27 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.

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 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.

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.

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 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 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.

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 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.

Disclosed herein are gene therapy vectors for efficient transduction of cells to express human beta globin genes. Specifically disclosed is an expression vector comprising: an expression cassette of a beta globin gene comprising an exon and an intron of a human beta globin gene, and a cis-acting element comprising one or more of WPRE, an 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 results in efficient integration of lentiviral vectors in target cells and high expression levels of beta globin genes. Pharmaceutical compositions and methods of treatment utilizing such expression vectors are also disclosed.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a grape VvWRKY70 gene and application of the grape VvWRKY70 gene in regulation and control of plant drought resistance. In order to excavate a grape drought-resistant gene and research the regulation and control mechanism of the grape drought-resistant gene, the invention screens out a VvWRKY70 gene which is subjected to drought-induced expression, the nucleotide sequence of the VvWRKY70 gene is shown as SEQ ID NO: 1, and the VvWRKY70 gene encoding protein is positioned in a cell nucleus, can specifically recognize a W-box cis-acting element and shows transcriptional activation activity. The VvWRKY70 gene positively regulates the drought resistance of plants, and overexpression of the gene can improve the relative water content and survival rate of plant leaves, promote accumulation of the content of an osmotic regulation substance proline and the content of a secondary metabolism substance anthocyanin, reduce the content of malondialdehyde and the content of hydrogen peroxide and remarkably enhance the tolerance of the plants under drought stress.

The invention relates to the field of plant biology, and discloses application of a transcription factor GhMYB42 in cotton plant type improvement, the nucleotide sequence of the transcription factor GhMYB42 is shown as SEQ ID No.1, and the cotton plant type is improved by introducing the transcription factor GhMYB42 into cotton cells and combining with a cis-acting element in a cotton target gene promoter. A transcription factor GhMYB42 is introduced into cotton cells, so that after the GhMYB42 is expressed in the cotton cells, the GhMYB42 is specifically combined with a cis-acting element in a cotton target gene promoter to regulate and control the expression of a target gene, and then key plant type characters such as the plant height, the internode length, the leaf area, the fruit branch included angle and the fruit branch length of cotton are directionally regulated and controlled. The cotton plant type is converted into an ideal mechanical cotton picking plant type which is short in plant height, short in internode, small in leaf area, small in fruit branch included angle and short in fruit branch.

The application discloses an adenine base editor capable of realizing precise deletion of a small DNA fragment and construction and application thereof. The adenine single-base editor comprises a fusion protein ABE8e-EndoV, and the fusion protein comprises, from N terminus to C terminus, an adenine deaminase, a nickase variant of a Cas nuclease and an isosine I base deletion repair endonuclease in sequence. The adenine base editor for precise deletion of a small DNA fragment has the characteristics of not only high efficient A-G base substitution, but also 9-13 bp small fragment deletion from the deaminated A base to the cleavage site interval of the Cas nuclease protein. The adenine base editor is particularly suitable for mining and research of cis-acting elements in genomes of animals and plants, and has wide application prospects in single-base substitution of target genes and saturation mutation screening.

The invention provides a method for identifying drought-resistant key AGO genes in haloxylon ammodendron and haloxylon ammodendron, and relates to the technical field of gene identification. According to the method for identifying drought-resistant key AGO genes in haloxylon ammodendron and haloxylon ammodendron, 45 HaAGO genes in haloxylon ammodendron and 30 HpAGO genes in haloxylon ammodendron are respectively identified through hmmscan screening, homologous comparison and CDD / PFAM structural domain verification; then, through protein physicochemical property analysis, phylogenetic tree construction, conservative motif and gene structure analysis, chromosome localization, promoter cis-acting element prediction and protein interaction network analysis, molecular characteristics and evolution laws of an AGO gene family are determined, and finally, in combination with transcriptome sequencing data under a wet / drought habitat, the AGO gene family is determined. And screening out the gene which is obviously up-regulated under drought stress. The invention provides a systematic method for mining drought-resistant gene resources of haloxylon ammodendron and haloxylon ammodendron, and the screened key gene can be used as a target for drought-resistant molecular breeding and has important significance on ecological restoration of northwest desert regions.

The invention relates to a modified virus targeting ETS family transcription factor abnormality related tumors, which comprises a cis-acting element capable of being combined with ETS family transcription factors and a target gene operably connected with the cis-acting element, and can be used for treating tumors.

The present application relates to the technical field of plant genetic engineering, and provides a ramie high-temperature inducible promoter and application thereof, wherein the nucleotide sequence of the promoter is shown as SEQ ID NO:1. The ramie high-temperature inducible promoter is rich in temperature-related cis-acting elements (CCAAT-Box), and further 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.

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.

The invention relates to the technical field of biology, in particular to a wheat TaTT8 gene related to wheat anthocyanin biosynthesis and application of a promoter of the wheat TaTT8 gene. The invention provides application of a protein TaTT8 or a biological material related to the protein TaTT8, and the application is any one of Y1-Y3: Y1, application in regulating and controlling the color of wheat grains; y2 in increasing the anthocyanin content of wheat; y3 is applied to cultivation of purple wheat or cultivation of wheat with high anthocyanin content. A TaTT8 overexpression wheat material is obtained to prove that TaTT8 is a key factor of color change of purple wheat seed coats, but a TaTT8 gene coding sequence in purple wheat is not different from that in white wheat, and a specific cis-acting element exists in a TaTT8 gene promoter region in the purple wheat to improve expression of TaTT8, so that biosynthesis of wheat anthocyanin is promoted, and the yield of wheat anthocyanin is increased. Purple wheat germplasm resources are innovated, and the method is of great significance to improvement of grain quality.

The invention provides a recombinant pinellia ternata lectin expression vector, a construction method and application thereof, and belongs to the technical field of biology. According to the invention, RNA (Ribonucleic Acid) of pinellia ternata is successfully extracted, synthesis of a first chain of cDNA (Complementary Deoxyribonucleic Acid) is completed, and nine pinellia ternata lectin genes PTA1-1, PTA1-2, PTA1-3, PTA1-4, PTA2, PTA3, PTA4, PTA5 and PTA6 are successfully cloned. Nine pinellia ternate agglutinin gene promoter fragments are successfully cloned, and a cis-acting element analysis result shows that the pinellia ternate agglutinin gene promoter contains core promoter elements CAAT-box and TATA-box as well as cis-acting elements such as light response, hormone response, stress correlation, plant growth and development regulation and the like, so that a new thought is provided for attenuation of pinellia ternate; and a theoretical basis is provided for medicine development based on the pinellia ternate agglutinin. According to the construction method disclosed by the invention, six pinellia ternate agglutinin expression vectors, namely pET-28a (+)-PTA1-1, pET-28a (+)-PTA1-2, pET-28a (+)-PTA1-3, pET-28a (+)-PTA1-4, pET-28a (+)-PTA5 and pET-28a (+)-PTA6, are successfully constructed.