37 results about "TATA box" patented technology

An artificial promoter library (or a set of promoter sequences) for a selected organism or group of organisms is constructed as a mixture of double stranded DNA fragments, the sense strands of which comprise at least two consensus sequences of efficient promoters from said organism or group of organisms, or parts thereof comprising at least half of each, and surrounding intermediate nucleotide sequences (spacers) of variable length in which at least 7 nucleotides are selected randomly among the nucleobases A, T, C and G. The sense strands of the double stranded DNA fragments may also include a regulatory DNA sequence imparting a specific regulatory feature, such as activation by a change in the growth conditions, to the promoters of the library. Further, they may have a sequence comprising one or more recognition sites for restriction endonucleases added to one or both of their ends. The selected organism or group of organisms may be selected from prokaryotes and from eukaryotes; and in prokaryotes the consensus sequences to be retained most often will comprise the −35 signal (−35 to −30): TTGACA and the −10 signal (−12 to −7): TATAAT or parts of both comprising at least 3 conserved nucleotides of each, while in eukaryotes said consensus sequences should comprise a TATA box and at least one upstream activation sequence (UAS). Such artificial promoter libraries can be used, e.g., for optimizing the expression of specific genes in various selected organisms.

The invention relates to a grapevine powdery mildew resistance transcription factor gene VpRFP1 promoter sequence and application thereof. In the sequence, a full-length 1249bp promoter sequence of a Chinese wild vitis pseudoreticulata powdery mildew transcription factor gene VpRFP1 is cloned by adopting chromosomal walking in combination with a nested polymerase chain reaction (PCR) technology, wherein the sequence comprises 103 TATA-boxes and 27 CAAT-boxes, has 8 elements related to plant defense reaction, 14 photoreaction elements, 5 plant hormone response elements, and 14 other unknown elements. By adopting an agrobacterium tumefaciens-mediated instantaneously converted nicotiana benthamiana leaves analyzing method, the function of the Chinese wild vitis pseudoreticulata powdery mildew transcription factor gene VpRFP1 promoter proves that: the promoter has stronger promotion activity, can actively respond to pathogen invasion and disease resistant signal substances, and can improve the disease resistance and high temperature resistance of wheat, rice, potato, corn, soybean, rape and other plants through a transgenic method.

The present invention relates to cotton tissue specific and pathogenic bacterium inducing promoter and their application, and belongs to the field of biotechnology. The promoter contains CAAT-box, TATA-box, ethylene, methyl jasmonate, abscisic acid response element, pathogenic bacterium inducer response elements W boxes, GT-1, MYB, MYBST1, MYB1LEPR, etc. There are also specific root expressing elements to enhance the expression of GUS gene, which expresses specifically in the phloem of root, bud and stem, the vein of foliage and the guard cell of air pore. Paraffin section observation shows that GHNBS promoter expresses in phloem companion cell. After treatment with SA, MeJA, Ethylene, pathogenic bacteria of blight and pseudomonas syringae DC3000, GUS will have obviously raised activity, so that it is one organ specific and pathogenic bacterium relevant promoter.

There is provided an isolated polynucleotide sequence useful as a promoter in plant root tissue. The sequence is derived from the promoter for the Western white pine gene PR10. The sequence comprises at least 45 nucleotidcs, and has at least two Dof elements which individually may be in a forward or inverted orientation. Each Dof element has a sequence NNNWAAAGNNN, wherein N is A, T, G, or, C, and, W is A or T. The Dof elements are separated from each other by between 10 and 20 nucleotides. The sequence also has a TATA box located between 15 and 25 nucleotides in the 3′ direction of the closest Dof element. In some instances, the polynucleotide has one Dof element with the sequence NNNWAAAGNNN and another with the sequence NNNCTTTWNNN. In some instances the polynucleotide sequence is one where the 5′ Dof element sequence is CTCTCTTTAAT, and another Dof element (which may be the next Dof element) sequence is TTTTAAAGGTT and the TATA box sequence is TACAATAAATA or TATAWAWA, wherein W is A or T. Also provided are sequences useful in modulating gene expression in stem and leaf tissue.

A method for determining the risk of adverse effects of irinotecan (CPT-11), a synthetic anticancer drug, by detecting polymorphisms in the TATA box within the promoter region of the UDP-glucuronosyl transferase gene. A kit for detecting the adverse effects of irinotecan containing at least one pair of nucleic acid probes.

The invention provides bidirectional promoters for expressing two or more short RNA sequences from a single promoter. A particular embodiment of the bidirectional promoters of the invention include: 1) a Pol III promoter that contains a TATA box, a PSE and a DSE; and 2) a Pol III promoter that includes a PSE and a TATA box fused to the 5′ end of said DSE in reverse orientation. Vector embodiments are also disclosed comprising the novel bidirectional promoters of the invention, as well as methods of making and using these promoters.

The invention provides a promoter batch capture method. According to the promoter batch capture method, when a plant gene is transcribed, a complex of a 'DNA-protein factor' formed in a promoter area is utilized, an improved chromatin immunoprecipitation technology is applied, and DNA containing an unknown plant gene promoter fragment is sorted out. The promoter batch capture method is a technology for specifically capturing an unknown plant gene promoter fragment by utilizing the property that protein factors of OsTBP2 and OsTFIIB are tightly combined with promotion and transcribe of TATA box short-chain conserved sequence which on the promoter and through the generation of a specific antibody of the two factors and through the combination of the chromatin immunoprecipitation technology (ChIP). In the technology, by utilizing the advantages that most promoters contain one TATA box short-chain conserved sequence, and through the combination of technology that the unknown plant gene promoter fragment is separated through the newly developed chromatin immunoprecipitation technology (ChIP), large-scale capture of the plant promoter is easily achieved, and the technology has high commercial value.

