87 results about "Cis acting" patented technology

Disclosed are nucleic acid vectors which comprise: (a) a transfer nucleotide sequence comprising (i) a plant active promoter, operably linked to (ii) a recombinant tobacco rattle virus (TRV) cDNA (preferably derived from TRV RNA2) which includes at least cis acting elements permitting replication of the cDNA; a subgenomic promoter operably linked to a sequence encoding a TRV coat protein; and a heterologous nucleotide sequence which is foreign to the virus;(b) border sequences which permit the transfer of the transfer nucleotide sequence into a plant genome. Such vectors may be used as expression vectors or for achieving viral induced gene silencing (VIGS) of a target gene, wherein the heterologous nucleotide sequence is a targeting sequence which corresponding to that gene. Example vectors include pTV00 and vectors which are derived from PTV00 and have the characteristics thereof. Also disclosed are associated processes, methods, viruses or viral particle, kits, host cells and plant tissues.

An extensible RNA-based framework for engineering ligand-controlled gene regulatory systems, called ribozyme switches, that exhibit tunable regulation, design modularity, and target specificity is provided. These switch platforms typically contain a sensor domain, comprised of an aptamer sequence, and an actuator domain, comprised of a hammerhead ribozyme sequence. A variety of modes of standardized information transmission between these domains can be employed, and this application demonstrates a mechanism that allows for the reliable and modular assembly of functioning synthetic hammerhead ribozyme switches and regulation of ribozyme activity in response to various effectors. In some embodiments aptamer-regulated cis-acting hammerhead ribozymes are provided.

A novel nucleic acid construct for down-regulating angiogenesis in a tissue of a subject is provided. The nucleic acid construct includes: (a) a first polynucleotide region encoding a chimeric polypeptide including a ligand binding domain fused to an effector domain of an apoptosis signaling molecule; and (b) a second polynucleotide region encoding a cis acting regulatory element being for directing expression of the chimeric polypeptide in a specific tissue or cell; wherein the ligand binding domain is selected such that it is capable of binding a ligand present in the specific tissue or cell, whereas binding of the ligand to the ligand binding domain activates the effector domain of the apoptosis signaling molecule. Also provided are methods of utilizing this nucleic acid construct for treating diseases characterized by excessive or aberrant neo-vascularization or cell growth.

The present application describes isolated nucleic acids that contain a Nic gene product responsive element, and the use thereof in methods of producing transgenic tobacco plants having reduced levels of nicotine and / or TSNA therein, as well as other plants or host cells that contain altered levels of a protein of interest therein due to inclusion of a cis-acting decoy element therein.

A helper dependent adenoviral vector system is provided. The subject helper dependent adenoviral vector system is made up of: (1) a "gutless" adenoviral vector that include cis-acting human stuffer DNA that provides for in vivo long term, high level expression of a coding sequence present on the vector; (2) an adenoviral helper vector that is characterized by having an adenoviral genome region flanked by recombinase recognition sites, where the helper vectors further include a non-mammalian endonuclease recognition site positioned outside of the adenoviral genome region; and (3) a mammalian cell that expresses the corresponding recombinase and endonuclease, as well as the adenoviral preterminal and polymerase proteins. Also provided are methods of using the subject systems to produce virions having the subject helper dependent adenoviral vectors encapsulated in an adenoviral capsid. In addition, kits for use in practicing the subject methods are provided.

The invention relates to a new gene for the rice zinc-finger protein transcription factor and the application thereof to drought resistance and salt tolerance, in particular to the rice zinc-finger protein transcription factor, a coding sequence thereof, a carrier or host, a cis-acting element and antagonist of the rice zinc-finger protein transcription factor or the coding sequence. The rice zinc-finger protein transcription factor comprises polypeptide, and the polypeptide can be the polypeptide having the amino acid sequence of SEQ ID NO: 2, the conservative variant polypeptide or the homologous polypeptide separated from the same; the carrier or host includes the coding sequence; and the cis-acting element is combined with the rice zinc-finger protein transcription factor. The invention also relates to a method for improving the drought resistance and the salt tolerance of the rice and a method for sieving the rice with high drought resistance and salt tolerance. The novel method for improving and studying the drought resistance and the salt tolerance of the rice has wide application prospect.

