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Expression analysis of an enrofloxacin and a tetracline resistant Escherichia coli strain

ABSTRACT: Comparison of the whole genome gene expression level of an enrofloxacin and tetracycline resistant E. coli strain with the wildtype it was derived from. The process of drug adaptation of E. coli MG1655 wildtype cells is further descibed in van der Horst, M, J.M. Schuurmans, M. C. Smid, B. B. Koenders, and B. H. ter Kuile (2011) in Microb. Drug Resist. 17:141-147. Resistance to amoxicillin was induced in E. coli by growth in the presence of stepwise increasing antibiotic concentrations. To investigate consequences of the aquisition of amoxicillin resistance the transcriptomic profile of sensitive and resistant cells was compared in the absence and presence of sub-inhibitory (0.25xMIC) amoxicillin concentrations was compared. Total RNA of 3 biological replicates of E. coli MG1655 wildtype cells and drug resistant cells cultured with (0.25xMIC) or without the drug was hybridized on a 12x135k custom designed microarraychip against one common reference.

ORGANISM(S): Escherichia coli str. K-12 substr. MG1655

SUBMITTER: Nadine HM-CM-$ndel

PROVIDER: E-GEOD-57084 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Expression analysis of an amoxicillin resistant Escherichia coli strain

Project description:Comparison of the whole genome gene expression level of an amoxicillin resistant E. coli strain with the wildtype it was derived from. The process of amoxicillin adaptation of E. coli MG1655 wildtype cells is further descibed in van der Horst, M, J.M. Schuurmans, M. C. Smid, B. B. Koenders, and B. H. ter Kuile (2011) in Microb. Drug Resist. 17:141-147. Resistance to amoxicillin was induced in E. coli by growth in the presence of stepwise increasing antibiotic concentrations. To investigate consequences of the aquisition of amoxicillin resistance the transcriptomic profile of sensitive and resistant cells was compared in the absence and presence of sub-inhibitory (0.25xMIC) amoxicillin concentrations was compared. Total RNA of 3 biological replicates of E. coli MG1655 wildtype cells and amoxicillin resistant cells cultured with (0.25xMIC) or without amoxicillin was hybridized on a 12x135k custom designed microarraychip against one common reference.

2013-05-27 | E-GEOD-47221 | biostudies-arrayexpress

Effect of multiple longitudinal biopsy sampling in individual mice

Project description:Small scale study to asses the effects of taking multiple skinbiopts from individual mice in time. Results showed no influence on expression levels in time from older samplings on newer samplings. skin biopts from 4 individual mice in taken at 6 points in time

2014-09-01 | E-GEOD-51347 | biostudies-arrayexpress

UV-B dose and time range finding study on mouse biopsy skin samples

Project description:In molecular biology, the design of mechanistic experiments has to be optimized by considering statistical and biological principles. In contrast to statistical principles, biological principles of experimental design are not universally formulated. In an attempt to pinpoint generally acceptable rules, we investigated the importance of determining the optimal ranges of scale of i.e. dose and time in gene expression experiments. We propose a protocol for executing small scale, genome wide, range finding studies, covering a wide range of the potentially relevant part of the design space to find the optimal ranges of experimentation. This protocol is executed and a proof-of-concept is presented, where this approach is tested for both an in-vitro and an in-vivo study that aim to unravel DNA repair mechanisms provoked after UV radiation. We identified four challenges of range finding studies in omics experimentation; (1) the modularity of biological processes, (2) their dynamics, (3) the extent to which end-points indicate biological processes, and (4) the costs associated with the assays, which are all addressed by our approach. 57 skin biopt samples taken from 12 individual mice on 8 timepionts and for 6 different UV-B doses. Per mouse 5 skin biopts were samples in time

2014-09-01 | E-GEOD-51348 | biostudies-arrayexpress

UV-C dose and time range finding study on Mouse Embryonic Fibroblasts (MEFs)

Project description:In molecular biology, the design of mechanistic experiments has to be optimized by considering statistical and biological principles. In contrast to statistical principles, biological principles of experimental design are not universally formulated. In an attempt to pinpoint generally acceptable rules, we investigated the importance of determining the optimal ranges of scale of i.e. dose and time in gene expression experiments. We propose a protocol for executing small scale, genome wide, range finding studies, covering a wide range of the potentially relevant part of the design space to find the optimal ranges of experimentation. This protocol is executed and a proof-of-concept is presented, where this approach is tested for both an in-vitro and an in-vivo study that aim to unravel DNA repair mechanisms provoked after UV radiation. We identified four challenges of range finding studies in omics experimentation; (1) the modularity of biological processes, (2) their dynamics, (3) the extent to which end-points indicate biological processes, and (4) the costs associated with the assays, which are all addressed by our approach. 48 MEF samples having various combinations of 9 timepoints and 6 UV-C Doses without replication were used

2014-09-01 | E-GEOD-50930 | biostudies-arrayexpress

The conserved DNA-binding protein WhiA is involved in cell division in Bacillus subtilis

