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0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 0 10 20 30 40 C ycle num ber Fluorescence (530 nm ) Δ arrF WT WT ∆arrF gure 2. Confirmation of complete knockout of arrF gene in ∆arrF mutant using real-tim curve representing the cycle-dependent fluorescence associated with amplification of a A samples isolated from wild type and ∆arrF mutant. A primers-specific DNA was amplifie type strain with the cross point (CP) value of ca. 17, whereas the DNA amplification w ant even after 40 cycles of amplification (inset). In the same conditions, the expres l omlA was little affected by arrF mutation. These results strongly indicate that the ar ted in the mutant and that, unlike Pseudomonas spp, A. vinelandii does not have other copy l to the arrF.
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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 - PowerPoint PPT Presentation
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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 ofinternal 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 isnearly identical to the arrF.
