Gene Expression Changes in Campylobacter jejuni triggered by exposure to Novobiocin
Ontology highlight
ABSTRACT:
ORGANISM(S): Campylobacter jejuni
SUBMITTER: Yang Wang
PROVIDER: E-GEOD-18415 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:In order to investigate the gene experssion changes in Campylobacter jejuni triggered by exposure to Novobiocin.
2009-12-28 | GSE18415 | GEO
Project description:Campylobacter jejuni is a major zoonotic pathogen transmitted to humans via the food chain. C. jejuni is prevalent in chickens, a natural reservoir for this pathogenic organism. Due to the importance of macrolide antibiotics in clinical therapy of human campylobacteriosis, development of macrolide resistance in Campylobacter has become a concern for public health.To facilitate understanding the molecular basis associated with the fitness difference between Erys and Eryr Campylobacter, we compared the transcriptomes between ATCC 700819 and its isogenic Eryr transformant T.L.101 using DNA microarray. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni. Four hybridizations were performed each with independently extracted samples of either macrolide susceptible ATCC 700819 cDNA samples or its isogenic Eryr transformant T.L.101 cDNA samples. A dye swap was utilized to help minimize dye dependent bias. Thus, there were four biological replicates of each sample.
2011-11-02 | E-GEOD-33330 | biostudies-arrayexpress
Project description:Objectives: The aim of the study was to characterize (-)-α-Pinene, which is one of the chemical constituents of Alpinia katsumadai seed essential oil responsible for its resistance modulatory activity in Campylobacter jejuni. Methods: Broth microdilution method was used to evaluate the antimicrobial and resistance modulatory potential of (-)-α-Pinene and ethidium bromide accumulation assay to determine its efflux-inhibitory activity in subinhibitory concentration. The target efflux system was identified using knock-out mutants in several efflux related genes. Furthermore, the influence of subinhibitory concentration of (-)-α-Pinene on C. jejuni NCTC 11168 was investigated using microarray technology in order to elucidate the adaptive mechanism of bacteria to treatment with this phytochemical. Knock-out mutants of key adaptation genes were constructed and their role in adaptation to several stress factors, including (-)-α-Pinene, different osmolites and pH, was investigated using Biolog phenotypical microarrays and CFU counts. Results: (-)-α-Pinene was confirmed as highly efficient Campylobacter jejuni resistance modulator, due to its efflux inhibitory activity, which was significantly higher compared to reference inhibitors CCCP and reserpine. The CmeABC along with newly characterized CmeI (Cj1687) was confirmed as its main target efflux system. The transcriptional analysis indicated that the heat shock regulators HspR and HrcA are the main transcription regulators involved in adaptation to (-)-α-Pinene treatment. Conclusions: (-)-α-Pinene is a novel CmeABC and CmeI efflux inhibitor, which evokes the heat shock response in Campylobacter jejuni. Influence of (-)-α-Pinene on gene expression in C. jejuni NCTC 11168 was determined by expressional analysis and qRT-PCR. Exponential phase culture was adjusted to OD600=0.2 in MHB using spectrophotometer (Smart Spec, Bio-Rad, Hercules, CA, USA). Five ml of culture was treated with 62.5 mg/L of (-)-alpha-pinene dissolved in DMSO. Only DMSO (0.048 %) was added to untreated samples. Cultures were treated for 2 h, incubated microaerobically shaking (160 rpm) at 42°C. Experiments were carried out in 4 biological replicates. RNA Protect Bacteria reagent (Qiagen, Maryland, USA) was added to the culture and total RNA was isolated using RNeasy mini kit (Qiagen) and treated with Ambion® Turbo DNA-freeTM kit (Invitrogen, USA). Microarrays with 4751 probes targeting 1756 genes specific for Campylobacter jejuni subsp. jejuni NCTC 11168 (Mycroarray, Biodiscovery-LLC, MI, USA) were used for gene expression analysis. The cDNA was synthesized with random hexamers, SuperScriptTM III Reverse Transcriptase and amynoallyl dUTP (all supplied by Invitrogen, USA) and labeled with monofunctional NHS-ester dye Amersham C3 or Cy5 (GE Healthcare, Buckinghamshire, UK). Concentration of cDNA and labeling efficiency was determined spectrophotometrically with NanoDrop 1000. Four biological replicates were hybridized to four microarrays according to manufacturerâs protocol, incubated for 24 hours at 42â°C and scanned at 532-nm (Cy3) and 635-nm (Cy5) wavelengths using GenePix 4100A (Molecular Devices, Sunnyvale, CA) following the manufacturer's protocol. Fluorescence intensities of each spot were extracted using GenePix Pro 7.0 (Molecular Devices).
