116results about How to "Increase transcriptional activity" patented technology

The present invention is directed to a DNA element that enhances the translation of the human amyloid precursor protein (APP) gene. The enhancer may be incorporated into expression vectors to enhance recombinant protein production. In addition, the invention is directed to an assay that utilizes vectors containing the translation enhancer element for the purpose of identifying agents that modulate the expression of the human amyloid precursor protein. These agents will ultimately be used to suppress APP expression in patients with Alzheimer's disease.

This invention describes the cloning and characterization of the promoter region for the PCA3dd3 gene. This region regulates the PCA3dd3 gene expression by a unique prostate specific transcriptional mechanism. The present invention relates to the use of this promoter region as a tool for prostate cancer treatment and diagnosis and screening of agents which regulate expression of PCA3. In a particular embodiment, the present invention relates to an isolated promoter sequence which comprises an isolated promoter sequence which comprises a sequence as set forth between nucleotide positions 372 to 460 of SEQ ID NO:1, this sequence enabling a prostate-specific modulation of transcription.

Disclosed is a new process for the production of recombinant proteins, by transient transfection of suspension-grown human embryonic kidney cells (293 cell line and its genetic variants) with an expression vector, using polyethylenimine (PEI) as a transfection reagent. In a preferred embodiment, the process uses 293E cells expressing the Epstein-Barr virus (EBV) EBNA 1 protein, in combination with an oriP-based episomal expression vector having an improved cytomegalovirus expression cassette comprising the CMV5 promoter. The process combines in a single step the cell growth, transfection and protein expression, is carried out without changing the culture medium, and allows to achieve high expression levels in a short period of time. The process may be carried out in a serum-free, low-protein culture medium, is easily scalable, compatible with continuous production processes, and fully adapted to high-throughput production of milligram quantities of recombinant proteins.

The invention relates to enhancer Hr3 adopting the nucleotide sequence shown in the SEQIDNO: 1. The enhancer Hr3 can be used for preparing and recombining heterologous protein by bombyx mori. The invention further relates to a recombination carrier containing the enhancer Hr3, and the recombination carrier is prepared in such a manner that the enhancer Hr3 is connected with a Nco1 restriction site at the upstream of a promoter of a carrier containing no enhancer through the Nco1 restriction site of the enhancer Hr3. In the invention, functional elements such as the first grade enhancer, activating transcription factor IE1, untranslated region sequences (5' UTR and 3' UTR) and the like which are expressed by intensifier are identified in the cellular level, and an efficient and stable sericin I expression system is built by integrating optimum elements on the basis, so that efficient expression recombination heterologous protein can be made of middle silk gland of transgenic bombyx mori.

The present invention features a method for determining the methyltransferase activity of a polypeptide and screening for modulators of methyltransferase activity, more particularly for modulators of the methylation of retinoblastoma by SMYD3. The invention further provides a method or pharmaceutical composition for prevention or treating of colorectal cancer, hepatocellular carcinoma, bladder cancer and / or breast cancer using a modulator so identified. N-terminal truncated forms of SMYD3 (alias ZNFN3A1) have higher methylating activity. Lys 824 is a preferred methylation site on the RB1 protein for SMYD3.

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 therapeutic and diagnositic methods for neoplasia treatment. The invention also relates to determining the prognosis of a neoplasia or determining a treatment protocal. Further features of the invention are methods for identifying NFAT target genes that promote neoplasia progression.

The invention belongs to the technical field of gene engineering, particularly relates to a rubber tree RNA (ribonucleic acid) polymerase type III promoter, in particular to a rubber tree U6 gene promoter proHbU6.6 and further discloses a cloning method and an application of the rubber tree U6 gene promoter proHbU6.6. The rubber tree RNA polymerase type III promoter, namely the rubber tree endogenous U6 promoter proHbU6.6 is obtained by cloning in a Para rubber tree for the first time; the promoter is a rubber tree endogenous RNA polymerase type III promoter, has high transcriptional activity,and can drive expression of downstream sgRNA (small guide ribonucleic acid); the activity of the promoter and feasibility of the promoter applied to a rubber tree CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / CRISPR-associated nuclease 9) gene editing system are demonstrated by instantaneous conversion of rubber tree protoplast; and CRISPR / Cas9 mediated rubber tree genometarget editing is achieved.

