Comparison of IS5 Deletion mutant to E.coli Motile and NonMotile Isolates
Ontology highlight
ABSTRACT:
ORGANISM(S): Escherichia coli
SUBMITTER: Betsy Martinez-Vaz
PROVIDER: E-GEOD-2694 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:human bronchial smooth muscle cells were infected with adenoviral constructs containing either the phosphorylation mutant form of HSP27 (HSP27-3A) or the wild type form (HSP27-WT) at 40 MOI for 4 days. All sets were stimulated with a cytokine mixture containing 10 ng/ml IL-1beta, TNF-alfa, and gamma-IFN for 20 hours prior to harvest. This series represents 4 separate preparations of cells and RNA, with each set being subjected to a dye flip and hybridized to 2 different slides.
2010-06-09 | E-GEOD-1292 | biostudies-arrayexpress
Project description:human bronchial smooth muscle cells were growth arrested. One group was stimulated with 10ng/mL of IL-1beta, TNF-alfa, and gamma IFN for 20 hours. For every cDNA sample, 2 dyeflipped slides were hybridized (these have the same date in the titles). All cy3 (channel 2) measurements were normalized to the dye coupling control before a ratio of the values was taken.
2010-06-09 | E-GEOD-1288 | biostudies-arrayexpress
Project description:Antipsychotic drugs are classified as typical and atypical based on extrapyramidal effects. However, since the frontal cortex is one of the most important regions for antipsychotic actions, this study attempted to classify antipsychotic drugs based on gene expression in the frontal cortex. Chlorpromazine and thioridazine were selected as typical antipsychotics, and olanzapine and quetiapine as atypical antipsychotics. Since these drugs have similar chemical structures, the effect of the basic structure on gene expression can be eliminated. Cluster analysis of microarray experiments showed thioridazine and olanzapine constituted a robust cluster. K-means clustering separated 4-drug-administered mice into chlorpromazine-quetiapine and thioridazine-olanzapine groups. This classification scheme is different from that which is based on criteria currently used to group the typical and atypical drugs and suggests that antipsychotic drugs can be further separated into multiple groups. Experiment Overall Design: Male 13-week-old ddY mice (Japan SLC Co., Hamamatsu, Japan) weighing 38-43 g were used. Animals were housed with free access to standard food in an air-conditioned room under a constant dark-and-light cycle (light: 7:00 a.m. to 7:00 p.m.) at a temperature of 22 to 24°C and 60 to 70% relative humidity. Ether was used for anesthesia during decapitation. All efforts were made to minimize animal suffering and to reduce the number of animals used. The present experiments were carried out after obtaining permission from the Committee of Animal Experimentation of the Graduate School of Medical and Dental Sciences at Kagoshima University. Experiment Overall Design: Doses of 25 mg/kg chlorpromazine HCl (Sigma-Aldrich Co., St. Louis, MO), 25 mg/kg thioridazine HCl (Sigma-Aldrich Co.), 1.25 mg/kg olanzapine (gift from Eli Lilly and Company, Indianapolis, IN) and 18.75 mg/kg quetiapine fumarate (gift from AstraZeneca, Macclesfield, UK) were used in the study. The dosages for these drugs were determined from therapeutically equivalent doses previously reported (Lehman et al, 2003; Woods, 2003). The drugs were dissolved in acetic anhydride and diluted with 0.9% saline, resulting in a final concentration of acetic acid of 0.5%. The drugs were injected intraperitoneally once daily for 28 consecutive days in a volume of 0.1 ml/10 g body weight. Experiment Overall Design: Microarray experiments were performed using an Agilent G4121A Mouse Oligo Microarray Kit (Agilent Technologies, Palo Alto, CA) as per the manufacturerâ??s instructions. The frontal cortex was immediately removed and the RNA was stabilized in RNAlater RNA Stabilization Reagent (Qiagen, Valencia, CA) and stored at -80°C until use. Total RNA was extracted using the RNeasy Mini Kit (Qiagen). The RNA was amplified and labeled by the Low RNA Input Fluorescent Linear Amplification Kit (Agilent). To synthesize cDNA, 200 ng total RNA was used. Vehicle-injected controls were labeled by cyanine 3 (PerkinElmer Life