M. catarrhalis Gene Expression in the Chinchilla Nasopharynx

M. catarrhalis strain O35E.rpsl was inoculated into the nasopharynx of healthy, male adult chinchillas. 24 hours later the nasopharyngeal tissues were extracted and homogenized. Total RNA was extracted from these tissue samples. Subsequently, M. catarrhalis genome directed primers were utilized to synthesize cDNA from the total RNA sample. As a control, M.catarrhalis strain O35E.rpsl was grown in BHI broth to a Klett density of 200 units and underwent RNA extraction per standard protocols. The genome directed primers mentioned above were utilized to synthesize cDNA. Both cDNA samples were subsequently labelled with either Cy3 or Cy5 and hybridized to a custom Microarrays, Inc. gene chip and scanned after 16 hours. Differential gene expression was measured utilizing the broth grown cells as the baseline and the chinchilla isolated cells as the experimental variable. There are 5 individual sample results included in this series. These represent the data from four individual biological replicates (i.e. 4 different sets of inoculated animals). For each replicate the control samples are simultaneously grown broth samples. Three dye swap experiments were performed.