Transcriptomics,Genomics

Dataset Information

27

Transcriptome Changes in Mycoplasma hyopneumoniae During Infection


ABSTRACT: Mycoplasma hyopneumoniae causes swine pneumonia and contributes significantly to porcine respiratory disease complex. The mechanisms of pathogenesis are difficult to address since there is a lack of genetic tools, but microarrays can be used to study transcriptional changes that occur during colonization and disease in pigs. This approach has the potential to identify genes important to virulence. This study sought to identify genes that change transcript levels during infection. To accomplish this, organisms collected from bronchial alveolar lavages were compared to that of broth grown organisms. Bronchial alveolar lavage was performed on pigs 28 days post infection with M. hyopneumoniae, and organisms were isolated by differential centrifugation. Mycoplasma RNA enriched preparations were then obtained from total RNA by subtracting eucaryotic ribosomal and messenger RNAs. cDNA was generated with mycoplasma ORF-specific primers, fluorescently labeled with Cy3 and Cy5, and used to interrogate microarrays. Arrays were scanned and analyzed using a mixed linear statistical model. Nine biological replicates were analyzed in this fashion. Our analysis indicated that 33 Mhyo genes were up-regulated and 46 genes were down-regulated (p<0.01) during disease in the pig lung at a false discovery rate < 2.7%. Of the down-regulated genes, 27 of 46 (59%) lacked assigned function, and 20 of 33 (61%) of the up-regulated genes were hypothetical genes. Four down-regulated and two up-regulated genes were putative lipoproteins. secA (mhp295; p = 0.003), and two glycerol transport permeases (potA (mhp380); p = 0.006 and ugpA (mhp381); p = 0.003) were up-regulated in vivo. Elongation factor EF-G (fusA (mhp083); p = 0.002), rpoC (mhp635; p = 0.003), adenylate kinase (adk (mhp208); p = 0.001), prolyl aminoacyl tRNA synthetase (proS (mhp397); p = 0.009) and cysteinyl-tRNA synthetase (cysS (mhp661); p < 0.001) were down-regulated in vivo. Keywords: RNA, spotted DNA/cDNA Overall design: Nine independent BALF samples were processed for total RNA along with nine in vitro cultures. Each in vivo BALF sample was paired with an independent in vitro culture sample for hybridization on nine two color arrays. For five of the arrays, the control in vitro sample was labeled with Alexa Fluor 555 and compared to Alexa Fluor 647-labeled BALF sample. The dye assignment to control samples was reversed for the other 4 arrays. The slides were hybridized under identical conditions as described under hyb protocol.

INSTRUMENT(S): Mycoplasma hyopneumoniae spotted PCR array

SUBMITTER: Chris Minion  

PROVIDER: GSE9057 | GEO | 2008-01-01

SECONDARY ACCESSION(S): PRJNA102571

REPOSITORIES: GEO

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Transcriptome changes in Mycoplasma hyopneumoniae during infection.

Madsen Melissa L ML   Puttamreddy Supraja S   Thacker Eileen L EL   Carruthers Michael D MD   Minion F Chris FC  

Infection and immunity 20071210 2


Mycoplasma hyopneumoniae causes swine pneumonia and contributes significantly to the porcine respiratory disease complex. The mechanisms of pathogenesis are difficult to address, since there is a lack of genetic tools, but microarrays are available and can be used to study transcriptional changes that occur during disease as a way to identify important virulence-related genes. Mycoplasmas were collected from bronchial alveolar lavage samples and compared to broth-grown cells using microarrays. B  ...[more]

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