Construction and validation of a first-generation Bordetella bronchiseptica long-oligonucleotide microarray by transcriptional profiling the Bvg regulon

• Published in: BMC genomics Vol. 8; no. 1; p. 220
• Main Author: Nicholson, Tracy L
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 06-07-2007; BioMed Central; BMC
• Subjects: Bordetella bronchiseptica; Bordetella bronchiseptica - genetics; Cluster Analysis; DNA microarrays; Gene Expression Profiling - methods; Gene Expression Regulation, Bacterial; Genes, Bacterial; Genetic aspects; Genetic transcription; Gram-negative bacteria; Oligonucleotide Array Sequence Analysis - methods; Regulon; United States
• ISSN: 1471-2164; 1471-2164
• DOI: 10.1186/1471-2164-8-220

Bordetella bronchiseptica is a bacterial respiratory pathogen that infects a broad range of mammals, causing chronic and often subclinical infections. Gene expression in Bordetella is regulated by a two-component sensory transduction system, BvgAS, which controls the expression of a spectrum of phenotypic phases transitioning between a virulent (Bvg+) phase and a non-virulent (Bvg-) phase.
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. Data from this microarray analysis revealed 1,668 Bvg-regulated genes, which greatly expands the BvgAS regulon defined in previous reports. For previously reported Bvg-regulated transcripts, the gene expression data presented here is congruent with prior findings. Additionally, quantitative real-time PCR data provided an independent verification of the microarray expression values.
The results presented here provide a comprehensive, genome-wide portrait of transcripts encompassing the BvgAS regulon, while also providing data validating the long-oligonucleotide microarray described here for studying gene expression in Bordetella bronchiseptica.