Sciences, Inc., Boston, MA) and drug-injected mice were labeled by cyanine 5 (PerkinElmer Life Sciences, Inc.). Hybridizations to the microarray were performed using the In situ Hybridization Kit Plus (Agilent). Doses of 750 ng cyanine 3-labeled cRNA, 750 ng cyanine 5-labeled cRNA, and control targets were mixed and fragmented in the kitâ??s fragmentation buffer, and then hybridized to the microarrays for 17 hours at 60°C in a hybridization rotator (Agilent) set to rotate at 4 rpm. Microarrays were washed in 6Ã?SSC with 0.005% Triton X-102 at RT for 10 min, followed by 0.1Ã?SSC with 0.005% Triton X-102 at 4°C for 5 min. The slides were dried, and then scanned by the Agilent G2565BA Microarray Scanner System. Data were analyzed using the Agilent Feature Extraction Software version 7.1. A rank consistency filter and LOWESS were used for dye normalization. Control mice and drug-injected mice were processed in parallel. The data discussed in this publication are presented in accordance with the MIAME guidelines (http://www.mged.org/Workgroups/MIAME/miame.html) and were deposited in NCBIs Gene Expression Omnibus (GEO, http://www.ncbi.nlm.nih.gov/geo/). They can be accessed using the GEO series accession number GSE1501. Experiment Overall Design: Cluster analysis was performed using free software written by M. Eisen (http://rana.lbl.gov/EisenSoftware.htm). The cclust package in the â??Râ?? statistical software system (www.cran.r-project.org) was used to find the most appropriate number of clusters (i.e. â??kâ?? in the k-means clustering).
2008-06-12 | E-GEOD-1501 | biostudies-arrayexpress
Project description:Right ventricular free wall (RVFW) pacing results in left ventricular dyssynchrony with early septal shortening followed by late lateral contraction that reciprocally stretches the septum. Dyssynchrony is disadvantageous to cardiac mechano-energetics, yet little is known about its molecular consequences. We tested the hypothesis that dyssynchrony selectively alters regional gene expression in mice, employing a novel miniature implantable cardiac pacemaker. Mice were subjected to 1-week overdrive RVFW pacing (720 min-1, baseline HR 520-620 min-1) to induce dyssynchrony (pacemaker: 3V lithium battery, rate programmable, 0.8 grams, bipolar lead). Electrical capture was confirmed by pulsed-wave Doppler at implantation and terminal study, and dyssynchrony by echocardiography. Gene expression from left ventricular septal and lateral-wall myocardium were assessed by microarray (dual-dye method, Agilent) using oligonucleotide probes and dye swap. Identical analysis was applied to 4 synchronously contracting controls. Of 22,000 genes surveyed, only 18 genes displayed significant (p<0.01) differential expression between septal/lateral walls exceeding 1.5-fold relative to any disparities in synchronous controls. These changes were confirmed by qPCR with excellent correlations. Most (16) of the genes showed greater septal expression. Of particular interest were 7 genes coding proteins involved with stretch responses, matrix remodeling, stem cell differentiation to myocyte lineage, and Purkinje fiber differentiation. One-week cardiac dyssynchrony triggers regional differential expression differences in relatively few select genes. Such analysis using a murine implantable pacemaker should facilitate molecular studies of cardiac dyssynchrony and help elucidate novel mechanisms by which stress/stretch stimuli due to dyssynchrony impact the normal and failing heart. Experiment Overall Design: Left ventricle segments from the mouse heart - septal and lateral, were isolated from 4 dyssynchronous mice that were kept separate. RNA from the synchronous mice were pooled into a control septal and lateral sample. Functional genomic analysis was conducted on these 10 RNA samples with fluorophore reversal, such that each sample was assayed on two different microarrays. Corresponding septal and lateral samples from the same heart were paired on the same microarray to measure the relative differences in gene expression between the different regions of the heart then differences were compared across multiple mice to find overlapping genes that were affected by the pacememaker implantation.
