TABLE 1.
Strains, plasmids, and primers
| Strain, plasmid, or primer | Genotype description or sequencea | Reference or source |
|---|---|---|
| Strains | ||
| A. caldus MTH-04 | Isolated from Tengchong, Yunnan Province, China | Lab stock |
| A. caldus MTH-04 Δfur | fur gene deletion | This study |
| A. caldus MTH-04 Δfur(fur) | fur gene complementation for Δfur | This study |
| A. caldus MTH-04 Δfur(SI) | fur gene complementation for Δfur; H31A H88A of fur | This study |
| A. caldus MTH-04 Δfur(SII) | fur gene complementation for Δfur; E106A H123A of fur | This study |
| A. caldus MTH-04 Δfur(SI&II) | fur gene complementation for Δfur; H31A H88A E106A H123A of fur | This study |
| A. caldus MTH-04 Δfur-dC | fur gene complementation for Δfur; deletion of C-terminal extension of fur | This study |
| E. coli DH5α | F− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 end A1 recA1 hsdR17(rK− mK+) supE44λ-thi-1 gyr96 relA1 phoA | TransGen Biotech |
| E. coli SM10 | thr leu hsd recA Kmr RP4–2-Tc::Mu | 61 |
| E. coli BL21(DE3) | F− ompT hdsSB(rB− mB−) gal dgmmet (DE3) | TransGen Biotech |
| Plasmids | ||
| pSDUDI | Ampr Kmr; oriTRP4 | Lab stock |
| pMSD1-I-Sce I | Smr; mob+; I-SceI gene | Lab stock |
| pSDUDI∷fur (UHA+DHA) | Suicide plasmid for fur deletion | This study |
| pSDUDI∷fur (UHA+fur+DHA) | Suicide plasmid for fur complementation | This study |
| pSDUDI∷fur (UHA+fur(SI)+DHA) | Suicide plasmid for fur complementation; H31A H88A of fur | This study |
| pSDUDI∷fur (UHA+fur(SII)+DHA) | Suicide plasmid for fur complementation; E106A H123A of fur | This study |
| pSDUDI∷fur (UHA+fur(SI&II)+DHA) | Suicide plasmid for fur complementation; H31A H88A E106A H123A of fur | This study |
| pSDUDI∷fur (UHA+fur-dC+DHA) | Suicide plasmid for fur complementation; deletion of C-terminal extension of fur | This study |
| pJRD215-Luc-Cm | Smr Kmr; IncQ Mob+; luc cat | Lab stock |
| pJRD215-Pfur-Luc-Cm | Smr Kmr; IncQ Mob+; luc cat; Pfur | This study |
| pET-22b | Ampr | Novagen |
| pET-22b-Fur | Ampr; fur | This study |
| pET-22b-Fur(SI) | Ampr; H31A H88A of fur | This study |
| pET-22b-Fur(SII) | Ampr; E106A H123A of fur | This study |
| pET-22b-Fur(SI&II) | Ampr; H31A H88A E106A H123A of fur | This study |
| pET-22b-dC | Ampr; deletion of C-terminal extension of fur | This study |
| Primers | ||
| fur UF-XbaI | TTCTAGGCTCTAGAGACAGGGAGCAGGAACG | This study |
| fur UR-SpeI | TTCTAGACTAGTCGGGTCGCGCACACCC | This study |
| fur DF-SpeI | TTCTAGACTAGTGAAACTCATGGCCAAAGTCTGA | This study |
| fur DR-HindIII | TTTCCCAAGCTTGTGGGTTCTGCCAATCTC | This study |
| F1 | TCACGATTTGACCGAGCC | This study |
| R1 | CCTCAAGGCCACGCTC | This study |
| F2 | TTCAGCCTGGGTCTCG | This study |
| R2 | CATTGGTCGGGGTGCC | This study |
| F3 | ATCTGCATCCCCACTTCT | This study |
| R3 | CACTTTTGCGCTTTGGTA | This study |
| D-F-fur | GCCAGCCCTTTTCAATTCATCGCTGTGCATGAAACTCATGGCCAAAGTCTGA | This study |
| U-R | CGGGTCGCGCACACCC | This study |
| fur-F-U | CCTGAGGCGCTCAGGGTGTGCGCGACCCGTCACGATTTGACCGAGC | This study |
| fur-R | ATGCACAGCGATGAATTG | This study |
| fH31A F | ACCAGCGAGAGCCGCGCTCTCACCGCCGAGGAG | This study |
| fH31A R | CTCCTCGGCGGTGAGAGCGCGGCTCTCGCTGGT | This study |
| fH88A F | AGCGGCCACCACGATGCCATGGTCTGCACCGCC | This study |
| fH88A R | GGCGGTGCAGACCATGGCATCGTGGTGGCCGCT | This study |
| fE106A F | TACGACGAGATCCTGGCCGCACGCCAGCAAAGC | This study |
| fE106A R | GCTTTGCTGGCGTGCGGCCAGGATCTCGTCGTA | This study |
| fH123A F | TTTCATCTCTCGGACGCTAGCCTCTATCTCTAC | This study |
| fH123A R | GTAGAGATAGAGGCTAGCGTCCGAGAGATGAAA | This study |
| Pfur-F-MluI | CGACGCGT ATCGTCCTTGAACTGCA | This study |
| Pfur-R-NdeI | GGAATTCCATATGGAAACTCATGGCCAAAG | This study |
| Fur F NdeI | GGAATTCCATATGATGCACAGCGATGAATTG | This study |
| Fur R XhoI | CCCTCGAGCGATTTGACCGAGC | This study |
| dFur R XhoI | CCCTCGAGTCAGGTTCCGTAGAGATAGA | This study |
| FeoABC-F | GCGTTTTACACCCAGGCAC | This study |
| FeoABC-R | TTCCACCTGCTCACCTGC | This study |
| FeoP-F | GCGTAGGACGTCCTTGATA | This study |
| FeoP-R | GCTGGAAGCAAGCATGGTG | This study |
| feoAF | CGCCGTCCTTCAAATCG | This study |
| feoAR | GTCATCCTGCCTGTTCCC | This study |
| feoPF | GGGATCCAAGCTCGGTATAT | This study |
| feoPR | CCACTTTCACGACATAGCCA | This study |
a
Underlining indicates restriction endonuclease sites.