Table 1.
Bacterial strains, plasmids and primers used in this study
| Strains or plasmid | Description | Reference |
|---|---|---|
| E. coli | ||
| DH5⟨ | F2ø80dlacZ⊗M15 endA1 recA1 gyrA96 thi-1 hsdR17(rK- mK+) supE44 relA1 deoR Δ(lacZYA-argF)U169 | [56] |
| H1717 | aroB fhuF::λplacMu | [40] |
| H1717 (pFur616) | E. coli H1717 carrying pFur616 | This study |
| H1717 (pFur616-kanP) | E. coli H1717 carrying pFur616-kanP | This study |
| H1717 (pFur616-kanC) | E. coli H1717 carrying pFur616-kanC | This study |
| H1780 | araD139∆aargF-lacU169rpsL150 relA1 flbB5301deoC1 ptsF25 rbsR fiu::lacZ fusion lacking Fur | [40] |
| H1780 (pFur616) | E. coli H1780 carrying pFur616 | This study |
| H1780 (pFur616-kanP) | E. coli H1780 carrying pFur616-kanP | This study |
| H1780 (pFur616-kanC) | E. coli H1780 carrying pFur616-kanC | This study |
| H1780 (pFur730) | E. coli H1780 carrying pFur730 | This study |
| H1780 (pFur1722) | E. coli H1780 carrying pFur1722 | This study |
| N. europaea | ||
| ATCC 19178 | Wild type | ATCC |
| fur:kanP | Insertion of kan cassette in the furbox upstream of NE0616 gene | This study |
| Plasmids | ||
| pGEM-T Easy | Vector for cloning PCR products; Ampr | Promega |
| pFur616 | pGEM-T Easy vector containing NE0616 u&d* region | This study |
| pFur616-kanP | In vitro transposon mutagenesis of pFur616 with EZ-Tn5 <KAN-2> with kan cassette insertion in fur box located in promoter region of NE0616 | This study |
| pFur616-kanC | In vitro transposon mutagenesis of pFur616 with EZ-Tn5 <KAN-2> with kan cassette insertion in C-terminal region of NE0616 | This study |
| pFur730 | pGEM-T Easy vector containing NE0730 u&d region | This study |
| pFur1722 | pGEM-T Easy vector containing NE1722 u&d region | This study |
| Primers used for cloning, mutagenesis and mutant confirmation | ||
| NE0616u&d-1 | 5'-ATCCTGGAAGAAAACGGTCA-3' | This study |
| NE0616u&d-2 | 5'-TGCAGGTTTCAAACGAAAAA-3' | This study |
| NE0730u&d-1 | 5'-TTTCAGACGTTGCTGACAAAA-3' | This study |
| NE0730u&d-2 | 5'-TCATTTTGGCTGTTCATTTCA-3' | This study |
| NE1722u&d-1 | 5'-TATGGCTTACGGAAAACGGTA-3' | This study |
| NE1722u&d-2 | 5'-ACAAAAACAGACACGGAGGAA-3' | This study |
*- u&d denotes upstream & downstream region.