A novel nucleic acid construct for down-regulating angiogenesis in a tissue of a subject is provided. The nucleic acid construct includes: (a) a first polynucleotide region encoding a chimeric polypeptide including a ligand binding domain fused to an effector domain of an apoptosis signaling molecule; and (b) a second polynucleotide region encoding a cis acting regulatory element being for directing expression of the chimeric polypeptide in a specific tissue or cell; wherein the ligand binding domain is selected such that it is capable of binding a ligand present in the specific tissue or cell, whereas binding of the ligand to the ligand binding domain activates the effector domain of the apoptosis signaling molecule. Also provided are methods of utilizing this nucleic acid construct for treating diseases characterized by excessive or aberrant neo-vascularization or cell growth.

The present invention provides compositions and methods for regulating expression of heterologous nucleotide sequences in a plant. Compositions include a novel promoter nucleotide sequence for the gene encoding ubiquitin in Tripsacum dactyloides, as well as vectors, microorganisms, plants and plant cells comprising the promoter nucleotide sequence, or variants and fragments thereof. Methods for expressing a heterologous nucleotide sequence in a plant using the promoter sequences disclosed herein are also provided. The methods comprise stably incorporating into the genome of a plant cell a nucleotide sequence operably linked to the promoter of the present invention and regenerating a stably transformed plant that expresses the nucleotide sequence.

A method for using the promotor LEAP of rice's drought inducting gene to improve the resistance of plant to drought features that the promotor LEAP of rice OsLEAl gene is cloned. Its nucleotide sequence is shown by SEQ ID No.1 and is used to code a drought induced expression protein. Two cis-acting elements ABRE contained in its promotor region can be specifically response to drought and ABA. Its inductive expression carrier can stongly induce the expression of target gene when the drought stress is applied to a plant. The method for culturing the drought-resistant plant is also disclosed.

The invention discloses an induced slow virus expression system which comprises a target gene expression cassette and an rtTA expression cassette. The target gene expression cassette comprises an induced promoter containing a tetracycline cis-acting response element and a target gene, and the rtTA expression cassette comprises a promoter, an rtTA, a linker peptide and a screening gene, wherein a P2A linker peptide is adopted as the linker peptide, the screening gene is Puro, and the induced promoter is TetO6 or TRE3G. The invention further discloses a construction method of the induced slow virus expression system. The construction method comprises the steps of pLVX-Puro carrier reconstructing; TetON3G gene synthesizing and cloning; pLVX-rtTA3 constructing; EF1a promoter subcloning; red fluorescence protein (RFP) subcloning; subcloning of the induced promoter TRE3G or TETO6 containing the tetracycline cis-acting response element. The induced slow virus expression system can carry out inducible expression of the target gene to the greatest level while keeping low background expression after cells are introduced into the system, is sensitive in response for an inducer, high in efficiency and low in molecular weight and can be widely applied to gene function research.

A helper dependent adenoviral vector system is provided. The subject helper dependent adenoviral vector system is made up of: (1) a "gutless" adenoviral vector, which in certain embodiments includes cis-acting human stuffer DNA that provides for in vivo long term, high level expression of a coding sequence present on the vector, where in certain embodiments the vector includes an integrating domain; (2) an adenoviral helper vector that is characterized by having an adenoviral genome region flanked by recombinase recognition sites, where the helper vectors further include a non-mammalian endonuclease recognition site positioned outside of the adenoviral genome region and in certain embodiments a third adenoviral inverted terminal repeat (ITR) sequence positioned between first and second terminal ITRs; and (3) a mammalian cell that expresses the corresponding recombinase and endonuclease, as well as the adenoviral preterminal and polymerase proteins. Also provided are methods of using the subject systems to produce virions having the subject helper dependent adenoviral vectors encapsulated in an adenoviral capsid. In addition, kits for use in practicing the subject methods are provided.

The invention provides a novel HBB overexpression vector and a design method and application thereof. The HBB overexpression vector comprises an HBB expression module; the HBB expression module comprises a DNase I core high-sensitivity site, a promoter, an HBB expression frame and a downstream high-sensitivity site which are arranged in series; the DNase I high-sensitivity site comprises HS4, HS3,HS2, 3'E and the like which are expressed in series. The total length of the HBB expression module is smaller than 4 kb. By simplifying and optimizing a cis-acting element and the HBB expression frame, not only is the length of the HBB expression module obviously shortened, but also the transcriptive activation strength of the HBB expression module is improved, and efficient, stable and specificactivation of the HBB globin gene is realized.