The invention relates to a novel guide RNA (Ribonucleic Acid) expression cassette used in a CRISPR / Cas (Clustered Regularly Interspaced Short Palindromic Repeats) system. According to the guide RNA expression cassette, a 5S rRNA gene is adopted as a starter to start expression of guide RNA. The invention further provides a CRISPR / Cas system with the guide RNA expression cassette and a method for editing genomes by using the system. The guide RNA expression cassette, the CRISPR / Cas system and the genome edition method provided by the invention have the advantages of universality, high efficiency, convenience, accuracy and the like.

Methods and compositions are provided for the diagnosis and treatment of heart diseases relating to cardiac hypertrophy, and for the regulation of proliferation and differentiation of cardiomyocyte progenitors in vitro. The detection of expression of components of the BAF complex, including, without limitation, detection of expression of Brg1, provides useful methods for early detection, diagnosis, staging, and monitoring of conditions leading to hypertrophy and enlargement of the heart. Manipulation of Brg1 activity provides for therapeutic intervention in the development of cardiac hypertrophy, where methods of decreasing Brg1 activity, e.g. through inhibition of binding, decreasing expression, and the like, reduces cardiac hypertrophy.

The invention discloses a myrothecium roridum A553 trichothecene synthase gene Tri5 promoter and application thereof. The nucleotide sequence of a promoter is shown in SEQ ID NO.1. According to the promoter, through a chromosome walking technology, the upstream promoter sequence of the Tri5 gene is obtained, the core region of the promoter is predicted, and the myrothecium roridum A553 trichothecene synthase gene Tri5 promoter Tri5P is obtained. By means of the promoter, expression of a hygromycin resistance gene can be efficiently started, the starting efficiency is close to that of a constitutive promoter TEF1, and a molecular biology basis is laid for increasing the yield of trichothecene toxin and obtaining more novel trichothecene toxin with high activities through transcriptional regulation and heterologous expression in the later period.

The present invention discloses a PPAR α / γ dual agonist and its application. The PPAR α / γ dual agonist comprises an effective amount of the compounds represented by formula I or / and its pharmaceutically acceptable derivative. Wherein, R1is selected from alkoxyl or ester group; R2 is selected from hydroxyl or ester group. The PPAR α / γ dual agonist according to the present invention can be used for preparing drugs and functional foods for preventing or / and treating metabolic syndrome, especially glucose or / and lipid disorders, with extensive and bright prospects of application.

The invention belongs to the technical field of gene engineering, and relates to a rubber tree RNA polymerase III type promoter, in particular to a rubber tree U6 gene promoter proHbU6.8. The invention further discloses a cloning method and application of the promoter. The rubber tree RNA polymerase III type promoter-rubber tree endogenous U6 promoter proHbU6.8 is obtained through cloning in a Brazil rubber tree for the first time. The promoter is the rubber tree endogenous RNA polymerase III-type promoter, the promoter has high-efficiency transcriptional activity, the expression of downstreamsgRNA can be driven, the activity of the promoter and the application feasibility of the promoter in a rubber tree CRISPR / Cas9 gene editing system are verified through transient transformation rubbertree protoplasm, and CRISPR / Cas9-mediated rubber tree genome targeted editing is achieved.

The invention relates to a corn ear length gene ZmEL1, a promoter, molecular markers linked to the promoter and applications of the gene and the molecular markers in screening ear length traits and cultivating long ear varieties, and belongs to the field of molecular genetics. The disclosed corn ear length gene ZmEL1 is characterized in that the nucleotide sequence of the gene is shown as any of SEQ ID NO. 2- SEQ ID NO.3; and an encoded amino acid sequence is shown as SEQ ID NO.1. Meanwhile, a promoter sequence (SEQ ID NO.4- SEQ ID NO.7), the molecular markers S2, S3 and S4 closely linked to the promoter, detection primer pairs of the markers (SEQ ID NO.8- SEQ ID NO.16) and a detection method are also disclosed. Further, a method for screening the corn ear length traits and increasing thelength of corn ears by utilizing the molecular markers is provided.

The invention discloses application of bavachinin and analogs of bavachinin. According to the application, bavachinin or / and analogs of bavachinin serve as active ingredients to prepare compositions for enhancing exciting activity of PPARalpha or / and PPARgamma. Research results show that bavachinin and analogs of bavachinin can remarkably improve the transcriptional activity of a PPARgamma exciting agent to PPARgamma, enhance the transcriptional activity of a PPARalpha exciting agent to PPARalpha and serve as active ingredients of PPARalpha or / and PPARgamma exciting agents to prepare compositions for preventing or treating the metabolic syndrome, the application range of bavachinin and analogs of bavachinin can be widened, and decrement and synergism of PARalpha or / and PPARgamma exciting agents can be achieved to reduce the toxic and side effects of PARalpha or / and PPARgamma exciting agents and achieve wide clinical application of PARalpha or / and PPARgamma exciting agents.

