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126 results about "Transcriptional expression" patented technology

CRISPR/Cpf1 plant genome directional modification function unit, carrier comprising function unit, and application thereof

The invention belongs to the technical field of gene engineering, and concretely relates to a CRISPR / Cpf1 plant genome directional modification function unit, a carrier comprising the function unit, and application thereof. The invention aims to build an effective plant genome CRISPR / Cpf1 directional modification framework carrier according to an action principle of a CRISPR / Cpf1 system and a vegetable cell gene expression characteristic, and realize high-efficient application thereof in plant genome directional modification. The technical scheme provided by the invention is characterized by building the CRISPR / Cpf1 plant genome directional modification framework carrier formed by lachnospiraceae bacterium Cpf1 nuclease protein (LbCpf1) expression unit and a guiding crRNA transcription expression unit. The invention also provides a method for designing specificity-guided crRNA and building a CRISPR / Cpf1 recombinant expression vector aiming at rice genome target loci by adopting the CRISPR / Cpf1 plant genome directional modification framework carrier. The invention provides the high-efficient CRISPR / Cpf1 plant directional modification framework carrier, which can effectively realize simple, fast and high-efficient directional modification based on the CRISPR / Cpf1 system aiming at a plant genome target sequence.
Owner:UNIV OF ELECTRONIC SCI & TECH OF CHINA

Method for rapidly detecting grouper irido virus

The invention discloses a fast detection method for grouper iridovirus, firstly proposing a loop-mediated isothermal nucleic acid amplifying detection applied for the detection of grouper iridovirus, which comprises the steps as follows: (1)a ORF-014L is cloned and a recombinant plasmid template containing ORF-014L is constructed; (2)primer design, primer sequence; (3)amplification, the reaction temperature is 63-65 DEG C, and the reaction time 20-60mins. The LAMP detection technique of the invention can rapidly detect SGIV, the transcriptional expression of SGV 014L in cells, the grouper tissue infected by SGIV, and the GP cells infected by SGIV; the sensitivity of the invention is three orders of magnitude higher than Nested PCR, reaching to six point six copies.
Owner:SUN YAT SEN UNIV

Plant stress-tolerance-associated protein TaDREB3A and coding gene and application thereof

The invention discloses a plant stress-tolerance-associated protein TaDREB3A and a coding gene and application thereof. The protein provided by the invention is one of the following proteins: (a) a protein consisting of an amino acid sequence shown as a sequence 1 in a sequence list, and (b) a protein which is associated with plant stress tolerance and is derived from the sequence 1 by substituting and/or deleting and/or adding one or several amino acid residues for the amino acid sequence shown in the sequence 1. The TaDREB3A provided by the invention can express under the induction of drought, high salt, high temperature, low temperature, pathogenic bacteria, ABA (Abscisic Acid), ethylene, JA (Jasmonic Acid) and SA (Salicylic Acid), and can specifically regulate and control transcription expression of the gene containing a DRE/CRT (Dehydration-Responsive Element/C-Repeat) cis-element (core sequence: CCGAC); therefore, the drought resistance, the salt tolerance, the high temperature resistance and the resistance to the powdery mildew pathogen of the plant are improved. The TaDREB3A provided by the invention provides a base for artificially controlling the expression of genes related to the stress tolerance and the stress resistance, and is about to play an important role in cultivation of plants with enhanced stress tolerance and enhanced stress resistance.
Owner:INST OF CROP SCI CHINESE ACAD OF AGRI SCI

Plant stress tolerance associated protein GmNF-YA17, and encoding gene and application thereof

Plant stress tolerance associated protein GmNF-YA17, and an encoding gene and an application thereof. The protein is a protein (a) composed of an amino acid sequence represented by sequence 1 in a sequence table, or a protein (b) obtained by substituting and / or deleting and / or adding one or more amino acid residues to the protein (a), associating with the plant stress tolerance and derived from the sequence 1. Experiments prove that gene GmNF-YA17 can be obtained through cloning from soybeans, is expressed under induction of drought, salt, ABA and ethylene, positions the coded protein to a nucleus, and can specifically regulate the transcription expression of gene containing a CCAAT-box cis-element. The GmNF-YA17 can improve the drought resistance of plants, provides a foundation for artificial control of expression of stress resistance and stress tolerance associated genes, and plays a great role in the cultivation of stress resistance and stress tolerance reinforced plant.
Owner:INST OF CROP SCI CHINESE ACAD OF AGRI SCI

Mesothelin-targeted replication-defective recombinant lentivirus CAR-T transgenic carrier as well as establishment method and application thereof

