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Global gene expression in Moraxella catarrhalis ATCC 43617 grown under iron-replete and iron-limiting conditions

ABSTRACT: Total RNA samples were isolated from M. catarrhalis grown in BHI (iron-replete condition, designated DF0) and in BHI containing 30 mM Desferal (iron-limiting condition, designated DF30). Total RNA and M. catarrhalis genome-directed primers (GDPs) were used for synthesis of Cy3- or Cy5-labeled cDNA samples. Three independent M. catarrhalis growth experiments and total RNA isolations were used for synthesis of cDNA samples, and a total of six DNA microarray hybridizations were performed in this study. Keywords: Effect of Growth Environment on Global Gene Expression by Moraxella catarrhalis

ORGANISM(S): Moraxella catarrhalis

PROVIDER: GSE4346 | GEO | 2007/12/07

SECONDARY ACCESSION(S): PRJNA94841

REPOSITORIES: GEO

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Identification of genes essential for Moraxella catarrhalis growth under iron-limiting conditions using Genomic Array Footprinting (GAF): Expression analysis of directed gene deletion mutants.

2013-06-06 | GSE41547 | GEO

Identification of genes essential for Moraxella catarrhalis growth under iron-limiting conditions using Genomic Array Footprinting (GAF): Expression analysis of directed gene deletion mutants.

Project description:Iron-sequestration by the human host is a first line defense against respiratory pathogens like Moraxella catarrhalis, which consequently experiences a period of iron-starvation during colonization and infection. We determined the genetic requirements for M. catarrhalis growth during iron-starvation using the high-throughput genome-wide screening technology genomic array footprinting (GAF). To further characterize the functional consequences of the gene deletions (GAF identified genes), we determined transcriptome profiles of the wild-type and directed mutant strains during exponential growth in BHI medium. The results described in this study are further discussed in Stefan P.W. de Vries, Peter Burghout, Jeroen D. Langereis, Aldert Zomer, Peter W.M. Hermans, Hester J. Bootsma: Genetic requirements for Moraxella catarrhalis growth under iron-limiting conditions, Molecular Microbiology. M. catarrhalis BBH18 directed mutants (n = 3) and their parental wild-type BBH18 strain (n = 4) were expanded to mid-log phase (OD620nm, 1.2 to 1.4) and RNA was isolated as previously described (de Vries et al., 2010 J Bacteriol 192: 3574-3583). Total RNA was labeled according to standard Nimblegen gene expression array protocols and hybridized to 4x72K custom design Nimblegen arrays for read-out.

2013-06-06 | E-GEOD-41547 | biostudies-arrayexpress

Identification of genes essential for Moraxella catarrhalis growth under iron-limiting conditions using Genomic Array Footprinting (GAF)

Project description:Iron-sequestration by the human host is a first line defense against respiratory pathogens like Moraxella catarrhalis, which consequently experiences a period of iron-starvation during colonization and infection. We determined the genetic requirements for M. catarrhalis growth during iron-starvation using the high-throughput genome-wide screening technology genomic array footprinting (GAF). To this end, a large marinerT7 transposon mutant library (~28,000 independent transposon mutants) was grown under iron-limiting conditions, achieved by sequestration of iron by 30 µM Desferal (DF30), and under control growth conditions (brain heart infusion broth, DF0). Mutants were recovered at exponential- and the early-stationary growth phase and used for the generation of mutant-specific cDNA probes that were hybridized to custom-designed NimbleGen GAF microarrays. The results described in this study are further discussed in Stefan P.W. de Vries, Peter Burghout, Jeroen D. Langereis, Aldert Zomer, Peter W.M. Hermans, Hester J. Bootsma: Genetic requirements for Moraxella catarrhalis growth under iron-limiting conditions, Molecular Microbiology.

2013-06-06 | GSE41546 | GEO

Identification of genes essential for Moraxella catarrhalis growth under iron-limiting conditions using Genomic Array Footprinting (GAF)

Project description:Iron-sequestration by the human host is a first line defense against respiratory pathogens like Moraxella catarrhalis, which consequently experiences a period of iron-starvation during colonization and infection. We determined the genetic requirements for M. catarrhalis growth during iron-starvation using the high-throughput genome-wide screening technology genomic array footprinting (GAF). To this end, a large marinerT7 transposon mutant library (~28,000 independent transposon mutants) was grown under iron-limiting conditions, achieved by sequestration of iron by 30 M-BM-5M Desferal (DF30), and under control growth conditions (brain heart infusion broth, DF0). Mutants were recovered at exponential- and the early-stationary growth phase and used for the generation of mutant-specific cDNA probes that were hybridized to custom-designed NimbleGen GAF microarrays. The results described in this study are further discussed in Stefan P.W. de Vries, Peter Burghout, Jeroen D. Langereis, Aldert Zomer, Peter W.M. Hermans, Hester J. Bootsma: Genetic requirements for Moraxella catarrhalis growth under iron-limiting conditions, Molecular Microbiology. cDNA probes generated from mutants recovered during the exponential growth phase, or early-stationary phase under challenge (iron-limiting, DF30) or control conditions (DF0). Hybridization on GAF 4x72K custom design Nimblegen GAF arrays for read-out.

