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Figure 4 | BMC Microbiology

Figure 4

From: Role of a Fur homolog in iron metabolism in Nitrosomonas europaea

Figure 4

In vitro transposon mutagenesis scheme and mutant confirmation. (A) The physical structure of a 5,810-bp fragment of the N. europaea chromosome is shown in the center (heavy black line), with positions of NE0616 (fur) gene shown as grey arrow, the fur box (fb) located in NE0616 promoter region shown as white rectangle. The regions covered by the plasmids pFur616, pFur616-kanP, pFur616-kanC whose DNA sequences were determined are shown as thin black lines with the names of the respective plasmids shown below each line. The position and relative orientation of each in vitro-constructed Tn5-Kan2 cassette insertion mutation are indicated by a flag on the lines. The restriction endonuclease sites P (Pst1) and E (EcoR1) used for Southern blot confirmation are indicated. (B) Verification of mutagenesis of fur:kanP in N. europaea by Southern hybridization. Genomic DNA from the wild type (WT), fur:kanP mutant (MT) were digested with E (EcoRI) and P (Pst1), and probed with (left) fur ORF sequence and (right) kan sequence.

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