ABSTRACT: To investigate the role of RisX in regulating the RisA regulon, we designed and assembled a long-oligonucleotide microarray representing the full B. pertussis genome. For these experiments RNA was extracted from wild-type B. pertussis strain Bp536, ?risA, and ?risX isogenic mutants, converted to cDNA, labeled fluorescently labeled and directly compared by hybridizion to a B. pertussis 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: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: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: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.
Project description:To identify global stage-specific gene sets unique to biofilm formation for B. bronchiseptica, whole-genome transcriptome analysis was used to measure mRNA abundance under biofilm and planktonic growth conditions after 6, 12, 24, 36, and 48 hours of growth.
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.
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).
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:Whole genome transcriptome analysis was performed to determine which genes were differentially expressed between strains T44625 and RB50