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EDL933 cultured in bovine small intestine content (BSIC) vs M9 minimal media

ABSTRACT: Enterohemorrhagic Escherichia coli (EHEC) are transmitted from cattle to human by means of contaminated food products resulting from fecal contamination. Transcriptome analysis was performed to gain further insight into the metabolic pathways required for persistence and growth of EHEC in the bovine intestine. Understanding the physiology of EHEC in the gut of ruminants is critical to identifying the potential nutritional basis to limiting EHEC shedding. A global transcriptome analysis was performed to gain further insight into the metabolic pathways required for persistence and growth of EHEC in the bovine intestine. DNA microarrays were performed using RNA from EHEC O157:H7 EDL933 incubated in bovine small intestine content (BSIC) compared with cells incubated in M9-minimal media.

ORGANISM(S): Escherichia coli

PROVIDER: GSE49468 | GEO | 2013/08/01

SECONDARY ACCESSION(S): PRJNA213982

REPOSITORIES: GEO

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EDL933 cultured in bovine small intestine content (BSIC) vs M9 minimal media

Project description:Enterohemorrhagic Escherichia coli (EHEC) are transmitted from cattle to human by means of contaminated food products resulting from fecal contamination. Transcriptome analysis was performed to gain further insight into the metabolic pathways required for persistence and growth of EHEC in the bovine intestine. Understanding the physiology of EHEC in the gut of ruminants is critical to identifying the potential nutritional basis to limiting EHEC shedding. A global transcriptome analysis was performed to gain further insight into the metabolic pathways required for persistence and growth of EHEC in the bovine intestine. DNA microarrays were performed using RNA from EHEC O157:H7 EDL933 incubated in bovine small intestine content (BSIC) compared with cells incubated in M9-minimal media. Four biological replicates collected for bacterial cultures on separate days for each media and labelled following a dye-switch design : For each media two replicates labeled in Cy3 and two replicates in Cy5.

2013-08-01 | E-GEOD-49468 | biostudies-arrayexpress

RNA-seq to profile the H-NS transcriptome in EHEC O157:H7 strain EDL933

Project description:In order to define the role of H-NS in regulating gene transcription and further find out the biological significance of this protein in EHEC, we conducted RNA-seq and then analyzed the transcriptome data, using EHEC O157:H7 strain EDL933 and Δhns. A total of 983 genes were found to be regulated by H-NS. 213 and 770 genes exhibited lower and higher transcript levels in Δhns than in WT, respectively. For instance, chemotaxis and flagellar associated genes were down-regulated in Δhns. Besides, 34 genes on virulence plasmid pO157 were down-regulated by H-NS. The outcome of RNA-seq were verified by real time quantitative PCR. As reported in Salmonella Typhimurium, Δhns showed a growth deficiency and altered fitness. We first detected that both stx1 and stx2 in EDL933 were repressed by H-NS. Although no survival difference between EDL933 and Δhns was detected when phagocytized by macrophage, we characterized the higher ability of colonization and in consequence the higher virulence of Δhns to BALB/c mice by experimental analyses than those of WT, especially when intact commensal flora of mice existed. This might indeed help us understand the core role of H-NS in depth.

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Project description:Two lineages of enterohemorrhagic (EHEC) Escherichia coli O157:H7 (EDL933, Stx1+ and Stx2+) and 86-24 (Stx2+) were investigated in regards to biofilm formation on an abiotic surface. Strikingly, EDL933 strain formed a robust biofilm while 86-24 strain formed no biofilm on either a polystyrene plate or a polyethylene tube. To identify the genetic mechanisms of different biofilm formation in two EHEC strains, DNA microarrays were first performed and phenotypic assays were followed. In the comparison of the EDL933 strain versus 86-24 strain, genes (csgBAC and csgDEFG) involved in curli biosynthesis were significantly induced while genes (trpLEDCB and mtr) involved in indole signaling were repressed. Additionally, a dozen of phage genes were differentially present between two strains. Curli assays using a Congo red plate and scanning electron microscopy corroborate the microarray data as the EDL 933 strain produces a large amount of curli, while 86-24 forms much less curli. Also, the indole production in the EDL933 was 2-times lower than that of 86-24. It was known that curli formation positively regulates and indole negatively regulates biofilm formation of EHEC. Hence, it appears that less curli formation and high indole production in the 86-24 strain are majorly responsible for no biofilm formation.

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