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. 2012 Mar;80(3):982–995. doi: 10.1128/IAI.05918-11

Fig 4.

Fig 4

Characterization of M. catarrhalis O35EΔ1550. (A) Wild-type O35E locus containing MC ORF 1550. (B) Insertion of the Kan cartridge into a deletion within MC ORF 1550 in O35EΔ1550. (C) Reverse transcriptase PCR (RT-PCR)-based analysis of transcriptional linkage between selected ORFs downstream from MC ORF 1550 in the O35E::spec strain. The primer pair (Table 2) used for each experiment is specified at the top of the figure, as is the relevant intergenic region. Lanes: 1 and 4, samples from RT-PCR with RNA as the template but no added RT; 2 and 5, samples from RT-PCR with RNA as the template and RT added; 3 and 6, samples from RT-PCR with chromosomal DNA. Size position markers (in base pairs) are present on the left. A representative experiment is shown. (D) Real-time RT-PCR analysis of relative transcript levels for MC ORF 1550, the small hypothetical ORF (hyp. ORF), and MC ORF 1549 in O35EΔ1550 relative to the surrogate wild-type O35E::spec strain. Three independently isolated sets of RNA samples (#1, #2, and #3) were used for these experiments. Statistical analysis was performed using a one-sample t test with Bonferroni's correction. (E) Growth of the O35E::spec (closed circles) and O35EΔ1550 (open circles) strains in BHI broth. These data represent the mean from two independent experiments.