43 results about "Cis-regulatory element" patented technology

The invention relates to a high-throughput cis-acting element screening carrier and a screening method. Specifically, the invention provides a plasmid vector system containing bar codes, each bar code in the system is in one-to-one correspondence with a candidate cis-acting element, and the activation multiple of the candidate cis-acting element can be obtained by measuring the abundance of the bar codes; in order to eliminate the influence of a bar code on the vector on a detection result, an exogenous intron which can be cut off during transcription is inserted into a coding region in the vector and is used for distinguishing vector DNA and RNA obtained by transcription. The screening method of the biological cis-acting element has high efficiency, wide applicability and high throughput, and has outstanding application value in biological research and breeding.

To provide non-viral DNA vectors for expressing FVIII therapeutic agents and uses thereof.SOLUTION: The application describes a ceDNA vector having a linear and continuous structure for the delivery and expression of a transgene. The ceDNA vector comprises an expression cassette franked by two ITR sequences, where the expression cassette encodes a transgene encoding FVIII protein. Some ceDNA vectors further comprise a cis-regulatory element including a regulatory switch. Further provided herein are methods and cell lines for reliable gene expression of FVIII protein in vitro, ex vivo and in vivo using the ceDNA vectors.SELECTED DRAWING: None

The invention discloses an application of interaction of a pear transcription factor PbrDDI2 and a PbrAPX14 gene promoter in regulation and control of maturation and senescence of pear fruits. The nucleotide sequence of the pear transcription factor PbrDDI2 is as shown in SEQ ID NO.1 in a sequence table, and the amino acid sequence of the gene coded protein is as shown in SEQ ID NO.2. The invention discloses the transcription factor PbrDDI2 for regulating and controlling the PbrAPX14 gene expression of the pear fruit for the first time. The invention clarifies an action mechanism of interaction of cis-acting elements (TTTGTCG / CGACAAA) in PbrDDI2 and PbrAPX14 gene promoters in clearing endogenous H2O2 accumulation and further inhibiting synthesis of ethylene so as to inhibit fruit ripening and senescence, and provides a theoretical and practical basis for cultivating anti-senescence high-quality pear varieties.

The invention discloses application of a pear transcription factor Pbr3AP2-8 and a gene PbrC3 'H1 in regulation and control of accumulation of chlorogenic acid in pear fruits. The amino acid sequence of the pear AP2 / ERF family transcription factor Pbr3AP2-8 is as shown in SEQ ID NO. 2, and the nucleotide sequence of the encoding gene of the pear AP2 / ERF family transcription factor Pbr3AP2-8 is as shown in SEQ ID NO. 1. The transcription factor Pbr3AP2-8 for regulating and controlling the PbrC3 'H1 gene expression of the pear fruit is disclosed for the first time by analyzing the distribution of cis-acting elements in a PbrC3' H1 promoter and the expression profile of PbrERFs after postharvest storage and ethylene / 1-MCP treatment. The action mechanism of interaction of AP2 binding sites (P: (-466)-(-457)) in Pbr3AP2-8 and PbrC3 'H1 gene promoters in regulation and control of pear fruit chlorogenic acid synthesis is clarified, and a theoretical and practical basis is provided for directional improvement of postharvest pear fruit quality characters.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a promoter of a pinus massoniana terpene synthase gene Pm TPS (-)-alpha-pinene and application of the promoter. According to the invention, a promoter fragment of a Pm TPS (-)-alpha-pinene gene and three homeopathic action elements with jasmonic acid response on the promoter fragment are obtained from pinus massoniana. A complete promoter fragment of the Pm TPS (-)-alpha-pinene gene of the pinus massoniana is transformed into arabidopsis thaliana by means of agrobacterium mediation, and the promoter can drive expression of a GUS gene in leaves of the arabidopsis thaliana or nicotiana benthamiana. GUS staining and GUS enzyme activity of transgenic Bensi tobacco leaves are reduced along with deletion of jasmonic acid response cis-acting elements on the promoter. Furthermore, the yield of pine resin terpenoids can be increased by utilizing the promoter and the homeopathic element thereof.

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.

Provided is a modified virus targeting abnormal ETS family transcription factor-related tumors. The virus comprises a cis-acting element capable of binding to an ETS family transcription factor and a target gene operably linked to the cis-acting element. The virus can be used for treating tumors.

