Table 1.
List of strains used and engineered in this study.
| Strain | Relevant characteristic(s)1 | Source |
|---|---|---|
| A. actinomycetemcomitans | ||
| HK1651R | Clinical isolate HK1651, genome project; (R) rough colony variant | ATCC2 |
| HKR.Nal | HK1651R with a spontaneous mutation (Nalr) | This work |
| HKR.fur::kan | HKR.Nal with fur::kan mutation; Nalr, Kmr | This work |
| HKR.fur::kan-pJAK16 | HKR.fur::kan with pJAK16; Nalr, Kmr, Cmr | This work |
| HKR.fur::kan-pJAKfur | HKR.fur::kan with pJAK-fur; Nalr, Kmr, Cmr | |
| HKR(pJAK16) | HKR.Nal containing pJAK16; Nalr, Cmr | This work |
| HKR(pJAK-fur) | HKR.Nal containing pJAK-fur; fur overexpression strain; Nalr, Cmr | This work |
| HKR(pJAK16-JA03) | HKR.Nal containing pJAK-JA03; JA03 overexpression strain; Nalr, Cmr | This work |
| E. coli | ||
| TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (Smr) endA1 nupG phi80lacZΔM15 | Invitrogen |
| BL21(DE3) | fhuA2 (lon) ompT gal (λ DE3) (dcm) ΔhsdS λ DE3 = λ sBamHIo ΔEcoRI-B int::(lacI::PlacUV5::T7 gene1) i21 Δnin5 | Novagen |
| BL21(DE3)(pAaFur) | BL21(DE3) containing pAaFur | This work |
| SK140 | Top10 containing mobility plasmid pRK21761); Kmr | (Kachlany et al., 2000) |
| SK140(pJAK16) | SK140 containing pJAK16; Kmr, Cmr | This work |
| SK140(pJAK-fur) | SK140 containing pJAK16-fur; Kmr, Cmr | This work |
| SK140(pJAK16-JA03) | SK140 containing pJAK16-JA03; Kmr, Cmr | This work |
| H1717 | araD139 Δ(argF-lac) U169 rpsL150 relA1 flbB5301 deoC1 ptsF25 rbsR aroB fhuF::λ placMu53, Kmr, Smr, (fhuF-lacZ fusion, with fur) | (Hantke, 1987) |
| H1780 | araD139 Δ(argF-lac) U169 rpsL150 relA1 flbB5301 deoC1 ptsF25 rbsR fiu:: λplacMu53, (fiu-lacZ fusion, lacking fur) | (Hantke, 1987) |
| Plasmids | ||
| pGEM-T | TA cloning vector; Ampr | Promega |
| pGEMT-fur | pGEM-T containing HK1651R fur with engineered KpnI, BamHI sites; Ampr | This work |
| pGEMT-fur::kan | pGEM-T containing HK1651R fur::kan with engineered KpnI, BamHI sites; Ampr | This work |
| pGEMT-JA03 | pGEM-T containing HK1651R JA03 with engineered KpnI, BamHI sites; Ampr | This work |
| pGEMT-JA01pro | pGEM-T containing the HK1651R JA01 promoter; Ampr | This work |
| pGEMT-JA02pro | pGEM-T containing the HK1651R JA02 promoter; Ampr | This work |
| pGEMT-JA03pro | pGEM-T containing the HK1651R JA03 promoter; Ampr | This work |
| pGEMT-JA04pro | pGEM-T containing the HK1651R JA04 promoter; Ampr | This work |
| pET-30b | Novagen expression vector; Ampr | Novagen |
| pAaFur | pET-30b with the HK1651R fur gene cloned into the NcoI/NtoI sites; Ampr | This work |
| pRK21761 | RK2 IncP Kmr, Ampr, Tets, Lac+ (tetA::lacZ) oriT1 Mob+ | (Sia et al., 1996) |
| pMB78 | USS-containing cloning vector; suicide vector used for allelic exchange; Ampr | (Bhattacharjee et al., 2007) |
| pMfur | pMB78 containing HK1651R fur; Ampr | This work |
| pMfur.KanEZ | pMfur with insertion in fur gene; Ampr, Kmr | This work |
| pMJA03 | pMB78 containing HK1651R JA03; Ampr | This work |
| pJAK16 | IncQ expression plasmid; Cmr | (Thomson et al., 1999) |
| pJAK-fur | pJAK16 containing HK1651R fur; Cmr | This work |
| pJAK-JA03 | pJAK16 containing HK1651R JA03; Cmr | This work |
| Transposon | ||
| EZ::Tn5<KAN-2> | Tn5 transposon containing a Kmr cassette | Epicentre |
1
Amp; ampicillin; Cm, chloramphenicol; Km, kanamycin; Tet, tetracycline; r, resistant; s, sensitive; Sm, streptomycin.
2
ATCC, American Type Culture Collection.