Project description:One of the first steps in bacterial cell division is the polymerization of the tubulin-like protein FtsZ at midcell. The dynamics of FtsZ polymerization is regulated by a set of proteins among which ZapA. A zapA mutation does not result in a clear phenotype in Bacillus subtilis. In this study we used a synthetic-lethal screen to find genes that become essential when ZapA is absent. Three transposon insertions were found in yvcL. Deletion of yvcL in a wild type background had only a mild effect on growth, but a yvcL zapA double mutant is very filamentous and sick. This filamentation is caused by a strong reduction in FtsZ polymerization, suggesting that YvcL is involved in an early stage of cell division. YvcL is 25 % identical and 50 % similar to the Streptomyces coelicolor transcription factor WhiA. WhiA is required for septation of aerial hyphae during sporulation. Using GFP fusions, we show that YvcL localizes at the nucleoid. Surprisingly, transcriptome analyses in combination with a ChIP on chip assay did not provide clear evidence that YvcL functions as a transcription factor. To gain more insight into the function of YvcL, we searched for suppressors of the filamentous phenotype of a M-bM-^HM-^FyvcL M-bM-^HM-^FzapA mutant. Transposon insertions in gtaB and pgcA restored normal cell division of the double mutant. The corresponding proteins have been implemented in the metabolic sensing of cell division. We conclude that YvcL (WhiA) is involved in cell division in B. subtilis through an as yet unknown mechanism. Comparing wild tpe Bascillus subtilus (n=3) with Bascillus Subtilis KS400 (n=2) and Bascillus subtilis KS696 (n=2)

2013-11-28 | E-GEOD-45824 | biostudies-arrayexpress

Molecular signatures of the evolving immune response in mice following a Bordetella pertussis infection.

Project description:Murine gene expression responses to Bordetella pertussis were determined in lung and spleen, between 0 and 28 days post infection. Data were compared to other parameters such as microarray, flow cytometry, multiplex immunoassays, and lung clearance. A 57 array study with 27 arrays for lung and 30 for spleen. For each tissue we used a common reference made from RNA samples of the same tissue. Please note that the reference samples are made by pooling RNA of all samples used for a tissue (either infected or uninfected). Thus, Lung and spleen each have their own reference.

2014-09-02 | E-GEOD-53294 | biostudies-arrayexpress

Analysis of SoxNeuro gene expression changes, genome-wide binding, and comparison of SoxNeuro and Dichaete binding profiles in wild type and mutant embryos

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2014-04-23 | E-GEOD-47338 | biostudies-arrayexpress

Binding in wild type, D and SoxN mutant embryos: SoxN vs D.

Project description:Genome-wide binding of transcription factors SoxN and D in stage 12-17 wild type embryos (SoxNDam and DDam), of SoxN in D mutants (D-SoxNDam) and of D in SoxN mutants (SoxN-DDam). Four DamID experiments. Three biological replicates per experiment. Two conditions: Dam (control) vs SoxNDam or DDam or D-SoxNDam or SoxN-DDam (sample)

2014-04-23 | E-GEOD-47228 | biostudies-arrayexpress

Expression analysis of an amoxicillin resistant Escherichia coli strain

2013-05-27 | GSE47221 | GEO

Binding in wild type embryos: SoxN.

Project description:Genome-wide binding of SoxN in wild type embryos: SoxNDam (DamID, stage 8-11), SoxND1 (ChIP, stage 8-11), SoxND2 (ChIP, stage 8-11), SoxNPA179 Early (ChIP, stage 7-10), and SoxNPA179 Late (ChIP, stage 11-13). One DamID experiment, four ChIP experiments. Three biological replicates per experiment. Two conditions: Dam (control) vs SoxNDam (sample) for DamID, anti-M-NM-2gal (control, 40-1a antibody from DSHB) vs anti-SoxN (sample, either SoxND1, SoxND2 or SoxNPA179 antibody) for ChIP. The only commercially available antibody is the anti-BetaGal. All the anti-SoxN antibodies are home-made. anti-BetaGal: Developmental Studies Hybridoma Bank (DSHB), 40-1a, mouse, monoclonal (http://dshb.biology.uiowa.edu/beta-galactosidase_2) SoxND1: polyclonal antiserum raised in rabbit immunized with a protein fragment corresponding to amino acids 2-92 of SoxN. Produced by the modENCODE consortium (http://www.modencode.org/). SoxND2: polyclonal antiserum raised in rabbit immunized with a protein fragment corresponding to amino acids 317-417 of SoxN. Produced by the modENCODE consortium (http://www.modencode.org/). SoxNPA179: affinity purified polyclonal antibody raised in rabbit immunized with a small peptide corresponding to amino acids 506-521 of SoxN. Commisioned by our lab and produced by Eurogentec (http://www.eurogentec.com/eu-home.html).

2014-04-23 | E-GEOD-47337 | biostudies-arrayexpress

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