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.
Project description:[1] Transcription profiling of one Burkholderia cenocepacia clinical isolate, J2315, versus a soil isolate, HI2424, in conditions mimicking CF sputum [2] Transcription profiling of Burkholderia cenocepacia isolates J2315 and HI2424 in media mimicking CF sputum or the soil environment
Project description:To determine whether CRISPR interference can be used to recapitulate a glycosylation null mutant strain in Burkholderia cenocepacia via data independent acquisition mass spectrometry
Project description:Burkholderia cenocepacia is a versatile opportunistic pathogen that survives in a wide variety of environments, which can be limited in nutrients such as nitrogen. We previously showed that B. cenocepacia sigma factor s54 played a major role in control of nitrogen assimilation and virulence. In this work, we investigated the role of the s54 enhancer binding protein NtrC in controlling the response to nitrogen limitation and virulence. RNA-Seq analyses and phenotypical analysis on a ntrC mutant strain showed that, in addition to orchestrating uptake of nitrogen sources, NtrC is also regulating exopolysaccharide (EPS) production and motility. A search for NtrC consensus sequences identified a potential binding sequence in the promoter region of gene clusters involved in EPS formation and flagellar rotation suggesting that NtrC directly controls the expression of these phenotypic traits in B. cenocepacia H111.
Project description:Burkholderia cenocepacia sequence type 32 (ST32) represents one of the most globally distributed strains from Bukrholderia cepacia complex (Bcc), which infected 30% of Czech cystic fibrosis (CF) patients. The aim of this study was to compare gene expression in two pairs of ST32 clinical isolates that were subjected to cultivation in two different conditions, characteristic for chronic B. cenocepacia infection in CF patients. ST32 strain is known to be a problematic epidemic strain, which caused a serious outbreak at the Prague CF centre.
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 M-NM-^Thfq2 grown in LB medium were diluted to an initial OD640 nm of 0.25 into LB medium. Triplicate samples were cultured at 37M-BM-:C with 250 r.p.m. agitation for 16 h and RNA extracted from the three bacterial isolates.