The invention relates to a promoter for the specificity expression and the later development expression of a plant tissue and application thereof, belonging to the biotechnology field. In the invention, a promoter sequence of a cotton cyclopropane fatty acid synthase with the length of 2.6kb is obtained. Place analysis proves that the promoter sequence contains a CAAT box, a TATA box, a cupric ion induced expression element, a pathogenicbacteria induced expression element, a dry response element, a cold stress response element, and the like as well as some root specificity expression elements and floral organ specificity elements. After GUS (beta-Glucuronidase) is dyed, a traditional gene is specially expressed at a later development stage by the promoter, and the expression parts are only restricted at a stem base part, a root and an anther.

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 provides an inducible shRNA (short hairpin ribonucleic acid) lentiviral expression vector and a construction method and application thereof. The inducible shRNA lentiviral expression vector comprises an shRNA expression frame and a tetracycline repressor protein expression frame, wherein the shRNA expression frame comprises a first promoter, a tetracycline response element being combined with tetracycline repressor protein, a TATA box and an shRNA coding sequence, and the tetracycline repressor protein expression frame comprises a second promoter, a tetracycline repressor protein expression gene, an internal ribosome entry site (IRES) DNA (deoxyribonucleic acid) sequence and a marker gene. Compared with an existing inducible shRNA expression vector, the inducible shRNA lentiviral expression vector has the advantages of being capable of rapidly establishing inducible shRNA expressing a silent gene in various cell strains by using a single vector, having no need of clone selection of cells, and being applicable to in vitro culture and in vivo study of cells.

The invention provides an expression vector for expressing protein/polypeptide by a mammalian host cell expression protein/polypeptide. The expression vector contains a marker gene and a restriction-endonuclease enzyme digestion site, wherein the restriction-endonuclease enzyme digestion site is used for being inserted into insertion of a target gene; the quantity of Poly(A) tailing signals behindthe target gene is 2 or more than 2; a starting component of the marker gene is only a TATA box. The upstream end of the selectable marker gene of the expression vector provided by the invention contains two or more Poly(A) tailing signals; the interference, to the activity of the downstream TATA box, of an upstream strong promoter is minimized, so as to drive the expression of the selectable marker gene. The expression vector provided by the invention can be used for generating stable and further high-yield cell lines for the expression of recombinant protein within a short time; moreover, the gene amplification does not need drug selection/medication; the selection can be directly carried out in the condition that any no drug induced amplification does not exists, and these cell lines can be used for stably expressing high-level recombinant protein for a long time, and is quite applicable to the industrialized production of the protein or the polypeptide.

The invention provides a short small-nuclear RNA promoter as well as a construction method and application thereof. The length of the short small-nuclear RNA promoter is 150-300bp, and the short small-nuclear RNA promoter sequentially contains one TATA box conservative element, one USE conservative element and at least one MSP conservative element in the direction from an end 3' to an end 5'. Compared with an existing wild type small-nuclear RNA gene promoter, the short small-nuclear RNA promoter constructed in the invention has the advantages that the promoter has stronger transcriptional activity and obviously higher editing efficiency of driving the sgRNA and is shorter, the length of each sgRNA expression cassette can be reduced, the efficiency of constructing a vector by multi-targetsgRNA cloning is improved, the simultaneous editing of multiple targets and multiple genes is facilitated, and the promoter has a good application prospect in the aspect of genome editing.

The invention discloses an activated siRNA able to specifically enhance gene expression through a gene promoter with a TATA-box motif in the target. The activated siRNA has the characteristics that: (1) the activated siRNA is complementary with a position taking the TATA-box motif as the center; (2) the first two base groups of an antisense strand adopt UA; (3) the length is 23 nucleotides; (4) methylation modification is conducted on the 3' end of the antisense strand; and (5) mismatch of the 3' end of the antisense strand is avoided. The activated siRNA provided by the invention can significantly enhance the target expression for a long time, and has great application value and broad application prospects in biotechnology, gene therapy drug development and other fields.

The invention discloses a method for cloning the end of target gene 5'and a special kit thereof. The method comprises the following steps of: respectively combining a degenerate primer TDP which is designed according to a TATA-box conserved sequence with gene specific primers GSP1, GSP2 and GSP3 into 3 pairs of primers; and amplifying the end of the gene 5' by means of semi-nested PCR with the 3 pairs of primers, wherein the Tm values of the gene specific primers GSP1, GSP2 and GSP3 are higher than the Tm value of the TDP degenerate primer at 3-5 DEG C, and the combination positions between the gene specific primers GSP1, GSP2 and GSP3 and the gene are sequentially near to the end of the gene 5'. The method for cloning the end of target gene 5' does not need to use expensive RNA extraction reagent, RACE extraction reagent and the like, thereby reducing the experiment cost.

The invention discloses a nematode apoptosis regulatory gene ced-4 inducible promoter wn-2, a rice expression vector pCA-Wn-2-CED-4 and a preparation method of the pCA-Wn-2-CED-4 expression vector. According to the invention, a TATA-box core sequence shared by animal and plant genes is added into the reported core sequence having the best inductive effect in a W-box series of the promoter, therefore, a novel wn-2 sequence of a nematode apoptosis gene in rice expression is innovatively designed; the wn-2 and the inherent promoter of the rice expression vector are fused, so that the novel pCA-Wn-2-CED-4 expression vector of the rice expression vector is constructed; the pCA-Wn-2-CED-4 expression vector has the superior effect of inducing rice expression of the target gene ced-4; and the pCA-Wn-2-CED-4 expression vector is screened and connected to correct vector clone and successfully converted into rice varieties, such as Nipponbare and Lijiang Xintuan dark rice.