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ERX034839: 454 GS Junior sequencing
1 LS454 (454 GS Junior) run: 123,943 spots, 111.1M bases, 255.9Mb downloads

Design: Formula fed 3 month old baby sample
Submitted by: Texas A&M University
Study: Mutualism between gut microbiota and the host as revealed in a comparative study of breast-fed versus formula-fed infants.
show Abstracthide Abstract
Ongoing efforts are directed at understanding the mutualism between the gut microbiota and the host in breast-fed versus formula-fed infants. Due to the lack of tissue biopsies, no investigators have performed a global transcriptional analysis of the developing intestine in healthy infants. As a result, the crosstalk between the microbiome and the host transcriptome in the developing mucosal-commensal environment has not been determined. In this study, we examined the host intestinal mRNA gene expression and microbial DNA profiles in full term 3 month-old infants exclusively formula fed (FF; n=6) or breast fed (BF; n=6) from birth to 3 months. Host mRNA microarray measurements were performed on intact sloughed epithelial cells in stool samples collected at 3 months. Microbial composition from the same stool samples was assessed by metagenomic pyrosequencing. Both host mRNA expression and bacterial microbiome phylogenetic profiles provided strong feature sets that classified the two groups of babies (FF and BF). To determine the relationship between epithelial cell gene expression and the bacterial colony profiles, the host transcriptome and functionally profiled microbiome data were analyzed in a multivariate manner. From a functional perspective, analysis of the gut microbiota's metagenome revealed that virulence characteristics differed between the FF and BF babies. Using canonical correlation analysis, evidence of multivariate structure relating eleven host immunity/mucosal defense-related genes and microbiome virulence characteristics was observed. These results, for the first time, provide insight into the integrated responses of the host and microbiome to dietary substrates in the early neonatal period.
Sample: Stool sample from a 3 month old baby whose diet was only formula from birth. Genomic DNA was extracted using a modification of the method of Yu and Morrison. Deviation from the protocol included the use of glass beater steps. Briefly, 250 mg (wet weight) of feces was weighed into a 2 mL tube containing glass matrix E (MP Biomedicals, Solon, OH). One mL of lysis buffer (500 mM NaCl, 50 mM Tris-HCl, 50 mM EDTA, 4% sodium dodecyl sulfate) was added to the tube and shaken for 30 sec. Samples were then incubated at 70°C for 15 min. After centrifugation at 16,000 g for 5 min, supernatants were collected into 2.0 mL tubes. Lysis buffer (300 µL) was subsequently added and the above steps were repeated. Nucleic acids in the supernatant were precipitated sequentially with ammonium acetate and isopropanol, and dissolved in TE buffer. The precipitated nucleic acids were then treated with DNase-free RNase, proteinase K, and further purified on a QIAamp spin column from QIAgen DNA Mini Stool Kit (Qiagen, Valencia, CA). DNA
SAMEA1317683 • ERS075410 • All experiments • All runs
Library:
Name: ZZ03-007
Instrument: 454 GS Junior
Strategy: WGS
Source: METAGENOMIC
Selection: RANDOM
Layout: SINGLE
Spot descriptor:
          
Group tagBasecalls
0ACGTACACACT
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Runs: 1 run, 123,943 spots, 111.1M bases, 255.9Mb
Run# of Spots# of BasesSizePublished
ERR056995123,943111.1M255.9Mb2012-04-18

ID:
173361

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