The invention relates to an induction-enhanced tissue specific promoter and application thereof, belonging to the technical field of plant genetic engineering. A promoter (BPHP, i.e. Bph14 promoter) for Oryza Sativa L. genes resistant to brown plant hoppers is separated from Oryza Sativa L. and is cloned. The promoter has a nucleotide sequence shown as SEQ ID No. 1, can control Oryza Sativa L. genes resistant to the brown plant hoppers to be specifically expressed in vascular bundle tissues and can be expressed in a way that the feeding induction of the brown plant hoppers is enhanced. The promoter contains BS1EGCCR cis-acting elements and can enable the genes to be specifically expressed in plant vascular bundle tissues. Moreover, the promoter has defense-related response cis-acting elements which can be combined with WRKY transcription factors, and the gene expression is increased when Oryza Sativa L. is fed by the brown plant hoppers or is infected by pathogenic bacteria. The promoter can be used as the promoter for the specific expression in the vascular bundle tissues and can be used as the brown plant hopper and pathogenic bacteria induction-enhanced promoter.

The invention pertains to the technical field of plant genetic engineering, and particularly relates to a promoter CBFP of a key gene CbCBF of a shepherd spurse CBF path and applications thereof. The nucleotide sequence of the promoter is showed as SEQ ID NO:1, wherein CbCBF gene and the promoter thereof contain the nucleotide sequence of the 2283rd basic group in the sequence showed in the sequence table of SEQ ID NO:1, and the nucleotide sequence encodes a transcription factor which is induced by low temperature and can regulate and control a cold-inductive gene. The promoter area contains two ABRE cis-acting elements which are serially arranged, thus being capable of specifically playing a response reaction to low temperature and ABA; and the promoter area contains three MYCRE cis-acting elements CANNTG, thus being capable of being combined with an upstream control element (ICE) (inducer of CBF expression). An inductive expression carrier structured by using the genetic promoter can powerfully induces the expression of a target gene when the plant is stressed by low temperature. The invention also provides the applications of the promoter in cultivating cold-resistant plants, which realizes the breed improvement of crops.

A novel mini-retrotransposon vector system is provided for integrating foreign DNA into plants. The invention includes a novel vector comprising a truncated and modified retroelement which includes the 5′ and 3′ LTR regions that provide transcription initiation and termination sites as well as the cis acting sequences required for reverse transcription. Novel vectors, plant cells, and methods of using the same are disclosed.

The invention discloses an ERF transcription factor related to the plant stress resistance and an encoding gene and application thereof. The invention firstly provides an ERF transcription factor gene LcERF056 separated from lotus japonicas, the amino acid sequence of the gene LcERF056 is shown as SEQ ID No.1, and the nucleotide sequence of the encoding transcription factor is shown as SEQ ID No.2. The invention further provides an ERF transcription factor structure domain which can be combined with a cis-acting element to start stress resistance response gene expression and is shown as SEQ ID No.3. Function analysis experiments prove that the resistance of a plant to high salt stress can be effectively enhanced or improved by overexpressing the LcERF056 gene in the plant, and therefore the protein encoded by the LcERF056 gene has the functions of the EFR transcription factor. The invention further provides application of the ERF transcription factor and the encoding gene on the aspects of enhancing the adversity stress resistance of the plant, breeding new transgenic plant varieties with the adversity stress resistance and the like.

The invention relates to the field of medicaments, in particular to a novel bifunctional cicatrix and tissue fibrosis resistant oligonucleotide medicament. The nucleic acid contains an AP-1 cis-acting component in high affinity binding with AP-1 and a Smad cis-acting component in high affinity binding with Smad.