Provided are a nucleic acid construct for improving production of adeno-associated virus and a method for constructing the same, the nucleic acid construct comprising: an adeno-associated virus (AAV) element, a polynucleotide encoding an IE protein; the AAV element comprising a polynucleotide encoding a Cap protein, a polynucleotide encoding a Rep protein, and an AAV cis-acting element. The method for constructing comprises integrating the AAV element carrying an exogenous target gene and the polynucleotide encoding the IE protein into a baculovirus vector skeleton. The recombinant adeno-associated virus (rAAV) obtained by the method has a lower empty shell rate, while improving the rAAV production of a single cell and a unit volume of culture, reducing the production cost, and being easy to scale up.

The invention discloses an APO1 promoter cis-acting element for promoting increase of primary branches of rice panicles and application of the APO1 promoter cis-acting element, and belongs to the technical field of plant genetic engineering. According to the invention, a cis-acting element in the APO1 promoter is excavated, and a CRISPER / Cas9 technology is utilized to prove that the sequence deletion of the cis-acting element can realize moderate increase of primary branches of rice panicles so as to regulate and control panicle types, so that the cis-acting element can be used for rapidly improving the panicle types and the grain number per panicle of rice varieties.

Disclosed herein are gene therapy vectors used for efficiently transducing cells to express a human β-globin gene. Specifically disclosed is an expression vector which comprises: an expression cassette for a β-globin gene, which comprises exons and introns of human β-globin gene, as well as cis-acting elements including one or more of WPRE, SV40 polyadenylation signal and / or SV40 ori. The disclosed expression vectors have significantly enhanced viral vector packaging efficiency in viral vector packaging cell lines, which leads to effective integration of lentiviral vectors and high expression level of β-globin gene in target cells. Also disclosed are pharmaceutical compositions and therapeutic methods utilizing such expression vectors.

The invention relates to a portable organ intelligent computing system. Aiming at an organoid intelligent medical target, a transcription interaction mode jointly regulated and controlled by multiple transcription factors and cis-acting elements is set as input, biological intelligent calculation is performed through the organoid, and output is obtained through a nucleic acid sequencing technology, a pathological detection technology and a space group imaging technology. And analyzing the key gene regulation and control mechanism problem of the organoid intelligent medicine, and assisting the realization of the organoid intelligent medicine.

The application discloses a drought-inducible promoter of Cotinus coggygria and application thereof, a DNA sequence of the promoter is shown as SEQ ID No. 1, proCcDT1 is cloned from a promoter region of a CcDT1 gene of the Cotinus coggygria and is verified through experiments. First, elements of the promoter region are analyzed, and the result shows that the elements contain multiple cis-acting elements that can be involved in response to abiotic stress. Then, proCcDT1-GUS transgenic plants are treated through 10% PEG simulation drought, and it is found that GUS staining of the transgenic plants is enhanced, so it can be concluded that the GUS expression level driven by the promoter is increased under drought induction conditions, and it is also proved that the promoter can be induced by drought stress. The promoter proCcDT1 is introduced into an Arabidopsis genome through genetic transformation technology, so that the promoter can be stably induced by drought stress in a heterologous plant, and the practical application has important value for cultivating drought-resistant Cotinus coggygria varieties.

Provided are a promoter element and the use thereof. The promoter element comprises the nucleotide sequence represented by SEQ ID NO: 17 or one or more sequences having 85% sequence identity thereto. The promoter element can be used as a cis-acting element for expressing a target gene, enhancing the expression of a target protein gene; the promoter element has broad application prospects and high application potential.

A soybean high-temperature inducible promoter and its application. The present invention relates to a soybean inositol galactosidase synthase gene promoter sequence that can be used as a promoter to regulate gene expression under high-temperature stress. The promoter of the present invention is derived from the promoter sequence of the soybean inositol galactosidase synthase gene GmGolS, named GmGolSP, and its base sequence is shown in SEQ ID NO: 1. The sequence contains a variety of stress-related cis-acting elements. The cis-acting elements are MBS, ARE, TC-rich repeats, ABRE, CGTCA-motif and GC-motif. The soybean high-temperature inducible promoter of the present invention is used to induce heat-resistant gene expression under high-temperature stress, and can initiate the efficient expression of heat-resistant related genes under high-temperature conditions, thereby cultivating practical and effective heat-resistant plant varieties. The present invention is applied to the field of plant genetic engineering.

The invention belongs to the field of CRK gene family research, and discloses an identification and expression regulation analysis method of amorphophallus konjac AkCRKs gene family members, which is based on a hidden Markov model of a CRK gene conserved domain, and uses TBtools software to carry out identification and bioinformatics analysis on the AkCRKs gene family members. And the expression characteristics of AkCRKs gene family members in different tissue parts, jasmonic acid, soft rot bacteria (Pcc) and under low temperature stress are analyzed. 19 AkCRKs gene family members are identified in the whole genome of amorphophallus konjac and are non-uniformly distributed on 7 chromosomes. The promoter of the AkCRKs gene family member comprises a plurality of adversity stress response, hormone response, growth and development regulation and other cis-acting elements. The expression of the AkCRKs gene has tissue specificity and participates in response to Pcc, MeJA and low-temperature stress. The research result of the invention provides a basis for further researching the function and adversity stress response mechanism of the amorphophallus konjac AkCRKs gene.