The invention discloses a Mesothelin-targeted replication-defective recombinant lentivirus CAR-T transgenic carrier which comprises a pronucleus replicon pUC Ori sequence for plasmid replication, an amicillin resistance gene AmpR containing sequence for mass amplification of target strain, virus replicon SV40Ori sequence for enhancing replication in eukaryocyte, a lentivirus packaging cis-element for lentivirus packaging, a ZsGreen1 green fluorescent protein for green fluorescence expression of eukaryocyte, an IRES ribosome combination sequence for joint transcriptional expression of protein, a human EF1(alpha) promoter for the eukaryotic transcription of chimeric antigen receptor gene, a chimeric antigen receptor for forming a second-generation CAR or third-generation CAR integrating identification, transfer and start, and an eWPRE element for improving the transgenic expression efficiency. Moreover, the invention also discloses an establishment method and application of the carrier. In the invention, the secretion of cell factors and the in-vitro killing effect of CAR-T cells can be remarkably enhanced, and the effect of clinical treatment of malignant pleural mesothelioma and pancreatic cancer is outstanding.
Owner:SHANGHAI UNICAR THERAPY BIOPHARM TECH CO LTD

Plant disease-resistant protein GhWRKY40, coding gene and application thereof

The invention discloses a protein, the name of which is GhWRKY40, which is a protein shown in (a) or (b) or (c): (a) a protein with the amino acid sequence shown in sequence 2 in a sequence table; (b) a fusion protein obtained by connecting tags at the N end or / and the C end of the protein shown in the sequence 2 in the sequence table; and a protein with the function or activity of GhWRKY40 transcription factors which is obtained by deriving the protein shown in (a) or (b) through substitution, deletion or / and insertion of one or more amino acids. The gene has the advantages that the GhWRKY40 regulates transcription expression of specific genes, causes physiological and biochemical changes of plants, increases resistance of plants to pathogen invasion, so as to protect growth and development of plants, and has important significance in application of improving response of plants to pathogens.
Owner:九圣禾种业股份有限公司

Method for regulating and controlling growth of chrysanthemum petals through conversion of CmTCP20 gene

The invention belongs to the field of plant genetic engineering and transgenic breeding and relates to a method for regulating and controlling growth of chrysanthemum petals through conversion of the CmTCP20 gene.The method includes the following steps that the CmTCP20 gene is cloned from florist's chrysanthemum 'ShenMa'; a plant expression vector of the CmTCP20 gene is constructed; the plant expression vector is transferred into chrysanthemum through an agrobacterium-mediated method, culturing is conducted, and a resistant plant is preliminarily obtained; the converted plant is subjected to PCR and quantitative RT-PCR detection to verify that the CmTCP20 endogenous gene is integrated to a genome DNA of the transgenic plant and subjected to transcription; the transgenic plant is subjected to phenotypic observation and statistic analysis, and it is found that compared with a plant not subjected to transgenosis, the petal growth amount of the transgenic plant is obviously increased.According to the method, the endogenous CmTCP20 transcription factors are converted and subjected to normal transcriptional expression, so that growth of the chrysanthemum petals is regulated and controlled, a novel and practical method is provided for improving ornamental quality of chrysanthemum through a genetic engineering technology, and the chrysanthemum biotechnology breeding process can be effectively promoted.
Owner:NANJING AGRICULTURAL UNIVERSITY

Human iPS cell gene editing and screening method

The invention discloses a human iPS cell gene editing and screening method. The method comprises the following steps: introducing editing plasmids into iPS cells by an electroporation method; producing gene-edited cells through resistance screening and enrichment; and performing low-density passage, and selecting monoclonal cell colonies for enlarged culture and identification. An EF1 alpha promoter is used for promoting free plasmids capable of simultaneously promoting Cas9 and screening resistance protein expression in human iPS cells, after the plasmids enter the human iPS cells, Cas9 protein and resistance protein can be transiently and stably expressed in the cells at the same time, and in the most vigorous time period of plasmid transcriptional expression, screened substances are added into a culture environment. The iPS cells which obtain exogenous plasmids and express the resistance protein survive, besides, the expressed Cas9 protein is guided by sgRNA to be subjected to double-strand cleavage at the target sequence position of a genome for gene editing, and the cells which do not obtain the exogenous plasmids or are not expressed by the plasmids die due to the fact that the cells do not contain resistance. Use of a flow cytometry or insertion of exogenous genes into the cell genome is avoided.
Owner:UNION STEMCELL & GENE ENG

Thinopyrum intermedium ERF-transcription factor and its coding gene and use

The invention discloses a middle couch grass ERF transcription factor and its coding gene. The middle couch grass ERF transcription factor can be the protein with one of the following amino acid residues sequences that SEQ ID No: 2 in sequence list; or its amino acid residues sequence which is replaced and / or missed and / or added with one or many amino acid residues, and relevant to plant against disease against reverse performance. The transcription expression of the coding gene is greatly induced by rhizoctonia cerealis, salt, cold, ethylene etc. The TiERF1 of the invention supplies the base for the relevant gene expression, gene resources for gene engineering breeding, and will play an importance role in increasing plant against disease against reverse performance for gene engineering breeding.
Owner:INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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