Project description:The classical bordetellae (Bordetella pertussis, B. parapertussis, and B. bronchiseptica) are obligate aerobes that use only oxygen as their terminal electron acceptor for electron transport-coupled oxidative phosphorylation. Therefore, access to oxygen is critical for these bacteria to survive. To better understand how B. bronchiseptica changes its gene regulation when faced with different levels of oxygen, we grew liquid cultures of B. bronchiseptica RB50 in ambient air, 5% oxygen, and 2% oxygen. We also measured how the presence of 5% carbon dioxide affected gene expression in these bacteria, since they are respiratory pathogens and therefore get exposed to higher carbon dioxide levels during infection than are found in ambient air.
Project description:Bordetella bronchiseptica RB50 was shifted from iron replete to either iron depleted or iron replete media, and samples were taken post shift for transcriptional profiling
Project description:B. bronchiseptica RB50 was grown in medium either lacking or containing iron (ferrous sulfate). At various time points, samples were taken for gene expression analysis.
Project description:Bordetella bronchiseptica is a gram-negative respiratory pathogen that causes a diverse spectrum of respiratory disease in a wide-range of hosts. We sought to determine if strains of B. bronchiseptica differed in virulence using the mouse model of infection. Mean lethal doses (LD50) of different B. bronchiseptica strains varied widely in the murine model. B. bronchiseptica strain 253 had a LD50 that was 10-fold lower than the prototypical and fully sequenced B. bronchiseptica strain RB50. Using whole genomic transcriptome analysis covering 100% of B. bronchisetpctica strain RB50ÃÂÃÂs predicted open reading frames (ORFs), 253 was identified as lacking expression of adenylate cyclase toxin (ACT).
Project description:B. bronchiseptica strains RB50 and 1289 strains were grown in SS broth at 37°C with shaking overnight and genomic DNA was isolated from bacterial cultures using a DNA extraction kit (Qiagen, Valencia, CA) and digested with DpnII. For each labeling reaction, 2 ug of digested genomic DNA was randomly primed using Cy-5 and Cy-3 dye-labeled nucleotides, with BioPrime DNA labeling kits (Invitrogen, Carlsbad, CA) and the two differentially labeled reactions to be compared were combined and hybridized to a B. bronchiseptica RB50 specific long-oligonucleotide microarray.
Project description:Bordetella bronchiseptica is a gram-negative respiratory pathogen that causes a diverse spectrum of respiratory disease in a wide-range of hosts. We sought to determine if strains of B. bronchiseptica differed in virulence using the mouse model of infection. Mean lethal doses (LD50) of different B. bronchiseptica strains varied widely in the murine model. B. bronchiseptica strain 253 had a LD50 that was 10-fold lower than the prototypical and fully sequenced B. bronchiseptica strain RB50. Using whole genomic transcriptome analysis covering 100% of B. bronchisetpctica strain RB50ÃÂs predicted open reading frames (ORFs), 253 was identified as lacking expression of adenylate cyclase toxin (ACT). Using whole genomic comparative genomic hybridization analysis and whole genome sequencing, we determined that the cya operon, which is required for ACT production, was absent from the 253 genome.
Project description:B. bronchiseptica RB50 and a constitutive Bvg+ phase-locked mutant were grown at 37 degrees C in media containing a range of nicotinic acid concentrations.
Project description:Based on the genomic sequence and using the freely available software ArrayOligoSelector, a long oligonucleotide B. bronchiseptica microarray was designed and assembled. This long-oligonucleotide microarray was subsequently tested and validated by comparing changes in the global expression profiles between B. bronchiseptica RB50 and its Bvg- phase-locked derivative, RB54.