The invention discloses a gene promoter active fragment screening method based on a cross-overlap method and application, and belongs to the technical field of biology. The screening method comprises the following steps: segmenting a target gene promoter sequence, and setting an overlapping fragment between two adjacent segments to obtain a gene promoter active fragment containing the overlapping fragment in the segments. According to the invention, the active fragment of the pennisetum americanum PgGNM2 promoter is screened for the first time, the fragment is used for a CRISPR / Cas9 system, so that the gene editing efficiency can be improved by more than 40%, and when the fragment is used for a yeast one-hybrid bait carrier, the false positive rate can be reduced to less than 5%. The screening method retains the integrity of cis-acting elements, is suitable for monocotyledonous / dicotyledonous plants, and provides an efficient tool for molecular breeding of crops.

The invention discloses identification, characterization and expression analysis of cardamine violifolia WRKY transcription factors and a regulation effect of the cardamine violifolia WRKY transcription factors in selenium accumulation, and belongs to the technical field of biology. WRKY gene family members in cardamine violifolia genes are comprehensively identified, and 120 CvWRKY genes are totally identified. The physical and chemical properties, chromosome localization, phylogeny, gene structure, conserved motif, cis-acting element and colinearity of CvWRKY family gene protein are comprehensively analyzed, and a foundation is laid for understanding the evolutionary relationship and gene function of the CvWRKY gene family. The research finds that the expression level of the CvWRKY34 is in significant positive correlation with the selenium content; meanwhile, both the CvWRKY34 and the CvWRKY021 respond to the induction of Na2SeO3, and the main contribution of the CvWRKY34 and the CvWRKY021 is possibly positive regulation of the selenium absorption process of the root. The research on the cardamine violifolia WRKY family lays a theoretical foundation for analyzing the contribution of the WRKY gene in the high selenium accumulation capacity of the cardamine violifolia, and provides an important clue for functional verification of the cardamine violifolia.

The invention belongs to the technical field of gene engineering, and discloses a recombinant yeast strain for synthesizing astaxanthin as well as a construction method and application of the recombinant yeast strain. The recombinant plasmid provided by the invention comprises the following gene segments: a promoter TEF1p, a beta-carotene hydroxylase gene CrtZ, a cis-acting element IGG6, a beta-carotene ketolase gene CrtW and a terminator CYC1t which are spliced in sequence. A specific cis-acting element IGG6, a promoter, a terminator and exogenous CrtZ and CrtW are selected, a gene segment capable of producing astaxanthin is obtained, a recombinant plasmid is constructed from the gene segment, the recombinant plasmid is transformed into a saccharomyces cerevisiae strain, and high transformation of astaxanthin is achieved. Furthermore, the CrtZ and the CrtW are mutated by adopting error-prone PCR (Polymerase Chain Reaction) to obtain forward mutation, so that the conversion efficiency of the astaxanthin is further improved, and a more economic and more efficient way is provided for heterologous production of the astaxanthin by a microbial chassis.

The application provides a MhPHR2-MhSCL29 double-gene co-transformation synergistic regulation module and application, and belongs to the field of microbial genetic engineering.In the application, the phosphate-starvation response factor MhPHR2 is used as an upstream regulation hub, directly binds to a P1BS cis-acting element on a promoter of a downstream GRAS family transcription factor MhSCL29, positively activates the transcription of the MhSCL29, and forms a complete phosphate signal-mycorrhizal symbiosis regulation cascade.The nucleotide sequence of MhPHR2 in the synergistic regulation module is as shown in SEQ ID NO.1, and the nucleotide sequence of MhSCL29 is as shown in SEQ ID NO.2.The application improves the mycorrhizal symbiosis potential and the phosphorus utilization efficiency of a stock from the root.

