TABLE 1.
Bacterial strains and plasmids used
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Strains | ||
| B. subtilis | ||
| 1A594 | trpC2 hemG321 | BGSCa |
| 168 | Wild type | BGSC |
| 3G18Δ2 | trpC2 met ade ΔhemY::ble | 9 |
| 1012 | leuA8 metB5 trpC2 hsrM1 | 35 |
| AM01 | 1012 with amyE::cat | 28 |
| AM7 | Promoterless bgaB inserted at amyE | 28 |
| AM10 | lepA::bgaB transcriptional fusion at amyE | 28 |
| ARB1 | HZ04 with resDE mutant from MH5081 | This work |
| ARB2 | HZ04 with fnr mutant from THB2 | This work |
| ARB3 | HZ04 with ywiD mutant from THB99 | This work |
| ARB4 | AM10 with resDE mutant from MH5081 | This work |
| ARB5 | AM10 with fnr mutant from THB2 | This work |
| ARB6 | AM10 with ywiD mutant from THB99 | This work |
| ARB7 | AM7 with resDE mutant from MH5081 | This work |
| ARB8 | AM7 with fnr mutant from THB2 | This work |
| ARB9 | AM7 with ywiD mutant from THB99 | This work |
| ARB10 | HZ03 with hemY mutation from ARB11, hemN hemZ hemY triple mutant | This work |
| ARB11 | 1012 carry hemY mutant introduced by using pMUTIN-hemY | This work |
| HZ01 | 1012 with 501-bp in-frame deletion in hemN, previously described as hemNΔ | 17 |
| HZ02 | 1012 with replacement of a part of hemZ by a cat cassette | This work |
| HZ03 | HZ01 with part of hemZ replaced by a cat cassette, hemN hemZ double mutant | This work |
| HZ04 | 1012 with hemZ::bgaB transcriptional fusion inserted at amyE | This work |
| JH642 | pheA1 trpC2 | BGSC |
| MH5081 | trpC2 pheA1 ΔresDE::tet | 39 |
| THB2 | trpC2 pheA1 fnr::spec | 14 |
| THB99 | trpC2 pheA1 ywiD::spec | 27 |
| E. coli | ||
| DH10B | Host in cloning experiments | Gibco BRL |
| DH5α | hsdR recA1 lacZYA f80 lacZDM15 gyrA96 | 36 |
| S. typhimurium TE3006 | env-53 hemN704::Mud-J(b) hemF707::Tn10d-Tet | 47 |
| Plasmids | ||
| pALF01 | pBluescript SK+II with 783-bp PCR product carrying the complete former yhaW and the 5′ end of former yhaV | This work |
| pBgaB | Promoter-test vector containing the bgaB gene | 28 |
| pBluehemZ | pBluescript SK+II with 1,706-bp PCR product carrying hemZ and its promoter | This work |
| pBlueSalISmaIΔ | pBluescript SK+II with deletion of the polylinker region between and including SalI and SmaI | 17 |
| pBluescript SK+II | Cloning vector | Stratagene |
| pBluescript KS+II | Cloning vector | Stratagene |
| pGH02 | 1,256-bp ClaI fragment in pBluescript KS+ containing lepA and its 5′ region | 17 |
| phemNΔ | PCR product of the mutated hemN gene from B. subtilis hemNΔ in pBluescript KS+II | This work |
| phemZ-PEX | pBluescript SK+ carrying 677-bp PCR product of former yhaW and its 5′ region | This work |
| phemZ1RPT3 | pBluescript SK+II carrying 500-bp PCR product of former yhaW | This work |
| phemZ2RPT3 | pBluescript SK+II carrying 877-bp PCR product of former yhaV | This work |
| phemZΔ01 | Fusion of a 149-bp PCR product carrying the 5′ region of former yhaV with a 153-bp fragment of the 3′ end of hemZ in pBluescript SK+II | This work |
| phemZΔ02 | 316-bp BamHI fragment from phemZΔ01 in pBlueSalISmaIΔ01 | This work |
| phemZ::cat | cat cassette integrated in EcoRV site of the phemZΔ02 insert | This work |
| pBluehemN | B. subtilis hemN in pBluescript SK+II | 13 |
| pBluehemZ::cat | PCR product of the tagged hemZ gene from B. subtilis hemZ::cat in pBluescript KS+II | This work |
| pHZ01 | 175-bp hemZ promoter fused to bgaB in pBgaB | This work |
| pMUTIN-hemY | pMUTIN4 with 324-bp fragment of hemY | This work |
| pMUTIN4 | Integration vector | 44 |
| pOKBShemN | 1,302-bp fragment containing the B. subtilis hemN gene in pOK12 | 13 |
| pSKCAT | cat cassette cloned into EcoRV site of pBluescript SK+II | This work |
a
BGSC, Bacillus Genetic Stock Center.