2015-11-19 | E-GEOD-59879 | biostudies-arrayexpress
Project description:Campylobacter, a major foodborne pathogen, is increasingly resistant to macrolide antibibotics. previous findings suggeted that development of macrolide resistance in campylobacter requires a multi-step process, but the molecular mechanisms involved in the process are not known. In our study, multiple seriess of macrolide-resistant C. jejuni mutants were selected in vitro by stepwise exposure of C. jejuni 81-176 to increasing concentrations of erythromycin and tylosin. A set of the selected resistance were subjected to microarray and the the global transcriptional profile was analyzed. In this sery, DNA microarray was used to compare the gene expression profiles of macrolide resistant strains (76E2, 76E8 and 76E64) with its parent wild-type strain C. jejuni 81-176. This assay identified a large number of genes that showed >1.5 fold changes (q-value<0.1) in expression in the macrolide resistant strains. The up-regulated genes are involved in surface structure,ribosomal, heat shock and some specific Misc memerbrane , while the majority of the down-regulated genes are involved in energy metabolism, amino acid biosynthesis. The over-expression of genes involved in surface structure and Misc memerbrance was associated with the development of intermedial-level resistance to macrolide in campylobacter 81-176. Keywords:intermedial-level macrolide resistant C. jejuni selected from C. jejuni 81-176. step-wise selection. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni. Four hybridizations were performed each with independently extracted samples of either macrolide susceptible C. jejuni 81-176 cDNA samples or macrolide resistant C. jejuni cDNA samples. A dye swap was utilized to help minimize dye dependent bias. Thus there were three to four biological replicates of each sample.
2010-12-28 | E-GEOD-17899 | biostudies-arrayexpress
Project description:Erythromycin is the drug of choice to treat campylobacteriosis, but resistance to this antibiotic is rising. The adaptive mechanisms employed by Campylobacter jejuni to erythromycin treatment remain unknown. The aim of this study is to determine the molecular basis underlying CampylobacterM-bM-^@M-^Ys immediate response to Ery treatment. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni macrolide resistant strain JL272. One hybridizations were performed: sham-treated JL272 v.s. lethal dose erythromycin treated JL272. Samples were independently grown and harvested. There were three biological replicates of each sample.
2013-10-10 | E-GEOD-43025 | biostudies-arrayexpress
Project description:Erythromycin is the drug of choice to treat campylobacteriosis, but resistance to this antibiotic is rising. The adaptive mechanisms employed by Campylobacter jejuni to erythromycin treatment remain unknown. The aim of this study is to determine the molecular basis underlying CampylobacterM-bM-^@M-^Ys immediate response to Ery treatment. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni wild type strain NCTC 11168. One hybridizations were performed: sham-treated NCTC 11168 v.s. lethal dose erythromycin treated NCTC 11168. Samples were independently grown and harvested. There were three biological replicates of each sample.
2013-10-10 | E-GEOD-43023 | biostudies-arrayexpress
Project description:Erythromycin is the drug of choice to treat campylobacteriosis, but resistance to this antibiotic is rising. The adaptive mechanisms employed by Campylobacter jejuni to erythromycin treatment remain unknown. The aim of this study is to determine the molecular basis underlying CampylobacterM-bM-^@M-^Ys immediate response to Ery treatment. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni wild type strain NCTC 11168. One hybridizations were performed: sham-treated NCTC 11168 v.s. sub-lethal dose erythromycin treated NCTC 11168. Samples were independently grown and harvested. There were three biological replicates of each sample.