The invention discloses a cotinus coggygria drought inducible promoter and application thereof, the DNA sequence of the promoter is as shown in SEQ ID No.1, proCcDT1 is cloned from a cotinus coggygria CcDT1 gene promoter region, and experimental verification is carried out. Firstly, promoter region elements are analyzed, and a result shows that the promoter region elements contain a plurality of cis-acting elements possibly participating in response to abiotic stress. Afterwards, a proCcDT1-GUS transgenic plant is subjected to 10% PEG (polyethylene glycol) simulated drought treatment, and the GUS dyeing degree of the transgenic plant is enhanced, so that the GUS expression level driven by the promoter under a drought induction condition is increased, and the promoter is proved to be induced by drought stress. The promoter proCcDT1 is introduced into an arabidopsis thaliana genome through a genetic transformation technology, so that the promoter proCcDT1 can be stably induced by drought stress in heterologous plants, and the practical application of the promoter proCcDT1 has important value for cultivating drought-tolerant cotinus coggygria varieties.

The present invention discloses the application of bovine SP110 gene splicing editing in enhancing the anti-tuberculosis ability of cattle, belonging to the technical field of bovine anti-tuberculosis breeding. The present invention starts from splicing-related cis-acting elements, and regulates the alternative splicing reaction of bovine SP110 by introducing cis-acting elements, thereby affecting the anti-tuberculosis ability of cattle. The cis-acting element sequence is named the pre-SAND-exon sequence, which is located in exon 12 and the adjacent 5'-terminal intron 12 of the human or equine SP110 gene, and the sequences are shown in SEQ ID NO.1 and SEQ ID NO.2. The present invention first regulates the alternative splicing of the bovine SP110 gene by introducing the pre-SAND-exon sequence into the bovine SP110 gene, reduces the expression abundance of the spliceosome bSP110c with lower anti-tuberculosis ability, and regulates the higher abundances of the spliceosomes bSP110a and bSP110b with anti-tuberculosis infection ability. The provided bovine SP110 gene splicing editing can effectively improve the anti-tuberculosis ability of cattle, can be used for the breeding of anti-tuberculosis cattle, effectively simplifies the breeding method, improves the breeding efficiency, and saves the breeding cost.

The invention discloses a method for screening, identifying and functionally analyzing a poplar latex protein family at a whole genome level by using multiple bioinformatics methods, and relates to the field of plant gene identification and screening. The method comprises the following steps: performing preliminary screening, re-modeling, re-screening and identification on an obtained poplar whole genome file, an annotation information file, a protein sequence file and an MLP protein family structure file to obtain a final poplar MLP gene family member; on-line physicochemical property analysis software is used for carrying out predictive analysis on physicochemical properties, and predictive software is used for carrying out possible function analysis on a gene structure, a conservative structural domain, a promoter region cis-acting element and protein subcellular localization; evolutionary trees of the poplar MLP family members and the arabidopsis MLP family members are constructed through evolutionary tree software, and evolution analysis and classification of the poplar MLP family members and the arabidopsis MLP family members are analyzed. According to the method, the interested poplar MLP family gene can be effectively and quickly screened, and the time and cost of molecular means screening are greatly reduced.

The invention discloses a cotton verticillium wilt-resistant gene identification method based on family analysis and application thereof, and 89 LRX family members are identified in four cotton genus by carrying out whole genome identification and analysis on a cotton LRX gene family; through evolutionary analysis, the LRX gene family is divided into four subgroups, the LRX genes in the same subgroup show similar conserved motifs, but some genes show significant differences; through chromosome position analysis and collinearity analysis, the LRX gene family experiences many fragment repetition events and cis-acting element analysis shows that the gene family is likely to participate in various biological and abiotic stress responses. Through an expression mode under the stress of verticillium wilt, a virus-induced gene silencing experiment is carried out by utilizing the GhLRX7, and a gene silencing plant shows more serious disease symptoms, so that the GhLRX7 plays an important role in the aspect of verticillium wilt resistance of cotton.

The invention relates to the technical field of plant genetic engineering, and provides a ramie high-temperature inducible promoter and application thereof, and the nucleotide sequence of the promoter is as shown in SEQ ID NO: 1. The ramie high-temperature inducible promoter provided by the invention is rich in cis-acting elements (CCAAT-Box) related to temperature, also has elements related to drought (LTRE), dehydration (CATNTG motif) and wound induction (AG motif), and can be used as an effective tool for creating high-temperature-resistant plant varieties through genetic engineering.

