421 results about "Promoter activity" patented technology

Isolated polynucleotide sequences exhibiting endothelial cell specific promoter activity, novel cis regulatory elements and methods of use thereof enabling treatment of diseases characterized by aberrant neovascularization or cell growth are disclosed.

Isolated polynucleotide sequences exhibiting endothelial cell specific promoter activity, novel cis regulatory elements and methods of use thereof enabling treatment of diseases characterized by aberrant neovascularization or cell growth are disclosed.

Isolated polynucleotide sequences exhibiting endothelial cell specific promoter activity, novel cis regulatory elements and methods of use thereof enabling treatment of diseases characterized by aberrant neovascularization or cell growth are disclosed.

A variety of methods and compostions are provided, including methods and compositions for targeted modification of a specific target site in a cell or organism, methods for integrating polynucleotides of interest, methods to assess promoter activity, directly select transformed organisms, minimize or eliminate expression resulting from random integration into the genome of an organism, such as a plant, remove polynucleotides of interest, combine multiple transfer cassettes, invert or excise a polynucleotide, silence a gene, and identify and / or characterize transcriptional regulating regions. The methods involve the introduction of a cell proliferation factor and a double-strand break-inducing enzyme into an organism.

The present invention relates to methods of identifying candidate compounds for the treatment of leukemia and diagnostic methods based on histone methylation and HoxA5 promoter activity.

Isolated polynucleotide sequences exhibiting endothelial cell specific promoter activity, novel cis regulatory elements and methods of use thereof enabling treatment of diseases characterized by aberrant neovascularization or cell growth are disclosed.

The present invention provides human involucrin (hINV) sequences having tissue specific and cell type specific promoter activity. The sequences provided herein direct expression to suprabasal cells of stratifying epithelia. The invention further provides methods for the production of transgenic animals which contain a hINV promoter sequence which directs the expression of human papillomavirus 16 oncogenes (or other oncogenes). These animals display cervical and epidermal hyperplasias as well as cancer of the trachea, esophagus, colon, epidermis, anus / rectum, lymph nodes, spleen and lung. The animals of the invention provide a useful model for screening potential anti-neoplastic compounds, carcinogens, and co-carcinogens for a number of cancers.

A method for analyzing the temporal behavior of gene expression for a group of genes which are part of a biological system or subsystem. Preferably, such an analysis enables the order of expression of such genes to be determined. More preferably, the temporal behavior of gene expression is assessed according to the analysis of the kinetics of gene transcription. According to a preferred embodiment of the present invention, the kinetics of gene transcription are measured according to promoter activity of a plurality of genes. More preferably, such kinetics are measured in a living organism or a portion of such an organism, such as a cell for example. For single-celled organisms, such as bacteria for example, the kinetics may easily be measured for the entirety of the living organism.

The invention discloses a method for detecting promoter activity by utilizing double luciferase reporter genes. The method for detecting the promoter activity by utilizing the double luciferase reporter genes comprises step 1, building a pGL3-basic-MSTNpro recombinant which contains 7 sections of MSTN gene 5' control region fragments with different lengths; step 2, performing cultivation and planking on target cells, configuring a mixture of the pGL3-basic-MSTNpro of the step 1 and a lipidosome and enabling a renilla luciferase carrier pRL-TK to be served as an internal reference to perform cell co-transfection; step 3, performing detection on luciferase activity through the double luciferase reporter genes.

A method of titering adeno-associated virus particles in a sample, said method comprising the steps of contacting target cells with a DNA synthesis inhibitor and an agent that increases the activity of the CMV immediate early promoter; contacting target cells treated as in step (a) with a sample containing adeno-associated virus particles; and determining the number of target cells infected by said adeno-associated virus particles in said sample, wherein said number of target cells infected is directly proportional to the titer of said particles in said sample, thereby determining the titer if said adeno-associated virus particles.

The present invention relates to methods for increasing the secretion of a protein of interest (POI) from a eukaryotic cell comprising co-expression of a POI and of at least one protein that enhancesprotein secretion, said enhancing protein being selected from the group consisting of BMH2, BFR2, C0G6, C0Y1, CUP5, IMH 1, KIN2, SEC31, SSA4 and SSE1. The invention further relates to a yeast promotersequence, in particular to a promoter sequence of the PET9 gene of P. pastoris, having, under comparable conditions, an increased promoter activity relative to a promoter sequence of the GAP protein.The invention further relates to an expression vector comprising such a promoter sequence and to the use of such an expression vector for expression of a POI in a host cell. The invention further relates to new yeast promoter sequences of genes from P. pastoris, which are useful for expression of a POI in yeast.

