5207 results about "Promoter" patented technology

The present application relates to a kind of deletion mutant nucleic acid and its application in anti-herbicide.The deletion mutation occurs in the promoter of rice OsHPPD Gene, the length of deletion sequence is at least 10 bp, and the A in the start codon ATG of gene is 0, at least the bases located in the upstream of start codon ATG from 2041 to 2032 are deleted. OsHPPD The present application finds that by deleting part of the sequence in the promoter of rice OsHPPD Gene, rice can obtain resistance to HPPD inhibitor herbicides, which is of great application value for ensuring agricultural production safety and improving the efficiency of herbicide use.

The invention discloses a streptomycete recombinant engineering strain for producing mycosporine-like metabolites, a gene expression cassette is integrated in a genome of the streptomycete recombinant engineering strain, the gene expression cassette is recombined to an attB site of a chromosome in a streptomycete chassis strain, the gene expression cassette is an operon controlled by a single promoter, and the operon is an operon controlled by a single promoter. The operon comprises 2-4 biosynthetic gene elements in MysA, MysB, MysC and MysD or mysDp334 in a synthetic pathway of mycosporine-like metabolites. The invention also discloses a construction method of the streptomyces recombinant engineering strain and an application of the streptomyces recombinant engineering strain in preparation of the mycosporine-like metabolites. A mycosporine-like metabolite is efficiently produced by heterologous expression of a mycosporine-like metabolite synthesis gene element in streptomyces, and four mycosporine-like metabolite products with high yield and strong ultraviolet absorption activity can be obtained by reconstruction of the biosynthesis gene element and recombinant expression in streptomyces pluvialis.

The invention relates to the field of nucleic acid detection, in particular to a multifunctional dumbbell type nucleic acid probe, a kit, a target detection method and application thereof. The multifunctional dumbbell type nucleic acid probe provided by the invention comprises two or more of the following functional areas: a promoter area used for being combined with RNA polymerase; the target recognition area is used for recognizing and combining a target sequence and opening the dumbbell type nucleic acid probe; the dumbbell type nucleic acid probe can be transcribed and combined with a rolling ring, a specific sequence is subjected to mass transcription, amplification of the specific sequence is achieved, finally, signal amplification is achieved during detection, and the dumbbell type nucleic acid probe has the advantages of being simple in reaction system, low in background signal and high in detection specificity; and the dumbbell type nucleic acid probe can be designed according to different target sequences, so that the controllable detection of different targets can be realized.

The invention discloses a mammalian cell expression vector, an expression plasmid and an expression system. The expression vector sequentially comprises the following elements: (a) a promoter; (b) a Kozak sequence; (c) a polyadenine sequence, the polyadenine sequence sequentially comprises a bGH PolyA sequence and a 5 'UTR PolyA sequence, the nucleotide sequence of the bGH PolyA sequence is as shown in SEQ ID NO.5, and the nucleotide sequence of the 5' UTR PolyA sequence is as shown in SEQ ID NO.6; (d) copying the start site; and (e) screening marker genes. According to the present invention, the expression vector is optimized by the expression element, such that the exogenous gene can be conveniently, efficiently and flexibly inserted and edited by the expression vector so as to achieve the stable extracellular or intracellular expression of the exogenous protein, and the expression efficiency of the exogenous protein can be improved by carrying out the expression plasmid construction based on the expression vector; and more efficient expression and purification of the foreign protein are realized.

The invention relates to a deleted mutant nucleic acid and application thereof in herbicide resistance. The deletion mutation occurs in a promoter of a rice OsHPPD gene, the length of a sequence of a deleted nucleic acid is at least 25 bp, A in an initiation codon ATG of the OsHPPD gene is taken as a 0 site, and at least bases at 2299 to 2275 sites on the upstream of the initiation codon ATG are deleted. According to the application disclosed by the invention, the rice can obtain resistance to HPPD inhibitor herbicides by deleting part of sequences in the promoter of the rice OsHPPD gene, so that the application has great application values in guaranteeing agricultural production safety and improving the use efficiency of the herbicides.

