseudosciaena crocea, the pseudosciaena crocea IFNi gene promoter is cloned, and the sequence of the pseudosciaena crocea IFNi gene promoter is provided, and the sequence is as shown in SEQ ID NO.1. Experiments prove that the sequence has promoter activity, and the activity of the sequence is significantly up-regulated under the induction of a viral nucleic acid simulant Poly (I: C). In addition, the interferon regulatory factor 1 (IRF1) can also be used for remarkably inducing the activity of the IFNi gene promoter of the large yellow croaker. Cloning and activity identification of the promoter provide a good experimental system for researching expression regulation and control of the IFNi gene of the large yellow croaker, create conditions for constructing an efficient expression vector to express an exogenous gene or applying the expression vector to transgenic fish construction by using the promoter, and have important theoretical and practical significance.

This invention discloses the promoter EP11 and related biomaterials and applications, belonging to the field of biotechnology. The technical problem solved by this invention is how to increase the content of L-lysine. The promoter EP11 disclosed in this invention has the sequence shown in SEQ ID No. 1 of the sequence listing. The EP11 promoter is introduced into *Corynebacterium glutamicum*, a bacterium capable of synthesizing L-lysine, thereby driving the expression of genes in the L-lysine synthesis pathway, resulting in recombinant *Corynebacterium glutamicum*. Culturing this recombinant *Corynebacterium glutamicum* yields L-lysine, and the production of L-lysine is increased. This demonstrates that the EP11 promoter of this invention not only possesses promoter activity but can also be used for the production of L-lysine.