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  • antibiotic resistance
  • Following λ-red operon activity induction, a linear, double-stranded cassette encoding a selectable marker, such as antibiotic resistance, is transformed into the cells in place of the target gene and incorporated into the DNA behind a specific inducible promoter. (wikipedia.org)
  • A plasmid expressing flippase (FLP) can be transformed into the recombined cells, which can specifically cleave FLP recognition target sites (FRTs) flanking the antibiotic resistance gene. (wikipedia.org)
  • Bubbling with carbon dioxide enriched air (1-2% CO2) can increase the growth rate, but may require additional buffer to maintain pH Selection is typically performed by antibiotic resistance genes. (wikipedia.org)
  • minimal
  • Within the −128/+46 segment, a novel element (called TβRE) was shown to be required for TGF-β1 inducibility of a reporter gene and, when multimerized, to be sufficient to transfer the inducibility to a minimal c- fos promoter ( 10 ). (jimmunol.org)
  • activate
  • Two signaling pathways arising from glucose metabolism converge to activate NFAT-mediated insulin gene transcription. (diabetesjournals.org)
  • In parallel to these observations in the insulin-producing INS-1 β-cell line, GLP-1 synergizes with glucose to activate expression of immediate-early response genes coding for transcription factors implicated in cell proliferation and differentiation, such as c- fos , c- jun , junB, zif-268, and nur-77 ( 5 , 19 ). (diabetesjournals.org)
  • Although all three CBFα family members activate the GL α promoter, only CBFα3 (AML-2) is induced by TGF-β1 in splenic B and I.29μ cells. (jimmunol.org)
  • These stimuli bind to heterotrimeric G protein-coupled receptors and rapidly activate immediate-early genes (c- fos , c- jun, egr-1 , etc.) that encode transcription factors via protein kinase C (PKC) pathway and mitogen-activated protein kinase (MAPK) phosphorylation cascade ( Bueno and Molkentin, 2002 ). (aspetjournals.org)
  • fragments
  • The organization and the structure of the genes encoding phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS) were determined and the effects of elicitor and suppressor produced by a pea pathogen, Mycosphaerella pinodes, were examined by transforming a chimeric DNA containing the PAL- and CHS-promoter fragments connected to a reporter gene into tobacco or pea plant with particle gun, Agrobacterium-mediated transformation, or electroporation. (nii.ac.jp)
  • CAT activity was also observed with fragments containing bases +65 to -193 and +65 to -117 in smooth-muscle cells. (biochemj.org)
  • In contrast with the smooth-muscle cells, no CAT activity was detected in Rat-1 fibroblasts with the smallest two fragments. (biochemj.org)
  • upstream
  • A region between −88 and −111 base pairs upstream from the UGT2B1 gene transcription start site contains a CCAAT enhancer binding protein (C/EBP)-like element and was previously shown by Dnase I footprint analysis to bind to proteins in both rat liver and human hepatoma (HepG2) cell nuclear extracts. (aspetjournals.org)
  • Class switching is effected by a deletional recombination that occurs between switch regions located upstream of each C H gene. (jimmunol.org)
  • On the basis of our findings, we conclude that downregulation of renin synthesis caused by interleukin-1 occurs primarily at the level of transcription and that DNA sequence or sequences mediating that effect are positioned within 4.1 kilobases upstream of the renin gene. (ahajournals.org)
  • We have cloned and sequenced a 1.9 kb fragment of the 5′-upstream sequence of the smooth-muscle-specific gene SM22 alpha. (biochemj.org)
  • The CYP3A4 gene exhibits a much more complicated upstream regulatory region in comparison with its paralogs. (wikipedia.org)
  • endogenous
  • These cells express high levels of renin mRNA from their endogenous renin gene and release approximately eightfold-more prorenin than active renin in culture. (ahajournals.org)
  • 6 7 8 The As4.1 cell line maintains high expression of its endogenous renin gene in addition to the transgene over long-term culture and demonstrates several features that are characteristic of kidney juxtaglomerular cells, including the ability to store and release active renin. (ahajournals.org)
  • 5 9 The endogenous renin gene expressed in the As4.1 cell line is Ren-1 c . (ahajournals.org)
  • This increased complexity renders the CYP3A4 gene more sensitive to endogenous and exogenous PXR and CAR ligands, instead of relying on gene variants for wider specificity. (wikipedia.org)
  • regulation
  • In this study, the importance of this region in the regulation of the UGT2B1 gene was assessed by functional and DNA binding assays. (aspetjournals.org)
