S1 Fig. Schematic representation of how Pcb1692Δfur mutant strain was generated. A)
Using specific set of primers, PCR amplifications of the fur upstream and downstream regions
were generated as indicated in S1A Fig. Kanamycin cassette was amplified from pKD4 plasmid
with primers Kan F and Kan R. B) Primers Fur F and R, were used in a PCR reaction consisting
of, the fur upstream kanamycin and downstream PCR fragment to generate a PCR fusion
product. C) The fusion product was electroporated into electrocompetent Pcb1692 to generate
the Pcb1692Δfur mutant strain (S1D Fig). Both electrocompetent Pcb1692 and Pcb1692Δfur
mutant strain were electroporated with empty pTrc99A.
S2 Fig. PCR amplicons used to generate the fur mutant. Lane 1. DNA ladder, 2. fur downstream
PCR fragment, 3. kanamycin cassette PCR product, 4. fur upstream PCR fragment, 5.
Fusion product consisting of the downstream, kanamycin and upstream fragment. 6. The fragment
used for complementation which contains the fur gene and its promoter region. 7. Control.
S3 Fig. Schematic presentation of the putative promoter regions of selected genes in
Pcb1692. Based on our qRT-PCR results, some of the genes under the Pcb1692 Fur regulon
were aligned to the consensus fur box and the putative fur boxes for each gene is indicated by
red boxes.