The invention belongs to but is not limited to the technical field of gene analysis, and particularly relates to an activity analysis method of a tea tree F3 'H promoter, which comprises the following steps: S1, cloning F3' HP1 and a 5'end fragment deletion promoter sequence thereof, and constructing an expression vector; s2, analyzing the cis-acting element of the F3 'HP1 and the 5'end fragment of the F3' HP1 without the promoter sequence; s3, carrying out GUS histochemical staining and GUS enzyme activity determination on the instantly converted tobacco; and S4, response of the F3 'H to low-temperature treatment. The result shows that the promoter sequence (F3' HP1) of the upstream 1806bp of the F3 'H gene' ATG 'is obtained through cloning, and four 5' fragment-deleted F3 'H gene promoter sequences, namely F3' HP2, F3 'HP3, F3' HP4 and F3 'HP5 are obtained, and the sequence lengths are 804, 479bp, 195 and 154bp respectively, the F3' HP1 sequence also has response elements related to low temperature, light, gibberellin and the like besides basic elements of TATA-box and CAAT-box, and the transcriptome data result shows that the F3 'H responds to low temperature.

This invention discloses the application of the HbCBF1 gene in regulating ethylene content in rubber trees, belonging to the field of agricultural biotechnology. By overexpressing the HbCBF1 gene (nucleotide sequence shown in SEQ ID NO.1), this invention significantly enhances the biosynthesis of ethylene in transgenic plants, manifested as an increased ethylene release rate, elevated content of the ethylene precursor ACC, and enhanced activity of key enzymes in the ethylene synthesis pathway, SAMS, ACS, and ACO. It also exhibits dwarfing phenotypes associated with excessive ethylene accumulation, including significantly reduced plant height, thinner stems, shorter internodes, and accelerated leaf senescence. Molecular mechanism studies show that HbCBF1 directly activates the transcriptional expression of SAMS1 by binding to the CRT / DRE cis-acting element in the SAMS1 gene promoter, thereby upregulating ethylene biosynthesis. This invention provides a new target gene and regulatory method for the genetic engineering improvement of rubber trees.

The invention discloses an application of cattle SP110 gene splicing editing in enhancing the anti-tuberculosis ability of cattle, and belongs to the technical field of cattle anti-tuberculosis breeding, starting from splicing related cis-acting elements, the variable splicing reaction of the cattle SP110 is regulated by introducing the cis-acting elements, so that the anti-tuberculosis ability of the cattle is influenced. The sequence of the cis-acting element is named as a pre-SAND-exon sequence, the cis-acting element is located on the 12th exon of a human or horse SP110 gene and the 12th intron adjacent to the 5'end of the human or horse SP110 gene, and the sequences of the cis-acting element are shown as SEQ ID NO.1 and SEQ ID NO.2. According to the invention, the pre-SAND-exon sequence is introduced into the cattle SP110 gene for the first time, so that the variable splicing of the cattle SP110 gene is regulated and controlled, the expression abundance of a splicing body bSP110c with relatively low anti-tuberculosis capability is reduced, and the abundance of bSP110a and bSP110b splicing bodies with anti-tuberculosis infection capability is regulated and controlled to be higher. The provided cattle SP110 gene splicing editing can effectively improve the anti-tuberculosis ability of cattle, can be used for breeding anti-tuberculosis cattle, effectively simplifies the breeding method, improves the breeding efficiency, and saves the breeding cost.

The present invention discloses the promoter of Neolamarckia cadamba sucrose transporter gene NcSUT1 and its application. The promoter sequence of 2015 bp upstream of the NcSUT1 gene was cloned from Neolamarckia cadamba. Cis-acting element analysis found that it contains multiple core promoter elements such as TATA-box and CAAT-box, as well as stress response, phytohormone and light response elements. According to the positions of the promoter functional elements, expression vectors of the full-length promoter and 6 5'-terminal deletion fragments were constructed. The results showed that the promoter fragment from -1377 bp to -1 bp upstream of NcSUT1 could drive the expression of LUC in tobacco leaves, and the fragment from -669 bp to -1 bp had the strongest promoter activity. Transgenic Arabidopsis analysis showed that the NcSUT1 promoter could drive the expression of GUS in the parts rich in vascular tissues in tissues such as leaf veins and roots, petals, anthers, filaments, flower stalks, siliques, funicles, and fruit pod stalks of seedlings, and the expression was specific. The present invention not only provides candidate sequences for screening promoters specifically expressed in vascular parts, but also can accurately achieve the germplasm innovation and genetic improvement of fast-growing forest trees.

The invention discloses an application of a cis-acting element of rice or corn in regulating and controlling the heading stage of the cis-acting element, the cis-acting element is located in a florigen gene regulation and control region of the rice or the corn, and the nucleotide sequence of the cis-acting element is' ATCTGAT 'or' ATCAGAT '. By mutating or deleting the cis-acting element sequence, the heading period of rice or corn can be properly delayed, the heading period of gramineous plants such as rice and corn can be regulated and controlled, and rice and corn varieties with delayed heading periods can be obtained. The invention has important significance for improving the adaptability of gramineous plant varieties such as rice and corn to different regions and seasons.