Figure 2 | Scientific Reports

Figure 2

From: Selection and validation of reference genes for gene expression studies in Klebsiella pneumoniae using Reverse Transcription Quantitative real-time PCR

Figure 2

Validation of the putative Fur boxes identified on the upstream region of the genes cirA, iroN and fiu by FURTA (A) and EMSA (B). In (A), Lac− indicates FURTA-negative phenotype, whereas Lac+ indicates FURTA-positive phenotype. All putative Fur boxes resulted on red E. coli H1717 colonies on MacConkey plates, which were interpreted as FURTA positive results. In (B), EMSA of the DNA fragments containing the putative Fur box of cirA, iroN, fiu and the negative control (DNA fragment without Fur box). Lanes 1, 2 and 3 contained 50 ηg of the respective DNA probes. The DNA probes were incubated with 500 ηM of His-Fur protein either in the presence of divalent cation (Lanes 2) or under divalent cation-free conditions by adding 2 mM EDTA (Lanes 3). Open arrowheads indicate the free DNA probes, while closed arrowheads indicate the mobility shift corresponding to the Fur/DNA complexes. Full-length gels are presented in Supplementary Fig. S3.

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