Global transcriptional response of Bordetella bronchiseptica after exposure to serum
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ABSTRACT: B. bronchiseptica strain RB50 was grown in Stainer-Scholte (SS) broth at 37°C with shaking (220 rpm) overnight. Bacteria were subcultured at a starting OD600 of 0.1 into SS broth and grown at 37°C with shaking (220 rpm) up to an OD600 of 1.0. The cultures were pelleted and bacteria were washed twice with PBS buffer and equal aliquots were resuspended in SS broth (control) or in 100% sheep serum and incubated for 1 hour at 37°C.
Project description:B. bronchiseptica strain RB50 was grown in Stainer-Scholte (SS) broth at 37°C with shaking (220 rpm) overnight. Bacteria were subcultured at a starting OD600 of 0.1 into SS broth and grown at 37°C with shaking (220 rpm) up to an OD600 of 1.0. The cultures were pelleted and bacteria were washed twice with PBS buffer and equal aliquots were resuspended in SS broth (control) or in 100% sheep blood and incubated for 1 hour at 37°C.
Project description:A Bordetella bronchiseptica Bvg- phase-locked mutant (delta_bvgS) was grown in Stainer-Scholte (SS) broth at 37°C with shaking (220 rpm) overnight. Bacteria were subcultured at a starting OD600 of 0.1 into SS broth and grown at 37°C with shaking (220 rpm) up to an OD600 of 1.0. The cultures were pelleted and bacteria were washed twice with PBS buffer and equal aliquots were resuspended in SS broth (control) or in 100% sheep blood and incubated for 1 hour at 37°C.
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:B. bronchiseptica strains RB50 and 1289 were grown in SS broth, subcultured at a starting OD600 of 0.02 into 50mL of SS broth, grown at 37C for 24 hours while shaking and harvested in log phase (OD600 1.0). Total RNA was extracted with Trizol (Invitrogen, Carlsbad, CA), treated with RNase-free DNase I (Invitrogen, Carlsbad, CA) and purified using RNeasy columns (Qiagen, Valencia, CA) according to the manufacturers instructions. RNA was isolated from two independent biological replicates of strains RB50 and 1289. A 2-color hybridization format was used and dye-swap experiments were performed. For each reaction, 5ug of cDNA was fluorescently labeled. The two differentially labeled reactions to be compared were combined and hybridized to a B. bronchiseptica strain RB50 specific long-oligonucleotide microarray.
Project description:We conducted paired-end RNA sequencing on Saccharomyces cerevisiae (BY4741) to study the effects of different thermal pulsing conditions. The cells were divided into two groups: thermal pulsing (TP) and steady-state (SS) conditions. In the TP condition, cells were initially incubated at 30°C in YPD broth for 15 minutes, followed by 15 minutes at 37°C, constituting a single cycle of thermal pulsing. In the SS condition, cells were maintained at 30°C for both 15-minute intervals. This process was repeated for twenty-five cycles to produce both thermally pulsed cells and steady-state cells.
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:Using a B. bronchiseptica-specific microarray, we characterized the global gene expression profiles of several B. bronchiseptica strains, along with their phase-locked derivatives, grown at 23oC or 37oC.
Project description:B. bronchiseptica strain KM22, a virulent swine isolate, and strains TN27 and TN28, fhaB and prn deletion mutants of KM22, respectively, were cultured in Stainer-Scholte broth and grown at 37C with shaking at 275 rpm until an OD600 of 0.8 was reached. RNA extracted from KM22 was then compared to RNA extracted from TN27 and TN28.
Project description:Whole genome transcriptome analysis was performed to determine which genes were differentially expressed between strains T44625 and RB50
Project description:To identify the regulated by the transcriptional regulator BpsR, whole genome transcriptome analysis performed to determine which genes were differentially expressed between an in-frame bpsR deletion-mutant of B. bronchiseptica and its wild-type parental strain RB50.