The invention discloses an endogenous tRNA (transfer ribonucleic acid) promoter. The nucleotide sequence of the endogenous tRNA promoter is shown as SEQ ID NO: 1; an expression cassette containing the endogenous tRNA promoter and a recombinant vector; the invention also relates to application of the endogenous tRNA promoter, the expression cassette or the recombinant vector in thraustochytriaceae microorganism gene editing, and a thraustochytriaceae microorganism gene editing method. The CRISPR / Cas9 gene editing system based on the endogenous tRNA promoter is successfully established, convenient, rapid and efficient fixed-point editing in thraustochytriaceae microorganisms is achieved, and the CRISPR / Cas9 gene editing system has high application value; a transformation system for screening the recombinant thraustochytriaceae microorganisms by using sodium acetate as an auxotroph is established, the transformation screening efficiency of the recombinant thraustochytriaceae microorganisms obtained based on an agrobacterium transformation method is remarkably improved, and rapid gene editing is assisted.

The invention relates to a deleted mutant nucleic acid and application thereof in herbicide resistance. The deletion mutation occurs in a promoter of the rice OsHPPD gene, the length of a deletion sequence is at least 10 bp, A in an initiation codon ATG of the OsHPPD gene is taken as a 0 site, and at least 2041-2032 sites of basic groups located on the upstream of the initiation codon ATG are deleted. According to the application disclosed by the invention, the rice can obtain resistance to HPPD inhibitor herbicides by deleting part of sequences in the promoter of the rice OsHPPD gene, so that the application has great application values in guaranteeing agricultural production safety and improving the use efficiency of the herbicides.

The invention belongs to the technical field of crop molecular markers, and particularly discloses a molecular marker related to the waterlogging tolerance of brassica napus and application of the molecular marker. According to the invention, a 1.2 Kb transposon insertion and a 2.6 Kb transposon insertion are identified by utilizing GWAS (Good White Attenual Sequence) and are positioned in a promoter region of the Bna.C08. GUN4 gene, and the rape containing the two transposon insertion has high waterlogging resistance. The molecular marker is used for auxiliary selection of oilseed rape with strong waterlogging resistance, whether the oilseed rape germplasm is waterlogging-resistant oilseed rape germplasm can be predicted only by detecting the characteristic band of the molecular marker, the identification method is simple and feasible, the selection efficiency is high, and the molecular marker has huge application potential in the field of oilseed rape waterlogging resistance improvement breeding.

The invention provides a plant salt tolerance related transcription factor TaVOZ1 as well as a coding gene and application thereof, and belongs to the technical field of functional genes. The TaVOZ1 is related to plant salt tolerance and has an amino acid sequence as shown in SEQ ID No. 1, and a gene for coding the protein has a nucleotide sequence as shown in SEQ ID No. 2. The plant salt tolerance can be regulated by regulating the content or activity of the protein or the expression quantity of the gene, for example, when the gene is over-expressed, the plant salt tolerance is enhanced, and when the gene is knocked down, the salt tolerance is weakened. The TaVOZ1 disclosed by the invention can be combined with 5 '-CTTCT-3'or a reverse complementary sequence 5'-AAGAAG-3 'in a downstream target gene promoter to activate the expression of a stress response gene so as to improve the salt tolerance of a plant, so that the TaVOZ1 can be used for culturing a salt-tolerant wheat variety.

The invention discloses a long-chain non-coding RNA molecular marker related to bovine semen quality and application of the long-chain non-coding RNA molecular marker, long-chain non-coding RNA comprises an LNC68393 promoter sequence and an LNC68393 gene sequence, a molecular marker SNP1 is located on the promoter sequence, and a molecular marker SNP2-5 is located on the gene sequence. Different polymorphisms of the molecular marker are significantly related to the bull semen quality, and the molecular marker can be applied to detection of the bull semen quality, further early evaluation of the bull reproductive performance, cultivation of excellent bulls, shortening of the breeding time, improvement of the efficiency and reduction of the cost, and has important application value in improvement of the reproductive performance of the whole bull herd.