2008-06-12 | E-GEOD-3535 | biostudies-arrayexpress
Project description:PAMP (Pathogen-Associated Molecular Pattern) recognition plays an important role in innate immune responses both in plants and animals. Lipopolysaccharides (LPS) of gram-negative bacteria are a typical PAMP molecule and have been reported to induce defense-related responses such as suppression of hypersensitive responses, defense gene espression and systemic resistance in plant. However the detailed analysis of these cellular responses and the molecular machinery involved in the perception and transduction of LPS molecule largely remains to be studied. Furthermore, the biological activities of LPS on plants have so far been reported only for dicots and no information is available on the action of LPS on monocots. We report here that bacterial LPSs, both from plant pathogens and non-pathogens, could induce various defense responses in rice cells, including reactive oxygen generation and defense gene expression. Global analysis of gene expression induced by two PAMP elicitors, LPS and chitin oligosaccharide elicitor, showed a close correlation between the gene responses induced by these elicitors, indicating the convergence of signaling cascades downstream of corresponding receptors. Further, we show that the defense responses induced by LPS are associated with programmed cell death, a finding so far not reported for LPS action on plant cells. Experiment Overall Design: 1. LPS treatment (WT), 2. LPS treatment (WT) color-swap, 3. N-acetylchitooctaose treatment (WT), 4. N-acetylchitooctaose treatment (WT) color-swap
2008-11-06 | E-GEOD-5906 | biostudies-arrayexpress
Project description:human bronchial smooth muscle cells were either infected with empty adenoviral type 5 vector at 40 MOI for 4 days, or Mock infected. Each sample set was then stimulated with a mixture of 10ng/mL IL-1beta, TNF-alfa, and gamma-IFN for 20 hours prior to RNA harvest. RNA samples were split, and the series consists of 3 slides, 2 of which are dye flipped.
2010-06-09 | E-GEOD-1291 | biostudies-arrayexpress
Project description:Fhl1-9myc ChIP-chip, YPD, OD600=0.8, 2 arrays with duplicate spotting of yeast intergenic regions. AND Ifh1-9myc ChIP-chip, cells grown in YPD, OD600=0.8, 2 arrays with duplicate spotting of yeast intergenic regions. Keywords = Fhl1
2010-06-05 | E-GEOD-1930 | biostudies-arrayexpress
Project description:The paralogous transcription factors Aft1p and Aft2p activate the expression of genes involved in iron metabolism under iron depleted conditions. Both are able to bind to the same DNA consensus sequence in vitro. We used DNA microarrays and loss of function mutant strains to better understand the respective roles of Aft1p and Aft2p in the regulation of gene expression
2010-06-09 | E-GEOD-1763 | biostudies-arrayexpress
Project description:The parent S. cerevisiae strain, BY4741, and specific gene deletions of the parent S.cerevisiae strain BY4741 were treated with 100uM NaAsO2 (AsIII) for 2h and compared to the untreated counterpart (yap1Î? vs. yap1Î? 2h 100uM AsIII, cad1Î? vs. cad1Î? 2h 100uMAsIII, rpn4Î? vs. rpn4Î? 2h 100uM AsIII, arr1Î? vs. arr1Î? 2h 100uM AsIII, parent vs. parent with 2h 100uM AsIII). The untreated specific deletion strain was also compared to the untreated parent strain in order to identify differential expression of genes due to the knockout alone. Keywords: other
2010-06-05 | E-GEOD-1798 | biostudies-arrayexpress
Project description:We report here an experimental search for small non-coding RNAs in a low GC, gram-positive, human pathogenic bacteria Streptococcus pneumoniae. Based on the bioinformatic analyses by Livny et al. (2006, Nucleic Acids Res. 34:3484-93), we tested 36 candidates by Northern analyses and confirmed the expression of 8 novel and one previously-reported sRNAs (CcnA) in the genome of D39 serotype 2 strain. Some of the sRNAs showed differential expression in stationary phase or after treatment with competence stimulatory peptide compared to untreated exponential cells. One sRNA we detected is CcnA, one of five CiaR-controlled non-coding RNA (CcnA to E) reported previously. By ectopically expressing this sRNA in a ÎciaR strain, we provide evidence that CcnA plays important roles in the regulation of growth and chain length of S. pneumoniae. Microarray analysis comparing global gene expression of CcnA-expressing and CcnA-nonexpressing ÎciaR strains showed significant differences in expression of competence related genes and five putative transcriptional regulators. QPCR and transformation assays further confirmed that CcnA sRNA mediates a strong negative effect on competence development. Our findings suggested many of the CiaRH function may be mediated via Ccn sRNAs and that sRNAs play regulatory roles in S. pneumoniae, an organism that lacks Hfq RNA chaperone. Three independent hybridizations using independent RNA preparations from the strain IU2678 (a ciaR knockout strain with no ccnA expression, reference strain) and strain IU2841 (a ciaR knowckout strain with ccnA expression at an ectopic site) were performed. Dye swap was performed with the reference strain labeled with Cy3 in two hybridizations and Cy5 in the one hybridizations.
2010-05-14 | E-GEOD-14688 | biostudies-arrayexpress