The invention discloses a promoter. The promoter can regulate and control the specific expression of a gene in secretion-type glandular hair, and a nucleotide sequence of the AaALDH1 gene promoter is shown as SEQ ID NO.1. The invention also provides an application of the promoter in the genetic engineering breeding by adopting plant secretion-type glandular hair tissues to express and produce metabolite. In addition, the invention also discloses a carrier and an expression cassette containing the promoter. By adopting the AaALDH1 gene promoter, a foundation is set for researching the expression regulation and control of the AaALDH1 gene and analyzing a cis acting element on the promoter, and the important significance on the genetic engineering breeding utilizing the plant secretion-type glandular hair tissues to express and produce the metabolite also can be realized.

A recombinant plasmid or expression vector comprising a sequence encoding a trans-acting hairpin ribozyme or inserted RNA flanked by 5′ and 3′ self-cleavage cis-acting hairpin ribozymes, which produces a long RNA transcript that undergoes self-catalyzed cleavage at the 5′ and 3′ sides of the trans-acting ribozyme or inserted RNA.

The invention discloses a main cis-acting element of a shrimp white spot syndrome virus (WSSV) iel promoter and a transcription factor combined with the same and application. In the invention, by starting from transcriptional regulation of WSSV iel and carrying out structural and functional analysis on the promoter of the WSSV iel through deletion and mutation, a 12-bp DNA is found to be the maincis-acting element of the WSSV iel and is a crucial factor for the high expression of the iel. In the invention, a DNA affinity chromatography method is used for purifying a protein combined with a DNA segment from a nucleus protein Sf9, the protein is identified to be PHB2 (Poly-Beta-Hydroxybutyrate 2) through biological mass spectrometry, and the interaction between the protein and the DNA segment is proved to be specific by an electrophoretic mobility shift assay. Experiment results prove that PHB2 serves as a transcription factor and is specifically combined with a 12-bp DNA sequence of the iel promoter to start WSSV immediate early gene transcription so as to further regulate the replication of the WSSV, and can be used as an effective action target of medicaments for screening medicaments for resisting the shrimp WSSV.

A gene capable of increasing an expression level of aspergillus niger xylanase, a recombinant plasmid and a host cell thereof relate to optimal codon optimization, GC content adjusting and optimization of cis-acting element and sequential-repetitive sequence on a xylanase gene from aspergillus niger. According to design of a site-directed mutagenesis primer, artificial reconstruction on an original aspergillus niger xylanase gene is carried out to obtain an xylanase gene xynBT over-expressed in pichia pastoris. The reconstructed aspergillus niger xylanase gene xynBT has an expression level in pichia pastoris increased by 2.6 times, compared with that of an original gene xynB; during high density fermentation in a 3 L fermentation cylinder, the xylanase gene xynBT has an expression level in pichia pastoris reaching 20424.2 U / mL. The invention can be applied to molecule reconstruction on the xylanase gene and production of a recombinant xylanase, and can substantially increase the expression level of the xylanase.

The present invention relates to an isolated nucleic acid sequence comprising a first DNA sequence comprising a cis-acting replication element (CARE) from an Adeno-Associated Virus (AAV), and a second DNA sequence operably linked to said CARE, wherein amplification of said isolated nucleic acid sequence occurs when said isolated nucleic acid sequence is integrated in the genome of a cell and said cell is contacted with a CARE-dependent replication unducer (CARE-DRI). It also relates to amplification methods using a CARE-dependent replication inducer (CARE-DRI) and packaging cell-lines wherein replication of the integrated rep and cap genes is inducible by a CARE-DRI.

The invention discloses a system for rapidly analyzing an RNA (ribonucleic acid) functional element in vivo. The system comprises a bifluorescence reporter vector p35mG, and the vector comprises a coding sequence shown as SEQ ID No.1. Through establishment of the bifluorescence reporter vector p35mG with the coding sequence shown as the SEQ ID No.1, one fluorescent protein mCherry is taken as internal reference, the other fluorescent protein GFP is taken as a reporter gene, a to-be-detected gene segment is inserted into GFP coding sequence to form a recombinant reporter gene GFP-3'UTR, and the functional cis acting element in the RNA is rapidly identified by analyzing influence of different segments on fluorescence intensity ratio of GFP to mCherry; besides, the regulation process of the cis acting element can be controlled through application of different treatment or expression of different effector proteins, and accordingly, the regulation effect of different signals or effector proteins in plants on the RNA is studied.