The invention relates to the field of gene therapy, in particular to an RBM25 gene epigenetics activation technology based on a CRISPR / dCas9-SunTag system and application of the RBM25 gene epigenetics activation technology in ischemic heart failure treatment, the RBM25 gene epigenetics activation technology comprises the following steps: (a) protein containing inactivated Cas9 (dCas9) fused with a plurality of GCN4 peptide repetitive sequences, the number of the GCN4 peptide repetitive sequences is 8-12, and the dCas9 contains D10A and H840A double mutations; (b) a protein containing fusion of an anti-GCN4 single-chain antibody fragment (scFv) and a TET1 catalytic structural domain; (c) at least one gRNA of a target RBM25 gene promoter region, wherein a target sequence of the gRNA is selected from a sequence as shown in SEQ ID NO: 1-3; and (d) a lipid nanoparticle (LNP) system for delivery of said (a), (b), and (c).

The preparation method comprises the following steps: firstly, analyzing the amino acid sequence of an alpha 1 chain of the human XVII type collagen, and selecting partial amino acid fragments which are not easily degraded by endogenous or exogenous protease in non-transmembrane regions of different functional domains of the protein; the recombinant collagen gene fragment (Col17a) n is obtained by repeatedly splicing the recombinant collagen gene fragment (Col17a) n; then introducing homologous promoters from different methylotrophic yeasts to the 5'end of a recombinant collagen gene segment, optimally synthesizing selected (Col17a) n and promoter segments, obtaining a connecting segment by utilizing PCR cloning, connecting the collagen segment (Col17a) n, the promoter segments and a pPIC9K carrier skeleton segment by virtue of a Gibson Assessment seamless connection technology, and obtaining the recombinant collagen gene. And constructing to obtain the pichia pastoris expression plasmid pPIC9K-(Col17a) n of the recombinant human source XVII type collagen.

The invention provides a grass carp sourced U6 promoter. The sequence of the grass carp sourced U6 promoter is shown as one of SEQ ID NO: 1-3. Three grass carp-derived U6 promoters are cloned, the transcriptional activity and cross-species adaptability of the three promoters are analyzed, results show that the three promoters can show transcriptional activity in fish cells and human cells, the transcriptional activity of ciU6-1 is remarkably higher than that of the other two promoters, heterologous DNA can be transcribed in various cells, and the expression of the ciU6-1 in the fish cells and the human cells is improved. And the method can be used for gene editing means based on Cas9.

The invention discloses a coconut strong promoter, a plant expression vector and application. The nucleotide sequence of the promoter is as shown in SEQ ID NO. 1. Compared with a known promoter, the promoter provided by the invention can drive high-efficiency expression of related genes, and is a strong promoter. The invention also relates to a plant expression vector containing the strong promoter and application of the plant expression vector in promoting expression of a target gene.

The present disclosure relates to compositions and methods for reducing expression of MYC gene in a cell. In some embodiments, an expression repressor comprises a targeting moiety that binds a MYC promoter, anchor sequence, or super-enhancer. In some embodiments, the expression repressor comprises an effector moiety that represses transcription or methylates DNA. Systems comprising two expression repressors are also disclosed. The compositions can be used, for example, to treat cancers such as HCC.

The disclosure provides polynucleotides containing regions of the Myosin 15 (Myo15) promoter, as well as vectors containing the same, that can be used to promote expression of a transgene specifically in hair cells. The polynucleotides described herein may be operably linked to a transgene, such as a transgene encoding a therapeutic protein, so as to promote hair cell-specific expression of the transgene. The polynucleotides described herein may be operably linked to a therapeutic transgene and used for the treatment of subjects having or at risk of developing hearing loss or vestibular dysfunction.

The invention discloses a darkness-induced enhanced luminescence expression vector as well as a construction method and application thereof, and the enhanced luminescence expression vector is constructed by taking a vector pDES as a skeleton vector. A core element formed by sequentially connecting a promoter PsDIN6, a mutated nnLuz gene mnnLuz, a HisspS gene of neon lamp bacteria, an H3H gene, a CPH gene and an NPGA gene of aspergillus nidulans is inserted between an attR1 element and an attR2 element of a vector pDES from the 5'end to the 3 'end, and the vector is obtained. A luciferase Luz gene of luminous fungus neon lamp bacteria is subjected to random mutation by using a directed evolution technology, a luciferase mutant gene mLuz of which the luminous efficiency is 5.7 times higher than the original luminous efficiency is obtained through screening, and an expression vector constructed by the luciferase mutant gene mLuz is instantaneously expressed on tobacco leaves, so that the tobacco leaves do not emit light in daytime and emit bright fluorescence in darkness.