2013-06-06 | E-GEOD-41546 | biostudies-arrayexpress

Global gene expression in Moraxella catarrhalis ATCC43617 cells grown in the planktonic and in biofilm states

Project description:Total RNA samples were isolated from M. catarrhalis grown in the planktonic state (i.e., in broth) and in a biofilm. Biofilms were generated by growing this strain in the continuous-flow Sorbarod filter system for three days. Total RNA samples and M. catarrhalis genome-directed primers (GDPs) were used for synthesis of Cy3- or Cy5-labeled cDNA samples. Five independent M. catarrhalis growth experiments and total RNA isolations were used for the synthesis of cDNA samples, and a total of six DNA microarray hybridizations were performed in this study. Keywords: Effect of Growth Environment on Global Gene Expression by Moraxella catarrhalis

2007-12-07 | GSE4348 | GEO

SWATH-MS proteome of Moraxella catarrhalis under iron restricted conditions

Project description:Moraxella catarrhalis is a human-restricted bacterial pathogen that causes otitis media in children and exacerbations of chronic obstructive pulmonary disease in adults. Iron is a co-factor required for the function of proteins involved in respiration and DNA replication, and thus is essential to life for most bacterial species. As iron is sequestered by host tissues, all host-adapted pathogens encounter periods of iron starvation. Previous studies have defined the repertoire of genes required for growth of M. catarrhalis under iron restricted conditions, and described differential expression at the transcriptional level. However, global changes in protein expression in response to iron starvation have not been investigated. Here we provide a SWATH-MS dataset describing differential expression of the M. catarrhalis CCRI-195ME proteome under iron restricted versus iron replete conditions.

2020-01-24 | PXD017206 | Pride

Gene Expression Profile of Moraxella catarrhalis Grown in Pooled Human Sputum

Project description:Comparison of the gene expression profile of Moraxella catarrhalis grown in the presence of 20% pooled human sputum in chemically-defined medium relative to Moraxella catarrhalis grown in chemically-defined medium alone. Moraxella catarrhalis ATCC43617 was grown to mid-logarithmic phase either in the presence of 20% pooled human sputum in chemically-defined medium or in chemically-defined medium alone. Total RNA was extracted from bacterial cells exposed to each of these conditions and cDNA was generated for CyDye labelling. 3 biologic replicates were generated and each replicate underwent a dye swap (total of 6 experimental data collections). The gene expression profile reported is that of Moraxella catarrhalis grown in the presence of pooled human sputum in a chemically-defined medium relative to Moraxella catarrhalis grown only in the presence of the chemically-defined medium.

2014-02-02 | E-GEOD-27059 | biostudies-arrayexpress

Expression profiling of Moraxella catarrhalis RH4 during in vitro growth in brain heart infusion (BHI) medium

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2010-05-07 | E-GEOD-21632 | biostudies-arrayexpress

M. catarrhalis Gene Expression in the Chinchilla Nasopharynx

Project description:M. catarrhalis strain O35E.rpsl was inoculated into the nasopharynx of healthy, male adult chinchillas. 24 hours later the nasopharyngeal tissues were extracted and homogenized. Total RNA was extracted from these tissue samples. Subsequently, M. catarrhalis genome directed primers were utilized to synthesize cDNA from the total RNA sample. As a control, M.catarrhalis strain O35E.rpsl was grown in BHI broth to a Klett density of 200 units and underwent RNA extraction per standard protocols. The genome directed primers mentioned above were utilized to synthesize cDNA. Both cDNA samples were subsequently labelled with either Cy3 or Cy5 and hybridized to a custom Microarrays, Inc. gene chip and scanned after 16 hours. Differential gene expression was measured utilizing the broth grown cells as the baseline and the chinchilla isolated cells as the experimental variable.

2012-04-04 | GSE13559 | GEO

Gene Expression Profile of Moraxella catarrhalis Grown in Pooled Human Sputum

2014-02-02 | GSE27059 | GEO

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