Identification of potential therapeutic targets for Burkholderia cenocepacia by comparative transcriptomics
ABSTRACT:  Transcription profiling of one Burkholderia cenocepacia clinical isolate, J2315, versus a soil isolate, HI2424, in conditions mimicking CF sputum  Transcription profiling of Burkholderia cenocepacia isolates J2315 and HI2424 in media mimicking CF sputum or the soil environment  J2315 vs. HI2424 cells in the same condition.  Two-condition experiment. Biological replicates: 4 replicates.
Burkholderia cenocepacia is an endemic soil dweller and emerging opportunistic pathogen in patients with cystic fibrosis (CF). The identification of virulence factors and potential therapeutic targets has been hampered by the genomic diversity within the species as many factors are not shared among the pathogenic members of the species.In this study, global identification of putative virulence factors was performed by analyzing the transcriptome of two related strains of B. cenocepacia (one clin ...[more]
Project description: Transcription profiling of one Burkholderia cenocepacia clinical isolate, J2315, versus a soil isolate, HI2424, in conditions mimicking CF sputum  Transcription profiling of Burkholderia cenocepacia isolates J2315 and HI2424 in media mimicking CF sputum or the soil environment Overall design:  J2315 vs. HI2424 cells in the same condition.  Two-condition experiment. Biological replicates: 4 replicates.
Project description:Fang2011 - Genome-scale metabolic network of
Burkholderia cenocepacia (iKF1028)
This model is described in the article:
Exploring the metabolic
network of the epidemic pathogen Burkholderia cenocepacia J2315
via genome-scale reconstruction.
Fang K, Zhao H, Sun C, Lam CM, Chang
S, Zhang K, Panda G, Godinho M, Martins dos Santos VA, Wang
BMC Syst Biol 2011; 5: 83
BACKGROUND: Burkholderia cenocepacia is a threatening
nosocomial epidemic pathogen in patients with cystic fibrosis
(CF) or a compromised immune system. Its high level of
antibiotic resistance is an increasing concern in treatments
against its infection. Strain B. cenocepacia J2315 is the most
infectious isolate from CF patients. There is a strong demand
to reconstruct a genome-scale metabolic network of B.
cenocepacia J2315 to systematically analyze its metabolic
capabilities and its virulence traits, and to search for
potential clinical therapy targets. RESULTS: We reconstructed
the genome-scale metabolic network of B. cenocepacia J2315. An
iterative reconstruction process led to the establishment of a
robust model, iKF1028, which accounts for 1,028 genes, 859
internal reactions, and 834 metabolites. The model iKF1028
captures important metabolic capabilities of B. cenocepacia
J2315 with a particular focus on the biosyntheses of key
metabolic virulence factors to assist in understanding the
mechanism of disease infection and identifying potential drug
targets. The model was tested through BIOLOG assays. Based on
the model, the genome annotation of B. cenocepacia J2315 was
refined and 24 genes were properly re-annotated. Gene and
enzyme essentiality were analyzed to provide further insights
into the genome function and architecture. A total of 45
essential enzymes were identified as potential therapeutic
targets. CONCLUSIONS: As the first genome-scale metabolic
network of B. cenocepacia J2315, iKF1028 allows a systematic
study of the metabolic properties of B. cenocepacia and its key
metabolic virulence factors affecting the CF community. The
model can be used as a discovery tool to design novel drugs
against diseases caused by this notorious pathogen.
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Project description:In order to unravel the molecular mechanisms involved in the resistance of biofilm-grown <br>Burkholderia cenocepacia cells against high concentrations of disinfectants, the present <br>study focussed on the transcriptional response in sessile B. cenocepacia J2315 cells<br> following exposure to high levels of H2O2, NaOCl or chlorhexidine. In addition differences<br> in gene expression were examined between untreated B. cenocepacia sessile cells and <br>B. cenocepacia planktonic cells.
Project description:comparative genomic hybridization between Burkholderia cepacia sequenced strains to determine what are the limits to strain relativity in order to still provide accurate estimations of gene content Overall design: Two color experiment, Burkholderia cenocepacia J2315 (reference), B. cenocepacia AU1054 (tester), HI2424 (tester), B. ambifaria AMMD (tester), B. vietnamiensis G4 (tester), B. xenovorans LB400 (tester): At least two replicates including a single dye swap for each reference-tester comparison
Project description:Global gene expression in three clinical isolates of the ET12 lineage of Burkholderia cenocepacia was compared by microarray analysis: Strain J2315 (isolated 1989), and strains BCC1616 (HI4277) and BCC1617 (HI4283), both isolated in 2008.
Project description:Burkholderia cenocepacia J2315 was grown at 37 degrees centigrade in LB broth to the beginning of stationary phase (18 hours incubation time). <br>The expression profile was compared to cultures harvested in mid-log phase (7 hours incubation time).<br>
Project description:Hfq proteins are RNA chaperones that play a critical role in post-transcription regulation of gene expression. Bacteria of the Burkholderia cepacia complex harbor two distinct and functional Hfq proteins, the Hfq and Hfq2. We have previously performed the functional analysis of Hfq and Hfq2 in the pathogen Burkholderia cenocepacia J2315. In order to examine the impacts of each RNA chaperone on the global transcriptome of B. cenocepacia J2315, we performed comparative transcriptome profile of mutants on the hfq and hfq2 genes, using as reference the wild-type strain. For expression profiling, over-night cultures of the Burkholderia cenocepacia J2315 wild-type strain and the isogenic mutants hfq::Tp and Δhfq2 grown in LB medium were diluted to an initial OD640 nm of 0.25 into LB medium. Triplicate samples were cultured at 37ºC with 250 r.p.m. agitation for 16 h and RNA extracted from the three bacterial isolates.