  • This diverse expression pattern offers an opportunity to study the regulation of a single gene within different developmental contexts. (nih.gov)
  • This study reports for the first time a down-regulation of mdr1 gene transcription by a pharmacological agent. (aspetjournals.org)
  • Although the functions of GL transcripts are unknown, transcription of unrearranged C H genes is important for regulation of class switch recombination. (jimmunol.org)
  • Furthermore
  • Furthermore, two-point base pair mutations in any of the three identified NFAT sites within the rat insulin I promoter resulted in a significant reduction in the combined effect of glucose and GLP-1. (diabetesjournals.org)
  • Furthermore, the sequentially deleted promoter fragment of PSPAL1 connected to a plant expression vector containing the reporter gene (CAT) was introduced into pea protoplasts by electroporation, and induction by elicitor and suppression by orthovanadate was examined. (nii.ac.jp)
  • protein
  • We have previously identified NFAT (nuclear factor of activated T-cells) as a key regulator of insulin gene transcription in pancreatic β-cells that is activated by the calcium/calmodulin-dependent protein phosphatase 2B (calcineurin) in response to increased [Ca 2+ ] i ( 10 ). (diabetesjournals.org)
  • Reporter systems have been used for the determination of the efficiency of gene delivery systems, the intracellular localization of a gene product, the detection of protein-protein or protein-DNA interactions, the efficiency of translation initiation signals and the success of molecular cloning efforts. (wikipedia.org)
  • A common functional polymorphism in the promoter region of the microsomal triglyceride transfer protein gene influences plasma LDL levels. (ox.ac.uk)
  • Despite the importance of the TβRE for GL α transcription, no protein that binds the TβRE and regulates the GL α promoter has been identified. (jimmunol.org)
  • Protein quantification and the assessment of protein activity are also discussed, along with the centrifugation method and applications of PCR in forensics and paternity testing. (elsevier.com)
  • We show that a frameshift mutation near the 3' end prevents expression of the protein on the cell surface and reduces the rate of transcription of the corresponding gene. (cognizantcommunication.com)
  • The present invention further relates to process for cloning and expressing modified SAK gene fusion protein which imparts improved stability to the heterologous protein of interest. (freepatentsonline.com)
  • Novel self-cleavage activity of Staphylokinase fusion proteins: An interesting finding and its possible applications", Protein Expression and Purification, vol. 69, 2010, pp. 191-197. (freepatentsonline.com)
  • The ET-1-stimulated activator protein-1 (AP-1) DNA binding activity and AP-1-mediated reporter activity were attenuated by NO. In addition, NO also significantly inhibited ET-1-stimulated promoter activity of hypertrophic marker gene β-myosin heavy chain and the enhanced protein synthesis. (aspetjournals.org)
  • In humans, the CYP3A4 protein is encoded by the CYP3A4 gene. (wikipedia.org)
  • exon
  • Second, deletion of a DNA segment containing the promoter and first exon (I or GL exon) of the GL transcript by knockout technology abrogates switching to that C H gene ( 2 , 3 , 4 , 5 ). (jimmunol.org)
  • The region cloned consisted of the SM22 alpha promoter, a 65 bp exon containing most of the 5′-untranslated region and 307 bp of the first intron. (biochemj.org)
  • marker
  • It is important to use a reporter gene that is not natively expressed in the cell or organism under study, since the expression of the reporter is being used as a marker for successful uptake of the gene of interest. (wikipedia.org)
  • While this successfully removes the selective marker from the genome, it leaves FRT scars in place of the target gene. (wikipedia.org)
  • sequence
  • 1. A baculoviral vector for the production of an exogenous gene product in insect host cells wherein said host cells are incapable of permitting replication of an unmodified baculovirus which comprises an exogenous DNA sequence encoding the gene product wherein transcription of said DNA sequence is controlled by a promoter of an LTR selected from the group consisting of retroviral LTRs and transposable element LTRs. (patents.com)
  • A 1.5 kb fragment at the 5′ end of this sequence was able to drive the expression of a reporter chloramphenicol acetyltransferase (CAT) gene in both vascular smooth-muscle cells and Rat-1 fibroblasts. (biochemj.org)
  • In molecular biology, a reporter gene (often simply reporter) is a gene that researchers attach to a regulatory sequence of another gene of interest in bacteria, cell culture, animals or plants. (wikipedia.org)
  • induction