The invention discloses a DNA fragment with a promoter function and a use thereof. The DNA fragment has a sequence selected from (a), a nucleotide sequence shown in the formula of SEQ ID NO. 1 or its complementary sequence, (b), a nucleotide sequence which is derived from the nucleotide sequence shown in the formula of SEQ ID NO. 1 through replacement, deletion or addition of one or multiple nucleotides and has a promoter function the same to that of the nucleotide sequence shown in the formula of SEQ ID NO. 1, or its complementary sequence, and (c), a sequence derived from the nucleotide sequence shown in the formula of SEQ ID NO. 1 through addition of one or more ribosome binding sites. The DNA fragment has a promoter function and strong specific expression activity, realizes high exogenous gene expression without an inducer and provides an effective element for bacillus subtilis expression of an exogenous gene.

The invention provides a novel method for generating stable mammalian cell lines with enhanced protein production capabilities, and to expression vectors and related methods for high level expression of biopharmaceutical proteins of interest.

The invention relates to a chimeric promoter construct useful for neuronal cell specific gene expression. The invention provides a recombinant nucleic acid molecule that comprises a neuronal cell specific promoter such as platelet-derived growth factor β-chain promoter, operably linked to a heterologous enhancer which enhances the transcriptional activity of the promoter, such as cytomegalovirus immediate early gene enhancer. The promoter construct according to the invention can increase and prolong gene expression in neuronal cells and may be advantageously used in gene therapy of neuronal disorders.

The invention belongs to the technical field of gene engineering, particularly relates to a rubber tree RNA polymerase III-type promoter, more specifically relates to a rubber tree U6 gene promoter proHbU6.3, and further discloses a cloning method and applications thereof. The present invention clones and obtains the rubber tree RNA polymerase III-type promoter-rubber tree endogenous U6 promoter proHbU6.3 in hevea brasiliensis for the first time, the promoter is rubber tree endogenous RNA polymerase III-type promoter, the promoter has high-efficiency transcription activity and can drive downstream sgRNA expression, the activity and the feasibility of the promoter for application in rubber tree CRISPR / Cas9 gene editing system are verified by transient transformation of rubber tree protoplasts, and CRISPR / Cas9 mediated rubber tree genome targeted editing is realized.

The present inventors successfully constructed expression vectors that enable high-level production of foreign gene-derived proteins in mammalian host cells, which comprise a translation-impaired drug resistance gene cistron whose expression has been attenuated by altering the codons to the least frequently used codons in mammals; and a gene cassette which has a cloning site for incorporation of a foreign gene between a highly transcriptionally active promoter and a highly stable polyadenylation signal.

The invention discloses a promoter of a trichothecene synthase gene Tri12 of Leucosphaeria dothidea A553 and the application of the promoter. The nucleotide sequence of the promoter is as shown in SEQID NO. 1. According to the invention, an upstream promoter sequence of the Tri12 gene is obtained through a chromosome walking technology, and a promoter core region is predicted so as to obtain thepromoter Tri12P of the trichothecene synthase gene Tri12 of the Leuconostoc dunalianum A553. The promoter can efficiently start expression of the hygromycin resistance gene hph, and the starting efficiency is similar to the starting efficiency of a constitutive promoter TEF1, so a molecular biological foundation is laid for increasing the yield of trichothecene toxin and obtaining more high-activity novel trichothecene toxin through transcription regulation and heterologous expression in a later period.

The present invention identified OVARC1000473 (SEQ ID NO: 1) and NT2RM1000377 (SEQ ID NO: 3) as clones showing suppression of CREB activation by forskolin, and provides evaluation methods using these genes, and / or proteins encoded by these genes. Furthermore, these proteins were found to enhance cell damage. Compounds that can be screened based on the evaluation methods of this invention are useful as agents for inhibiting the CREB dephosphorylation reaction, agents for suppressing enhancement of cell damage, and preventive and therapeutic agents for memory disorders and / or neurodegenerative disorders.