Described herein are synthetic promoters and / or enhancers that are specific for cancer cells and methods of engineering synthetic cancer-specific promoters.

The invention belongs to the technical field of bioengineering, and provides a recombinant XVII type collagen as well as a preparation method and application thereof. A target gene sequence is obtained through bioinformatics predictive analysis and integrated design on the basis of a human XVII type collagen sequence, codon optimization is carried out, a T7 promoter in a plasmid vector is optimized, an engineering strain of the high-hydrophobicity humanized XVII type collagen is prepared, and the high-hydrophobicity humanized XVII type collagen is obtained. The strain is subjected to culture and inducible expression to obtain high-yield recombinant XVII type collagen. The recombinant XVII type collagen disclosed by the invention is reasonable in sequence design, large in expression quantity, simple in preparation method and stable in heredity, and rapid separation of the collagen can be realized in a production and preparation process by utilizing high hydrophobicity.

The invention belongs to the field of gene engineering and biological treatment, and provides a hypoxia specific gene expression cassette, gram-negative bacteria containing the expression cassette and application of the gram-negative bacteria in preparation of tumor drugs. According to the invention, the forward hypoxia promoter and the reverse hyperxia promoter are reasonably designed, and appropriate survival essential genes are matched, so that the hypoxia specific gene expression cassette which is more efficient and safer is obtained. Gram-negative bacteria modified by the hypoxia specific gene expression cassette designed by the invention are low in overall mutation rate, and in a mouse in-vivo tumor model, more bacteria are distributed in tumors and the clearing speed in normal organs is higher.

The invention discloses application of a transcription factor CgGATA8 in adjusting lignin synthesis, and belongs to the technical field of genetic engineering breeding and fruit and vegetable preservation. According to the invention, the transcription factor CgGATA8 is instantaneously transferred into the juice sac of the Majia pomelo through an agrobacterium infection method, the lignin content in the over-expressed juice sac is obviously higher than that in no-load control, and autofluorescence signals are enhanced, secondary cell walls are thickened and the juice sac granulation symptom is aggravated in phloroglucinol dyeing and paraffin section at the same time. Molecular biology function verification shows that the Marijia pomelo transcription factor CgGATA8 can be combined with and activate a CgPOD16 promoter, the expression level of the CgPOD16 gene is improved, and the accumulation amount of lignin synthesis in fruits is improved, so that the CgGATA8 gene participates in regulation and control of synthesis and deposition of lignin in Marijia pomelo fruit juice cells, and has the function of promoting maturity and aging of picked fruits.

The invention provides a shikimic acid pathway artificial operon and a construction method and application thereof.According to the artificial operon, shikimic acid pathway related genes are integrated and connected in different modes, a trc promoter and an RBS ribose binding site sequence are optimized, and the artificial operon formed through combination has the advantages of being stable, efficient and wide in applicability; different from a relatively extensive shikimic acid way transformation mode in the past, the operon can obviously improve the accumulation capacity of aromatic amino acid, and when the operon is applied to an L-phenylalanine production strain, the yield of L-phenylalanine reaches 105 g / L, and the saccharic acid conversion rate reaches 26.5%.

The invention provides a lentivirus for treating cancer as well as a preparation method, application and a corresponding pharmaceutical composition of the lentivirus. According to the invention, an hTERT promoter and a CDC6shRNA gene sequence are inserted behind an enzyme cutting site of a vector, an obtained plasmid is transferred into an expression strain, the expression strain is cultured and screened to obtain a cloned strain, and the cloned strain is shaken and packaged to obtain the hTERT-CDC6shRNA lentivirus; when the lentivirus disclosed by the invention is used for infecting cancer cells, the experimental result shows that the proliferation of the cancer cells can be effectively inhibited.

The invention belongs to the field of gene engineering, and particularly relates to a nucleic acid molecule for changing corn plant height and application thereof. A series of nucleic acid molecules capable of changing the corn plant height are obtained by editing a corn ZmGA2ox2 gene promoter.