  • Since the regeneration line of pea p … More lant has not been established from the single cell lines, we obtained Km^R calli and the induction of GUS activities were examined by the treatment with fungal elicitor and the infection of pathogens. (nii.ac.jp)
  • Considerable induction of GUS activity was observed in some of the callus. (nii.ac.jp)
  • Within this segment resides a TGF-β1-responsive element (TβRE) that is required for induction of the promoter by TGF-β1 and, when multimerized, is sufficient to transfer TGF-β1 inducibility to another promoter. (jimmunol.org)
  • Mutation of both sites reduces but does not eliminate induction of the GL α promoter by TGF-β1 or by overexpression of CBF, possibly due to the presence of an additional CBF site in the promoter. (jimmunol.org)
  • Mutation of these motifs eliminates TGF-β1 induction of the GL α promoter. (jimmunol.org)
  • Together the data indicate that TGF-β1 induction of the α promoter involves induction of CBFα3, which binds to the TβRE of the promoter along with one or more proteins. (jimmunol.org)
  • It remains to be determined whether ET-1 induces NO production in cardiomyocytes and whether intracellular NO could modulate ET-1-induced cardiomyocyte hypertrophic response, including the induction of early response gene and re-expression of fetal gene program. (aspetjournals.org)
  • cluster
  • Thus, it has been proposed that Synechocystis originally possessed the ability to fix nitrogen gas, but lost the genes required for a fully functioning nitrogen fixation (nif) gene cluster. (wikipedia.org)
  • This gene is part of a cluster of cytochrome P450 genes on chromosome 7q21.1. (wikipedia.org)
  • cells
  • In this study, GLP-1 (100 nmol/l), in the presence of glucose (11 mmol/l), induced a ∼71-fold increase in insulin gene promoter activity in INS-1 pancreatic β-cells, an effect that was an order of magnitude larger than with either stimulant alone. (diabetesjournals.org)
  • These data suggest that the synergistic action of glucose and GLP-1 to promote insulin gene transcription is mediated through NFAT via PKA- and calcineurin-dependent pathways in pancreatic β-cells. (diabetesjournals.org)
  • The rate of insulin gene transcription in pancreatic β-cells is regulated by a complex integration of signals derived from nutrients, hormones, and neurotransmitters ( 1 - 5 ). (diabetesjournals.org)
  • To initiate such studies, we have cloned the gene encoding the human slow twitch skeletal muscle isoform of troponin I and have identified 5'-flanking sequences required for its expression in skeletal muscle cells. (nih.gov)
  • A 4-fold decrease in the mdr1 gene transcription rate was observed in the 30 microM VRP-treated CEM VLB100 cells. (aspetjournals.org)
  • Cytological studies showed that plants with Lux or GUS gene connected to 35S promoter as a control exhibited the distinctive GUS atibity in the cells possibly carrying the chimeric DNA, however those with PAL promoter did not exhibit GUS or Lux activities even by the treatment with fungal elicitor. (nii.ac.jp)
  • SRC-1 increased AhR-mediated, TCDD-dependent reporter gene activity threefold in Hepa-1 and COS-1 cells. (cognizantcommunication.com)
  • The residual promoter activity in the smallest fragment of the SM22 alpha promoter tested suggested that, unlike the smooth-muscle alpha-actin promoter, transcription from the SM22 alpha promoter can occur in smooth-muscle cells in the absence of factors binding to CC(A/Trich)6GG (CArG box) or CANNTG (E box) motifs. (biochemj.org)
  • Inhibitors of IF3 are expected to result in increased growth of specially-designed test cells due to higher levels of expression from the reporter gene. (google.es)
  • Cells cotransfected with the dominant negative and positive mutants of signaling molecules revealed that the Ras/Raf/extracellular-signal regulated kinase (ERK) signaling pathway is involved in ET-induced c- fos gene expression. (aspetjournals.org)
  • Only those cells that have successfully taken up the construct containing the CAT gene will survive and multiply under these conditions. (wikipedia.org)
  • CYP3A4 promotes the growth of various types of human cancer cell lines in culture by producing (±)-14,15-epoxyeicosatrienoic acids which stimulate these cells to grow, The cytochrome P450 is also reported to have fatty acid monooxgenase activity for metabolizing arachidonic acid to 20-Hydroxyeicosatetraenoic acid (20-HETE). (wikipedia.org)
  • deletion
  • Deletion analysis showed that a fragment of the promoter from +65 to -303 was more active in both cell types than the 1.5 kb fragment suggesting that there are silencer sequences in the region 5′ to the core promoter. (biochemj.org)
  • The λ-red recombineering system was published in 1998 and allows for insertion, deletion, or mutations to E. coli genes. (wikipedia.org)
  • exogenous