The invention discloses a pulmonary hypertension virulence gene ACVRL1 mutation site and an application thereof. The amino acid sequence of a protein provided by the invention is sequence 4. The DNA molecule of the protein is coded and the nucleotide sequence of the DNA molecule is sequence 2. The experiment shows that the novel ACVRL1 gene mutation site is related to the pulmonary hypertension, by means of detecting whether the novel gene mutation site is the mutation site, whether the sample to be tested suffers from pulmonary hypertension can be predicted; and the novel gene mutation site can be used as a target for treating the pulmonary hypertension, and a new therapy pathway for researching the pulmonary hypertension drugs can be provided. The cell experiment verifies that the ACVRL1 gene mutation site is capable of reducing the SMAD1 and SMAD5 protein phosphorylation and / or increasing the BMP transcriptional activity in the cell.

The invention belongs to the technical field of severe asthma action mechanisms, and discloses a medicine for relieving severe asthma, application and an animal model construction method, the medicine is an HDAC10 inhibitor, and the action mechanism of HDAC10 for regulating and controlling Th17 / IL-17A expression in severe asthma is clarified by using the construction method; the interaction mode of HDAC10 and STAT3 is defined; it is proved that HDAC10 deacetylated STAT3 regulates and controls the expression of Th17 / IL-17A in severe asthma. it is determined that the HDAC10 inhibitor can relieve severe asthma, and an experimental basis is provided for HDAC10 targeted therapy of severe asthma. it is determined that the HDAC10 inhibitor can relieve severe asthma, and an experimental basis is provided for HDAC10 targeted therapy of severe asthma. Compared with a control group, the intraperitoneal injection of the HDAC10 inhibitor can improve airway inflammation, airway hyperreactivity and airway mucus secretion of a severe asthma mouse model mainly comprising neutrophile granulocytes.

The invention discloses a promoter. The length of the promoter is 150-1000bp, wherein the promoter comprises a nucleic acid fragment expressed as SEQ ID NO.1; the promoter is capable of starting the transcription in bacillus subtilis. The invention also provides a recombinant expression vector; a reading frame is allowed to be inserted into the recombinant expression vector and is used for expressing protein in bacteria; a promoter of the recombinant expression vector comprises the promoter. The invention further provides a method for expressing heterologous proteins. The method comprises a step of inserting the reading frame for encoding the heterologous proteins into the recombinant expression vector to prepare recombinant expression plasmids. The invention also provides application of the promoter to preparation of the recombinant expression vector and / or expressing the proteins in the plasmids. By virtue of the technical scheme, the expression quantity of the heterologous proteins can be obviously increased without induction.

The invention provides a method for simultaneously activating multiple porcine endogenous stem cell factors by adopting a tandem sgRNA strategy. Blasticidin and hygromycin drugs are adopted for screening and constructing a PK-15 cell line (PK-15-dCas9-MS2) capable of stably expressing dCAS-VP64 and MS2-P65-HSF1 first, sgRNAs capable of targeting promoters of the porcine stem cell factors Oct4, Sox2, Klf4, c-Myc and miR-302 / 367 are designed and constructed separately, high-activity sgRNA screening is carried out on the cells, the five sgRNAs with the highest gene transcription activation efficiency are serially combined and constructed into an expression vector, and the expression vector is introduced into the PK-15-dCas9-MS2 cells to simultaneously activate the transcriptional activity ofporcine OSKM and miR-302 / 367. The method has a potential application value in induced generation of porcine iPSC.

The invention discloses a benzisothiazole hypoxia-inducible factor 2 agonist compound or a pharmaceutically acceptable salt thereof, the compound can stimulate the transcriptional activity of a hypoxia-inducible factor 2 and enhance the generation and secretion of erythropoietin so as to promote the generation of erythrocytes; the prepared benzisothiazole hypoxia-inducible factor 2 agonist compound or the pharmaceutically acceptable salt thereof can be combined with a prolyl hydroxylase inhibitor to play a synergistic role in improving the transcriptional activity of HIF-2, and can be used for treating hypoxia-inducible factor 2 related diseases, such as ischemic diseases and the like.

The invention discloses a compound with PPAR multiple agonist activities and a preparation method and application thereof. The compound has the chemical structure as shown in the formula I. Research results show that the compound adopting the formula I can remarkably improve the transcriptional activities of PPAR alpha, beta and gamma, has the multiple agonist activities of PPAR alpha, beta band gamma, is expected to serve as an active ingredient to be used for preparing a drug or health food for preventing and / or treating metabolic syndromes, and is particularly expected to be used for preparing a drug or health food for preventing and / or treating abnormal glucose metabolism and / or abnormal lipids metabolism diseases. The compound has a wide application prospect and a remarkable medicinal value.