The invention discloses a promoter with a nucleotide sequence as follows: 1) a nucleotide sequence shown in sequence 1 in a sequence table; 2) a nucleotide sequence which hybridizes with the nucleotide sequence in 1) under strict conditions and has promoter activity; or 3) a nucleotide sequence which has homology of more than 70% with the nucleotide sequence in 1) and has promoter activity. The promoter provided by the invention can be applied to the biotechnology field with corynebacterium or escherichia bacteria as industrial microbes. Experiments prove a fragment with promoter activity from SEQ ID NO:1 has the activity higher than that of a trc promoter (SEQ ID NO:2).

A method for treating woodchips, comprising the steps of: preparing a DNA encoding an antisense RNA substantially complementary to the whole or a part of a transcription product of a cellulolytic enzyme gene derived from Basidiomycete; preparing a vector comprising (a) the above DNA, or (b) a recombinant DNA comprising the above DNA and a DNA fragment having a promoter activity, wherein the above DNA binds to the above DNA fragment such that an antisense RNA of the cellulolytic enzyme gene is generated as a result of transcription; transforming host cells with the above vector, so as to prepare the host cells having a suppressed cellulolytic enzyme activity; and inoculating the above host cells having a suppressed cellulolytic enzyme activity into woodchips to treat them.

The present invention relates generally to constructs and their use in gene expression or gene regulation assays. More particularly, the present invention provides expression vectors and / or reporter vectors providing kinetics of protein expression with improved temporal correlation to promoter activity. Even more particularly, the invention provides expression vectors comprising a transcribable polynucleotide which comprises a sequence of nucleotides encoding a RNA element that modulates the stability of a transcript corresponding to the transcribable polynucleotide. The present invention provides, inter alia, novel vectors, useful for identifying and analysing cis- and trans-acting regulatory sequences / factors as well as vectors and genetically modified cell lines or organisms that are particularly useful for drug screening and drug discovery.

A promoter domain of 1816 bp or 441 bp in the upstream side of exon 1B of IAI.3B gene has a specifically high promoter activity in ovarian cancer cells. An adenovirus having this promoter domain inserted in the E1 domain thereof exhibits a specifically high cell proliferation inhibitory effect on ovarian cancer cells. Thus, it is efficacious in gene therapy for ovarian cancer.

Provided are native promoters comprising polynucleotides isolated from Corynebacterium glutamicum, and mutant promoters derived therefrom, which may be used to regulate, i.e., either increase or decrease, gene expression. Also provided are promoter ladders comprising a plurality of the promoters having incrementally increasing promoter activity. Also provided are host cells and recombinant vectors comprising the promoters, and methods of expressing genes of interest and producing biomolecules using the host cells.

The invention discloses a DNA (Deoxyribose Nucleic Acid) with constitutive promoter activity, application of the DNA and a pichia pastoris expression vector. The DNA has a base sequence as shown in SEQ No.1 (Sequence Number); and the application of the DNA relates to the application of the DNA in construction of the pichia pastoris (Pinchia Pastoris) expression vector. The DNA disclosed by the invention has the constitutive promoter activity, and can activate the transcription of a downstream structural gene without an inductor; the transcriptional activity shows little change in four different culture mediums, namely, ethanol, methanol, glucose and glycerol; the promoter activity is efficient, and the efficiency of the initiation transcription is four times more than the pichia pastoris GAPDH (Reduced Glyceraldehyde-phosphate Dehydrogenase) promoter. The pichia pastoris expression vector, constructed by the DNA disclosed by the invention, can efficiently express the extrinsic protein without methanol induction, and the efficiency of expressing the extrinsic protein (Enhanced Green Fluorescent Protein) is about 6 to 8 times that of the expression system of the GAPDH promoter and 1.5 to 2 times that of the expression system of a TEF1 (Transcription Enhancer Factor 1) promoter.

The present invention provides a nucleic acid molecule having enhanced promoter activity, which is operably linked to a gene encoding aspartate aminotransferase and derived from Corynebacterium glutamicum, a vector comprising the nucleic acid molecule, a transformant transformed with the vector, and a method for producing L-lysine using the transformant.

According to the present invention, a DNA construct of interest is assembled from overlapping subfragments via an acceptor module which comprises the distal end of the construct at a position downstream from a promoter. The construct is assembled distal to proximal via homologous recombination events occurring in the span between that distal end of the construct and the upstream end of the promoter. These recombination events occur iteratively between the acceptor module and alternative donor modules. Successful recombination places one of at least two marker genes under the transcriptional control of an active form of the promoter. As a result of alternating use of two varieties of donor modules, as few as two selection markers may be used to produce a complex DNA construct.

