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. pertussis Tohama I was cultivated in iron-depleted or iron-repleted medium in order to monitor proteins which are influence by iron supply. More than 200 proteins displayed increased or decreased levels. Proteins involved in iron acquisition, biofilm production and oxidative stress response were increased during iron starvation. Limited iron supply caused on the other hand decreased levels of virulence factors regulated by the BvgAS system.
Project description:comparative genomic hybridization (CGH) analysis was perfromed in order to identify coding regions present in strain T44625 that are either absent or contain a high degree of sequence divergence compared to the RB50 reference strain.
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:Microarray analysis was performed to determine if the growth-dependent gene expression changes occurred on a global scale for B. bronchiseptica strains RB50, RB53, and RB54. For this analysis cDNA representing three distinct phases of growth: mid-log phase (15H), late-log phase (24H), and late-stationary phase (48H) were directly compared.
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:Comparison of B. bronchiseptica strains RB50 and 761 grown in either atmospheric concentrations of oxygen and carbon dioxide (normal conditions) or in atmospheric levels of oxygen with the addition of 5% carbon dioxide into a sealed incubator (5% CO2 conditions).