The invention discloses corynebacterium glutamicum for producing N-acetylglucosamine and application of the corynebacterium glutamicum, and belongs to the technical field of synthetic biology. Specifically, the corynebacterium glutamicum overexpresses a GNA1 gene from a brewer's yeast S288C controlled by a tacM promoter, a glmS gene from corynebacterium glutamicum S9114 and a yqab gene from escherichia coli K12 MG1655, and simultaneously has five gene defect types of nagA, nagB, ldhA, msccg and msccg2 in a genome and two gene single base mutation types of odhA and glnA, and the codon of the glnA for coding the 405th amino acid is mutated from TAC to TTC. By blocking excretion of glutamic acid of corynebacterium glutamicum and enhancing conversion of glutamic acid to glutamine, the supply capacity of a precursor of corynebacterium glutamicum is enhanced, and an optimal way for synthesizing N-acetylglucosamine is obtained, so that efficient production of N-acetylglucosamine in corynebacterium glutamicum is realized, and the yield of N-acetylglucosamine in corynebacterium glutamicum is increased. The method has important economic value and social significance.

The invention discloses a soybean bidirectional promoter and application thereof, and belongs to the technical field of separation and application of bidirectional promoters. The nucleotide sequence of the soybean bidirectional promoter disclosed by the invention is as shown in SEQ ID NO. 2. Experiments prove that the soybean bidirectional promoter can bidirectionally and simultaneously drive the expression of a target gene, and by applying the soybean bidirectional promoter, plant traits can be improved, transgenic plants or new plant varieties can be cultivated, and the biological breeding process can be promoted.

The invention relates to a gene editing vector of agrobacterium, a gene editing method of the gene editing vector and an agrobacterium editing strain. The agrobacterium gene editing vector comprises the following gene elements: a plasmid skeleton, a Pvan promoter, a spCas9 gene, a green fluorescent protein gene GFP, gRNA, and an upstream homologous arm and a downstream homologous arm of a target gene. According to the agrobacterium gene editing vector, a CRISPR / spCas9-GFP gene editing system is adopted, an efficient, simple, convenient and traceless genome editing system is constructed in agrobacterium, and gene knockout and insertion can be achieved.

The invention relates to a chimeric promoter and application thereof. Nucleotide sequences of the chimeric promoter comprise: (1) a nucleotide sequence having at least 95% sequence identity with a nucleotide sequence shown in SEQ ID NO: 1; and (2) a nucleotide sequence as shown in SEQ ID NO: 1. The chimeric promoter provided by the invention has high activity, and can significantly increase the expression level of a target heterologous nucleotide sequence in plants.

The invention discloses a preparation method of a recombinant engineering bacterium for expressing XVII type collagen COL17A1, and relates to the technical field of gene engineering, in particular to a preparation method of a recombinant engineering bacterium for expressing XVII type collagen COL17A1, which comprises the following steps: gene synthesis and vector construction: synthesizing an optimized sequence SEQ ID No.1 of human source XVII type collagen COL17A1; adding proper restriction enzyme cutting sites at two ends of the sequence; the synthesized gene sequence is cloned to a pGAPZ A-alpha A vector, and the expression of the gene under the control of a GAP promoter is ensured. According to the preparation method of the recombinant engineering bacterium for expressing the XVII type collagen COL17A1, by using a GAP promoter, the constructed recombinant engineering bacterium can keep a stable expression level under different environmental conditions, the GAP promoter serving as a constitutive promoter can continuously and stably express recombinant protein in pichia pastoris, an inducer does not need to be added, the production process is simplified, and the production cost is reduced. The production cost is reduced, and the production efficiency is improved.

The invention relates to the technical field of plant genetic engineering, in particular to a promoter and application thereof in orchid plants. The invention provides an orchid endogenous promoter which is a powerful constitutive promoter and can drive a target gene to be powerfully expressed in various tissues such as roots, stems, leaves and flowers of an orchid. Compared with a CaMV 35S promoter, the promoter has the advantages that the driving efficiency of the gene in orchid plants is greatly improved, an important tool is provided for molecular breeding of plants, and the promoter has a relatively good application prospect.