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WT. ∆ arrF. WT. Δ arrF. Supplemental Figure 2. Confirmation of complete knockout of arrF gene in ∆ arrF mutant using real-time RT-PCR. Figure shows a curve representing the cycle-dependent fluorescence associated with amplification of arrF gene
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WT ∆arrF WT ΔarrF Supplemental Figure 2. Confirmation of complete knockout of arrF gene in ∆arrF mutant using real-time RT-PCR. Figure shows a curve representing the cycle-dependent fluorescence associated with amplification of arrF gene product from RNA samples isolated from wild type and ∆arrF mutant. A primers-specific DNA was amplified in the sample of wild-type strain with the cross point (CP) value of ca. 17, whereas the DNA amplification was not observed in the ∆arrF mutant even after 40 cycles of amplification (inset). In the same conditions, the expression of internal control omlA was little affected by arrF mutation. These results strongly indicate that the arrF gene is completely deleted in the mutant and that, unlike Pseudomonas spp, A. vinelandii does not have other copy that is nearly identical to the arrF.