The invention provides a Gluconobacter oxydans (G.oxydans) promoter and its application. The promoter has the following nucleotide sequence: 1) a nucleotide sequence shown as SEQ ID NO:1 in a sequence table; or 2) a nucleotide sequence hybridizing with the nucleotide sequence shown as SEQ ID NO:1 under rigorous conditions and having promoter functions; or 3) a nucleotide sequence which is obtained by subjecting the nucleotide sequence defined by 1) or 2) to substitution, deletion, insertion or adding by one or more basic groups, and has over 90% homology with the defined nucleotide sequence and promoter functions. The promoter provided by the invention has the function of effective expression of the endogenous or exogenous gene in Gluconobacter oxydans, and has higher promoter activity compared with common promoters. The Gluconobacter oxydans promoter can realize application to gene expression and functional gene screening research in G.oxydans, and also can be applied to construction of 2-KGA high-yield gene engineering bacteria.

The invention discloses a T carrier that can directly and directionally clone a promoter and research the activity of the promoter and a construction method thereof. The construction method comprises the steps: firstly: an anterior T carrier with a Xcml box (or an AhdI box) is prepared, and the anterior T carrier comprises a DNA spacer sequence that has different resistance genes or other selective markers with the initiating carrier and two Xcml enzyme cutting site sequences that are closely connected with two sides of the DNA spacer sequence respectively; secondly, the T carrier is obtained by enzyme cutting the anterior T carrier in the first step with the Xcml enzyme or isoschizomer of the Xcml enzyme; the Xcml box in the first step is obtained by amplifying PCR, and the recognition site of the upstream primer Xcml enzyme still contains the first half part of the recognition site of the rare restriction enzyme Pmel which is used for the directional cloning analysis of the promoter PCR products. The method in the invention is also applicable to the preparation of the T carrier of a common cloning type and an expressive T carrier, and plays an important role in the genetic engineering field.

This invention is related to the extraction of a protein from the seeds, cotyledons or plantlets of Lupinus genus, as well as to the way of producing it in recombinant form and of expressing it in genetically modified plants. Due to the exceptional characteristics exhibited by this protein in what concerns: its potent antifungal and anti-Oomycete activity, which confers great potential to the protein as a fungicide, (2) its strong plant growth promoter activity, particularly notorious on unhealthy or naturally weakened plants, (3) its extreme resistance to denaturation, which allows the use of the protein under field conditions, (4) its great susceptibility to proteolytic attack, which makes it harmless to the environment and nontoxic for man, and (5) its well balanced amino acid composition. It is claimed its use, or of any modification of the protein that maintains its biological properties, as a supplement in human or animal nutrition and as a fungicide, insecticide, growth promoter, fertilizer or in the preparation of genetically modified organisms.

The invention belongs to the technical field of animal gene engineering and particularly relates to isolated identification and functional verification of different-length promoter regions of pig skeletal muscle specificity expression genes alpha-actin. Eight upstream different-length promoters of the skeletal muscle specificity expression genes alpha-actin (the Gene Bank accession number is 100154254) are cloned from the pig genome, and the nucleotide sequences of the promoters are respectively shown as SEQ ID No: 2 to SEQ ID No: 9. The results show that the region with the length being 249bp has independent promoting activity and muscle tissue specificity, the nucleotide sequence is shown as the SEQ ID No: 9 in a sequence table. The invention also discloses a method for obtaining eight different deletion promoter segments, a method for preparing corresponding recombinant expression vectors and an application of a dual-luciferase enzymatic activity detection system to the promoter activity analysis.

Provided are native promoters comprising polynucleotides isolated from Corynebacterium glutamicum, and mutant promoters derived therefrom, which may be used to regulate, i.e., either increase or decrease, on-pathway and / or off-pathway gene expression. Also provided are promoter ladders comprising a plurality of the promoters having incrementally increasing promoter activity. Also provided are hostcells and recombinant vectors comprising the promoters, and methods of expressing ancillary genes of interest and producing biomolecules using the host cells.

The invention belongs to the field of animal gene engineering. Seven promoters Six1-P, P1, P2, P3, P4, P5 and P6 with different lengths in the upstream of a specific expression gene of skeletal muscles in pigs are cloned from a pig genome. Nucleotide sequences of the promoters are shown in SEQ ID NO:1; 2; 3; 4; 5; 6; and 7. The lengths of the nucleotide sequences are respectively 1,692, 180, 530, 844, 1,143, 1,135 and 1,494bp. The activity analysis on promoter fragments with different lengths of a pig Six1 gene in three eukaryotic cells shows that the promoters of the pig Six1 gene are promoters which express the muscle specificity; except for the P1, other promoters P2, P3, P4, P5 and P6 all have the capacity of promoting gene expression; and in the promoters P2, P3, P4, P5 and P6, except for the promoter P2, other promoters all show the muscle specificity. The invention discloses the seven promoters, a method for preparing corresponding expression vectors of the seven promoters and identification of the promoter activity of the pig Six1 gene by using a dual-luciferase reporting system.