  • A novel viral expression vector is employed in the expression of exogenous genes in "non-permissive" hosts. (patents.com)
  • Exogenous genes under the control of a promoter which promotes expression in "non-permissive" hosts are thus used to expand the host range of the virus thus allowing production of polypeptides, including polypeptides which are toxic to the target host. (patents.com)
  • cell
  • Currently there is intense interest to define the mechanism of action of glucagon-like peptide-1 (GLP-1) in regulating β-cell function, including insulin gene transcription. (diabetesjournals.org)
  • We investigated the mechanism of verapamil (VRP) effects on mdr1 gene expression in two leukemic multidrug-resistant (MDR) cell lines, K562/ADR and CEM VLB100. (aspetjournals.org)
  • The promoter segment between −130 and +46, relative to the first initiation site for mouse GL α transcripts, is sufficient for expression and TGF-β1 inducibility of a reporter gene in B cell lines. (jimmunol.org)
  • Previous work showed that the promoter segment containing nucleotides from −128 to +46, relative to the first initiation site for GL α transcripts, is sufficient for expression and TGF-β1 inducibility of a reporter gene in two different B cell lines ( 10 ). (jimmunol.org)
  • We propose that expression of the antigen on the cell surface is part of a self-regulatory pathway that influences transcription of the corresponding gene. (cognizantcommunication.com)
  • The toxic effects are largely cell, tissue, and promoter context dependent. (cognizantcommunication.com)
  • By generating immunodeficient mice (in Rag-1(-/-) or nude background) deleted for PAI-1 gene (PAI-1(-/-)), we have evaluated the impact of host PAI-1 deficiency on the tumorigenicity of two malignant human skin keratinocyte cell lines HaCaT II-4 and HaCaT A5-RT3 forming low-grade and high-grade carcinomas, respectively. (ac.be)
  • b) detecting the translation of the reporter gene mRNA, wherein a specific increase in the translation of the reporter gene mRNA in the test cell contacted with the test compound relative to the translation of the reporter gene mRNA in a test cell not contacted with the test compound, indicates that the test compound inhibits IF3 in the test cell. (google.es)
  • Reporter genes are often used as an indication of whether a certain gene has been taken up by or expressed in the cell or organism population. (wikipedia.org)
  • To introduce a reporter gene into an organism, scientists place the reporter gene and the gene of interest in the same DNA construct to be inserted into the cell or organism. (wikipedia.org)
  • region
  • Ig class switching allows the recombined variable (VDJ) region gene of the heavy chain to be expressed with a new downstream constant region (C H ) 3 gene, resulting in a change in the effector function of the Ab. (jimmunol.org)
  • Previous studies have shown that mutually exclusive expression of Paramecium surface antigens can be overcome if two antigen genes contain the same 5' coding region. (cognizantcommunication.com)
  • In building the DNA construct, a segment of DNA coding for a flexible polypeptide linker region is usually included so that the reporter and the gene product will only minimally interfere with one another. (wikipedia.org)
  • proteins
  • Of particular interest among these proteins is Cyt1A, which synergizes and delays resistance to mosquitocidal Cry proteins and can be used to overcome resistance to Bs, as well as extend its spectrum of activity to, for example, the yellow fever mosquito Aedes aegypti ( Fig. 1 ). (biologists.org)
  • A more complex use of reporter genes on a large scale is in two-hybrid screening, which aims to identify proteins that natively interact with one another in vivo. (wikipedia.org)
  • enzyme
  • 3 Moreover, mice with both chromosomal copies of the IL-1β-converting enzyme gene inactivated were resistant to endotoxic shock. (ahajournals.org)
  • coli
  • Phage λ gamma is necessary to inhibit E. coli nuclease activity and protect the transformed linear DNA in vivo. (wikipedia.org)
  • stimulation
  • 20-HETE has a wide range of activities that also include growth stimulation in breast and other types of cancers (see 12-hydroxyeicosatetraenoic acid#cancer). (wikipedia.org)
  • tissues
  • The structure and the organization of three major CHS genes were determined from 38 CHS-cDNAs prepared from elicitor treated pea epicotyl tissues. (nii.ac.jp)
  • As a result, the reporter gene's expression is independent of the gene of interest's expression, which is an advantage when the gene of interest is only expressed under certain specific conditions or in tissues that are difficult to access. (wikipedia.org)
  • cytochrome
  • Resveratrol competitively inhibited, in a concentration-dependent manner, the activity of the carcinogen activating enzymes cytochrome P-450 (CYP)1A1/CYP1A2 in microsomes and intact HepG2 cells. (aspetjournals.org)
