36 results about "TATA box" patented technology

The invention relates to an AaPDR2 gene promoter as well as a functional verification method and application thereof. The AaPDR2 gene promoter comprises the following cis-acting elements: ABRE, BoxI, CAAT-box, CCAAT-box, CGTCA-motif, G-Box, G-box, GA-motif and TATA-box. Further, the DNA (Deoxyribonucleic Acid) sequence of the AaPDR2 gene promoter is shown in SEQ ID NO.1. The AaPDR2 gene promoter has a function of predominantly expressing genes guided by the AaPDR2 gene promoter in glandular secretory trichome and T shape trichome of young leaves of Artemisia annua L., and the function is confirmed by a GUS reporter gene. Therefore, the AaPDR2 gene promoter disclosed by the invention has important significance for genetic engineering breeding for expressing and generating metabolites by using plant glandular trichome tissues.

An allergen-free transgenic peanut seed is produced by recombinant methods. Peanut plants are transformed with multiple copies of each of the allergen genes, or fragments thereof, to suppress gene expression and allergen protein production. Alternatively, peanut plants are transformed with peanut allergen antisense genes introduced into the peanut genome as antisense fragments, sense fragments, or combinations of both antisense and sense fragments. Peanut transgenes are under the control of the 35S promoter, or the promoter of the Ara h2 gene to produce antisense RNAs, sense RNAs, and double-stranded RNAs for suppressing allergen protein production in peanut plants. A full length genomic clone for allergen Ara h2 is isolated and sequenced. The ORF is 622 nucleotides long. The predicted encoded protein is 207 amino acids long and includes a putative transit peptide of 21 residues. One polyadenilation signal is identified at position 951. Six additional stop codons are observed. A promoter region was revealed containing a putative TATA box located at position −72. Homologous regions were identified between Ara h2, h6, and h7, and between Ara h3 and h4, and between Ara h1P41B and Ara h1P17. The homologous regions will be used for the screening of peanut genomic library to isolate all peanut allergen genes and for down-regulation and silencing of multiple peanut allergen genes.

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.

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 present invention provides an expression vector for expressing proteins / polypeptides in mammalian host cells, comprising a marker gene and a restriction endonuclease cleavage site for inserting a target gene, and a Poly(A) tail after the target gene The number of signals is two or more, and the promoter element of the marker gene is only the TATA box. The upstream of the selectable marker gene of the expression vector provided by the present invention contains two or more Poly(A) tailing signals to minimize the interference of the strong upstream promoter on the activity of the downstream TATA box to drive the expression of the selectable marker gene. The expression vector of the present invention can generate stable and high-yielding cell lines for recombinant protein expression in a short time, and gene amplification does not require drug selection / mediation, and can be directly selected without any drug-induced amplification, These cell lines can stably express high levels of recombinant proteins for a long time and are very suitable for industrial production of proteins or polypeptides.

The invention provides a method for capturing promoters in batches. The method uses the complex of "DNA-protein factors" formed in the promoter region when plant genes are transcribed, and applies the improved chromosomal immunoprecipitation technique to capture the unknown plant DNA sorting out of gene promoter fragments. The present invention utilizes the property that the protein factors OsTBP2 and OsTFIIB tightly bind to the TATA box short-chain conserved sequence on the promoter to initiate transcription, and generates specific antibodies for these two factors, combined with chromatin immunoprecipitation (ChIP), to specifically target A technique for capturing promoter fragments of unknown plant genes. This technology utilizes the characteristic of a short-chain conserved sequence of the TATA box contained in most promoters, combined with the newly developed chromatin immunoprecipitation technique (ChIP) in recent years to isolate unknown plant gene promoter fragments, it is relatively easy Achieving large-scale capture of plant promoters has high commercial value.

The present invention provides an RNA polymerase III promoter characterized in that a part of its sequence is substituted with a sequence capable of undergoing specific DNA recombination by Cre recombinase in such a manner functions of PSE and a TATA box are not impaired. This RNA polymerase III promoter can be used in a RNAi induction method with the condition that RNAi molecules to be used in analyzing gene function can be induced at an arbitrary point of time or only in a specific tissue (organ).

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.

The invention relates to an AaPDR2 gene promoter as well as a functional verification method and application thereof. The AaPDR2 gene promoter comprises the following cis-acting elements: ABRE, BoxI, CAAT-box, CCAAT-box, CGTCA-motif, G-Box, G-box, GA-motif and TATA-box. Further, the DNA (Deoxyribonucleic Acid) sequence of the AaPDR2 gene promoter is shown in SEQ ID NO.1. The AaPDR2 gene promoter has a function of predominantly expressing genes guided by the AaPDR2 gene promoter in glandular secretory trichome and T shape trichome of young leaves of Artemisia annua L., and the function is confirmed by a GUS reporter gene. Therefore, the AaPDR2 gene promoter disclosed by the invention has important significance for genetic engineering breeding for expressing and generating metabolites by using plant glandular trichome tissues.

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 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 present invention relates to small secreted reporter-peptides 15 to 150 amino-acids in length comprising a response element, activated by one or more transcription factors induced by the pharmacology active substance to be analyzed following its interaction with a specific intracellular or cell surface molecule, functionally linked to a response element, a TATA box, a signal peptide, anchor and detection sequences to which antibodies can be raised, and a poy-A tail. The anchor sequence may differ from one peptide to another or may be common to multiple secreted reporter-peptides such that all the peptides can be analyzed simultaneously by ELISA using a detection antibody labelled with for example HRP, or by the use of a commonly available immuno-detection platform such MSD, Gyros, AlphaLisa, or Biacore. The detection sequence is unique to each secreted reporter-peptide and may be labelled with a Sulfo-Tag that permits detection of the peptide on the MSD platform, or Alexa that permits detection on the Gyros platform, or digoxigenin that permits detection of on the PerkinElmer AlphaLISA platform, or left unlabeled for detection by SPR on a Biacore platform. The present invention provides i.a. a substantial improvement of cell-based assays for analysis using automated immune-detection platforms and allows simultaneous analysis of multiple analytes, multiple sampling from a single cell culture, and obviates the necessity to lyse cells and remove cell debris by centrifugation prior to analysis on an automated immuno-detection platform. The present invention also provides a means of increasing the dynamic range, sensitivity and reducing the cost of cell-based assays and can be applied to existing engineered cell lines, such as those containing a reporter- gene such as a luciferase reporter-gene obviating the necessity to extensively re-engineer cell lines containing multiple molecular constructs.