An extensible RNA-based framework for engineering ligand-controlled gene regulatory systems, called ribozyme switches, that exhibit tunable regulation, design modularity, and target specificity is provided. These switch platforms typically contain a sensor domain, comprised of an aptamer sequence, and an actuator domain, comprised of a hammerhead ribozyme sequence. A variety of modes of standardized information transmission between these domains can be employed, and this application demonstrates a mechanism that allows for the reliable and modular assembly of functioning synthetic hammerhead ribozyme switches and regulation of ribozyme activity in response to various effectors. In some embodiments aptamer-regulated cis-acting hammerhead ribozymes are provided.

The invention discloses a lotus corniculatus AP2 / ERF transcription factor and an encoding gene and application thereof and belongs to the field of AP2 / ERF transcription factors and application. The invention discloses the gene of an AP2 / ERF transcription factor separated from lotus corniculatus. The nucleotide sequence of the gene is shown as SEQ ID NO.1. The amino acid sequence of protein encoded by the gene of the transcription factor is shown as SEQ ID NO.2. The invention further discloses an AP2 / ERF transcription factor structural domain which can be combined with a cis-acting element to start adverse-resistant response gene expression, and the amino acid sequence of the structural domain is shown as SEQ ID NO.3. Function conversion experiments prove that the gene of the AP2 / ERF transcription factor in a plant can improve salt tolerance of the transgenic plant remarkably, and it shows that the protein encoded by the gene of the transcription factor has a function of the AP2 / ERF transcription factor. The gene of the AP2 / ERF transcription factor has an important application prospect in improving resistance of the plant to adversity stress.

Disclosed are cis-acting regulatory elements from a KIM-1 gene. The elements can be used to direct the expression of operably linked sequences in renal tissue.

Plant expression construct are provided. According to an embodiment, the plant expression construct comprises a nucleic acid sequence encoding a hexokinase under a transcriptional control of a guard cell-specific cis-acting regulatory element. Also provided are methods of using the constructs and transgenic plants, plant cells and plant parts expressing same.

The invention discloses a soybean low-temperature inducing promoter, a recombinant expression vector containing the same, an application of the soybean low-temperature inducing promoter and particularly relates to the field of plant genetic engineering, in particular to expression of a soybean gene promoter sequence can serve as the promoter to control gene under low-temperature stress with an aim to effectively solve the problem that the soybean low-temperature inducing promoter is not developed yet. The promoter is derived from a promoter sequence of soybean ethylene response factor gene GmERF9, the promoter sequence is named as GmERF0P with a base sequence indicated as SEQ ID No: 1; the sequence contains multiple stress-related cis-acting elements; the cis-elements are respectively: GT-1, BIHD10S, WRKY, MYB, MYC and G-box. The soybean low-temperature inducing promoter is used for inducing cold-resistance gene expression under low temperature.

The invention discloses a newly found DNA sequence of the promoter of the chalcone synthase (chs) gene and the manufacturing method and use for the same. The promoter is used for PCR with a joint from sorbonne, and clones 899bp of the upstream regulation sequence of chs gene by combining the characteristics of the nested PCR, TD PCR, hot-started PCR and two-step PCR. The analysis in the promoter database Plant CARE shows that, the sequence has a basically conservative region TACPyAT which expresses specifically in flowers, and has the main cis-acting elements with which the primary promote is provided such as a transcription initiation related TATA box, a transcription auxiliary CAAT box, a G box and CCAAT etc. The promoter can regulate the specific expression of the target gene in the plant, and can provide an effective promoter element for the constructing the plant expression vector, increase the expression of foreign gene, and minimum the biological energy consumption.

The invention discloses a paeonia suffruticosa PsDREB1 gene anti-drought and high salinity promoter and its expression and an application, which belong to the technical field of plant gene engineering, and clones a promoter DREBP of Paeonia suffruticosa PsDREB1 gene. A nucleotide sequence of the prmoter contains 2245 bp base, the promoter area contains three ABRE cis acting elements, which can specifically perform response reaction to ABA; and the promoter area contains a MYB binding site related to drought induction. An expression vector pBI121 DERBP GUS constructed by gene promoter fragment can be used for strongly inducing the expression of a target gene when the plants are suffered with adversity stress such as drought and high salinity.