  • This gene is part of a cluster of cytochrome P450 genes on chromosome 7q21.1. (wikipedia.org)
  • CYP3A4 promotes the growth of various types of human cancer cell lines in culture by producing (±)-14,15-epoxyeicosatrienoic acids which stimulate these cells to grow, The cytochrome P450 is also reported to have fatty acid monooxgenase activity for metabolizing arachidonic acid to 20-Hydroxyeicosatetraenoic acid (20-HETE). (wikipedia.org)
  • organism
  • Among ≈4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. (pnas.org)
  • Reporter genes are often used as an indication of whether a certain gene has been taken up by or expressed in the cell or organism population. (wikipedia.org)
  • Many methods of transfection and transformation - two ways of expressing a foreign or modified gene in an organism - are effective in only a small percentage of a population subjected to the techniques. (wikipedia.org)
  • insertions
  • Transposon mutagenesis might overestimate the set by misclassification of nonessential genes that slow down the growth without arresting it but can also miss essential genes that tolerate transposon insertions ( 3 , 6 ). (pnas.org)
  • this technique is able to establish point mutations, gene deletions, and short sequence insertions in several genomic loci with increased efficiency and time sensitivity. (wikipedia.org)
  • bacteria
  • Most essential genes are present throughout a wide range of Bacteria, and almost 70% can also be found in Archaea and Eucarya. (pnas.org)
  • However, essential genes related to cell envelope, shape, division, and respiration tend to be lost from bacteria with small genomes. (pnas.org)
  • Sequence
  • You can either co-express two different genes of interest using systems that contain the pTRE3G-IRES Vector, which contains two multiple cloning sites flanking an IRES sequence (Cat. (clontech.com)
  • An upper-limit estimate of a minimal bacterial gene set was obtained from the sequence of the entire Mycoplasma genitalium genome, which contains only ≈480 genes ( 1 ). (pnas.org)
  • successfully
  • While this successfully removes the selective marker from the genome, it leaves FRT scars in place of the target gene. (wikipedia.org)
  • Only those cells that have successfully taken up the construct containing the CAT gene will survive and multiply under these conditions. (wikipedia.org)
  • systems
  • The EF-1 alpha promoter in these systems is derived from the human EEF1A1 gene that expresses the alpha subunit of eukaryotic elongation factor 1. (clontech.com)
  • growth
  • Analysis of the mutants, in conjunction with the literature data, leads us to conclude that there are only 271 genes indispensable for growth in LB when inactivated singly. (pnas.org)
  • 20-HETE has a wide range of activities that also include growth stimulation in breast and other types of cancers (see 12-hydroxyeicosatetraenoic acid#cancer). (wikipedia.org)
  • conditions
  • To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. (pnas.org)
  • Some glycolytic genes are regulated by the genesll1330 under light and glucose-supplemented conditions. (wikipedia.org)
  • Another
  • Another 79 genes were predicted to be essential. (pnas.org)
  • In building the DNA construct, a segment of DNA coding for a flexible polypeptide linker region is usually included so that the reporter and the gene product will only minimally interfere with one another. (wikipedia.org)
  • instead
  • however the Tet-On 3G transactivator is expressed from an EF1-alpha promoter instead of a CMV promoter. (clontech.com)
  • This increased complexity renders the CYP3A4 gene more sensitive to endogenous and exogenous PXR and CAR ligands, instead of relying on gene variants for wider specificity. (wikipedia.org)
  • responsible
  • Resveratrol exerts potent antioxidant and anti-inflammatory activities, which may be responsible for the beneficial effects of wine consumption in the prevention of cardiovascular disease ( Constant, 1997 ). (aspetjournals.org)
  • higher
  • More recently, an experimental approach involving high-density transposon mutagenesis of the H. influenzae genome led to a much higher estimate of ≈670 putative essential genes ( 3 ), whereas transposon mutagenesis of two mycoplasma species led to an estimate of 265-360 essential genes ( 4 ). (pnas.org)
  • approach
  • Functional proteomics approach to investigate the biological activities of cDNAs implicated in breast cancer. (dnasu.org)
  • A computational approach, based on the assumption that essential genes are conserved in the genomes of M. genitalium and Haemophilus influenzae , led to a description of a smaller set of some 260 genes ( 2 ). (pnas.org)
  • The approach used for gene inactivation has been described ( 8 ). (pnas.org)