The invention relates to a high-throughput screening system constructed by taking I type collagen alpha1 gene COL1A1 promoter activity as a target spot on the basis of the characteristic that I type collagen alpha1 is highly expressed in a hepatic fibrosis process. By constructing an I type collagen alpha1 gene COL1A1 promoter fragment and pGL4.17 expression vector recombinant plasmid, transfecting a human hepatic satellite cell line (LX2) and screening by taking COL1A1 promoter activity detected by virtue of luciferase expression strength of a carrier pGL4.17 as index to obtain a stable monoclonal cell strain LX2-COL, and screening compounds S1-S7 by applying the cell strain LX2-COL to obtain a compound S7 with an obvious inhibiting effect on the COL1A1 promoter activity. mRNA real-time PCR and Western blotting analysis respectively show that the compound S7 can obviously reduce expression of the I type collagen alpha1 in transcription and protein levels and is hopeful to become a candidate anti-hepatic fibrosis drug; and the mRNA real-time PCR and Western blotting analysis also respectively show that the constructed system can be applied to anti-hepatic fibrosis drug high-throughput screening.

The invention discloses a plant induction type promoter and application of the plant induction type promoter. The nucleotide sequence of the promoter is the nucleotide sequence shown by SEQ ID No.1 in a sequence table. Experiments prove that under the conditions of different concentrations and different processing time of NaCl, PEG and ABA, the promoter HbHAK2 can drive the GUS gene expression; and under the non-processed (comparison), low temperature (4 DEG C) and high temperature (42 DEG C) stress, the HbHAK2 cannot drive the GUS gene expression. The result shows that the HbHAK2-744bp promoter is a novel induction type promoter; and high promoter activity can be realized under the salt or drought stress or the ABA induction; and the target gene expression can be inducted for adapting to the environment. The promoter can be applied to stress-tolerance gene engineering vector construction, plant stress-tolerance molecular mechanism study and crop salt-resistant and drought-resistant transgenic breeding. An economic fast and effective path is provided for plant genetic modification.

The invention relates to a cotton GhFP1 gene which belongs to a member of a bHLH transcription factor gene family, the cDNA (complementary deoxyribonucleic acid) full length is 1262bp, and the cotton GhFP1 gene can code a bHLH protein containing 355 amino acids. GhFP1 is respectively expressed in various tissues of cotton, and is highly expressed in the fiber development premetaphase. The GhFP1 promoter segment with the length of 1061bp is separated and cloned. The analysis on the activity of the promoter by using the GUS report gene indicates that the promoter has higher specific expression activity in cotton fibers. The experiment indicates that the overexpression of GhFP1 in Arabidopsis thaliana results in increase of leaf epidermal hair; and the overexpression of GhFP1 in cotton results in enhancement of expression of related genes of cotton fibrocyte brassinolide and increase of ripe cotton fiber length. The result proves that the GhFP1 protein performs positive regulating actions and promotes elongation of fibrocytes in the cotton fiber elongation process, and thus, possibly has important application value in improving the cotton fiber quality and increasing the cotton yield.

The invention discloses Brassica juncea disease-resistant protein BjMYB9 as well as an encoding gene sequence and application thereof. The amino acid sequence of the disease-resistant protein BjMYB9 is as shown in SEQ ID NO: 1, and the nucleotide sequence of the encoding gene of the disease-resistant protein BjMYB9 is as shown in SEQ ID NO: 2. A R2R3-MYB type transcription factor capable of being specifically bound with a fungus inducing core element W-box-like-4 in a promoter BjC-P of a Brassica juncea disease-resistant gene BjCHI1 is screened with a yeast one-hybrid technology and is named as BjMYB9. A gel retardation experiment and a tobacco transient expression experiment prove that BjMYB9 can be specifically bound with W-box-like-4 and activates the promoter. Expression of the BjMYB9 gene in Brassica juncea RNA is obviously induced by fungi, the gene can significantly enhance the resistance of arabidopsis thaliana to Botrytis cinerea after overexpression in a model plant arabidopsis thaliana, thereby providing an experimental basis and genetic resource for revealing of the plant disease-resistant mechanism and cultivation of disease-resistant varieties.