2013-10-10 | E-GEOD-43024 | biostudies-arrayexpress
Project description:Campylobacter, a major foodborne pathogen, is increasingly resistant to macrolide antibibotics. Previous findings suggeted that development of macrolide resistance in campylobacter requires a multi-step process, but the molecular mechanisms involved in the process are not known. In our study, multiple series of macrolide-resistant C. jejuni mutants were selected in vitro by stepwise exposure of C. jejuni NCTC11168 to increasing concentrations of erythromycin and tylosin. A set of the selected resistance were subjected to microarray and the the global transcriptional profile was analyzed. In this sery, DNA microarray was used to compare the gene expression profiles of macrolide resistant strains (68E1, 68E8 and 68E64) with its parent wild-type strain NCTC11168. The assay identified a small number of genes that showed significant changes (q-value<0.1) in expression in the low-level macrolide resistant strain 68E1, while a large number of gene showing significant changes in intermedia-level resistant stran 68E8 and high-level resistant strain 68 E64. The up-regulated genes in the resistant strains are involved in miscellaneous periplasmic proteins, efflux protienand putative aminotransferase, while the majority of the down-regulated genes are involved in electron transport,lipoprotein, heat shock protein and unknown function proteins. These findings suggest that there is not much change in low-level macrolide resistant C. jejuni strain. The over-expression of efflux pump and periplasmic protein was involved in the development of resistance to macrolide in C. jejuni. Keywords: macrolide resistant C. jejuni selected from NCTC 11168 step-wise selection. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni. Four hybridizations were performed each with independently extracted samples of either macrolide susceptible C. jejuni NCTC11168 cDNA samples or macrolide resistant C.jejuni cDNA samples. A dye swap was utilized to help minimize dye dependent bias. Thus, there were four biological replicates of each sample.
2010-12-28 | E-GEOD-17881 | biostudies-arrayexpress
Project description:DksA is well-known for its regulatory role in the transcription of ribosomal RNA and genes involved in amino acid synthesis in many bacteria. DksA is also reported to control expression of virulence genes in pathogenic bacteria. Here, we elucidated the roles of the DksA-like protein (CJJ81176_0160, Cj0125c) in the pathogenesis of Campylobacter jejuni. Like in other bacteria, transcription of stable RNA was repressed by DksA under stressful conditions in C. jejuni. Transcriptomic and proteomic analyses of C. jejuni 81-176 and its isogenic dksA mutant showed differential expression of many genes involved in iron-related metabolism, flagellar synthesis and amino acid metabolism. Also the dksA mutant of C. jejuni demonstrated a decreased ability to invade into intestinal cells and to induce release of interleukin-8 from intestinal cells. These results suggest the DksA-like protein plays an important regulatory role in the physiology and virulence of C. jejuni. Keywords: dksA mutation of Campylobacter jejuni The design utilized a commercially available two color microarray slide for the enture transcriptome of Campylobacter jejuni. Six hybridizations were performed each with independently extracted samples of either C. jejuni wildtype of C. jejuni dksA mutant cDNA samples. A dye swap was utilized to help minimize dye dependent bias. Thus there were six biological replicates of each sample.
2008-05-05 | E-GEOD-9866 | biostudies-arrayexpress
Project description:Campylobacter jejuni (C. jejuni) protein microarrays were used to identify immunogenic C. jejuni proteins that may be useful in the development of biomarkers, diagnostic assays, or subunit vaccines for humans or livestock. A native protein microarray with over 1400 individually purified GST-tagged C. jejuni proteins (86 % of the proteome), was constructed and screened for antibody titers present in test sera. The protein arrays were screened with antisera from rabbits inoculated with whole C. jejuni or various serotypes of E. coli or Salmonella, with antisera from mice infected with C. jejuni, and sera from healthy humans. Dual detection of GST signals was incorporated as a way of normalizing the variation of protein concentrations contributing to the antibody staining intensities.
2018-12-29 | E-MTAB-7535 | biostudies-arrayexpress