TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant genotype or phenotype | Source or reference |
|---|---|---|
| E. coli DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF) U169 endA1 recA1 hsdR1(rK− mK+) deoR thi-1 phoA supE44 λ−gyrA96 relA1 | Laboratory stock |
| B. subtilis | ||
| JH642 | trpC2 pheA1 | Laboratory stock |
| JCB50 | SPβc2 Δ2::Tn917::pSK10Δ6::Φ(spoVG′-lacZ) cat trpC2 pheA1 | 7 |
| JCB64 | Spβc2 Δ2::Tn917::pSK10Δ6::Φ(cotA′-lacZ) cat trpC2 pheA1 | 7 |
| KP66 | spoIIIG-lacZ spoIIIGΔ1 | 48 |
| SJB83 | SPβc2 Δ2::Tn917::pSK10Δ6::Φ(spoIIG′-lacZ) cat trpC2 pheA1 | 25 |
| SJB227 | Φ(sspB′-lacZ) cat trpC2 pheA1 | 7 |
| SJB253 | SPβc2 Δ2::Tn917::pSK10Δ6::Φ(spoIIA′-lacZ) cat trpC2 pheA1 | 25 |
| UOT-1101 | hisH metB Φ(spo0A′-lacZ) cat | 53 |
| AA001 | trpC2 pheA1 spoIIIG-lacZ spoIIIGΔ1 | KP66 DNA→JH642 |
| 501-75I | trpC2 pheA ???::neo | pUC18EAII-502→JH642 |
| 501-75I1 | trpC2 pheA ΔpdhA::neo pdhA | pUC18EAII-502→JH642 |
| 501-76 | trpC2 pheA1 ΔpdhB::neo | pDH87EB-502→JH642 |
| 501-760A | trpC2 pheA1 ΔpdhB::neo Φ(spo0A′-lacZ) cat | pDH87EB-502→UOT-1101 |
| 501-76IIA | trpC2 pheA1 ΔpdhB::neo SPβc2 Δ2::Tn917::pSK10Δ6::Φ(spoIIA′-lacZ) cat | pDH87EB-502→SJB253 |
| 501-76IIG | trpC2 pheA1 ΔpdhB::neo SPβc2 Δ2::Tn917::pSK10Δ6::Φ(spoIIG′-lacZ) cat | pDH87EB-502→SJB83 |
| 501-76IIIG | trpC2 pheA1 ΔpdhB::neo spoIIIG-lacZ spoIIIGΔ1 | pDH87EB-502→AA001 |
| 501-76VG | trpC2 pheA1 ΔpdhB::neo SPβc2 Δ2::Tn917::pSK10Δ6::Φ(spoVG′-lacZ) cat | pDH87EB-502→JCB50 |
| 501-76SSP | trpC2 pheA1 ΔpdhB::neo Φ(sspB′-lacZ) cat | pDH87EB-502→SJB227 |
| 50176COT | trpC2 pheAl ΔpdhB::neo SPβc2 Δ2::Tn917::pSK10Δ6::Φ(cotA′-lacZ) cat | pDH87EB-502→JCB64 |
| 501-77 | trpC2 pheA1 ΔpdhC::neo | pUC9EC-501→JH642 |
| 501-770A | trpC2 pheA1 ΔpdhC::neo Φ(spo0A′-lacZ) cat | pUC9EC-501→UOT-1101 |
| 501-77IIA | trpC2 pheA1 ΔpdhC::neo SPβc2Δ2::Tn917::pSK10Δ6::Φ(spoIIA′-lacZ) cat | pUC9EC-501→SJB253 |
| 501-77IIG | trpC2 pheA1 ΔpdhC::neo SPβc2 Δ2::Tn917::pSK10Δ6::Φ(spoIIG′-lacZ) cat | pUC9EC-501→SJB83 |
| 501-77IIIG | trpC2 pheA1 ΔpdhC::neo spoIIIG-lacZ spoIIIGΔ1 | pUC9EC-501→AA001 |
| 501-77VG | trpC2 pheA1 ΔpdhC::neo SPβc2 Δ2::Tn917::pSK10Δ6::Φ(spoVG′-lacZ) cat | pUC9EC-501→JCB50 |
| 501-77SSP | trpC2 pheA1 ΔpdhC::neo Φ(sspB′-lacZ) cat | pUC9EC-501→SJB227 |
| 501-77COP | trpC2 pheA1 ΔpdhC::neo SPβc2 Δ2::Tn917::pSK10Δ6::Φ(cotA′-lacZ) cat | pUC9EC-501→JCB64 |
| 501-78 | trpC2 pheA1 ΔpdhD::neo | pUC9ED-501→JH642 |
| Plasmids | ||
| pUC18 | E. coli cloning vector; Apr | Laboratory stock |
| pDH87 | B. subtilis integrational vector; Apr Camr | 18 |
| pUC18EA | B. subtilis HindIII-EcoRI fragment of pdhA in pUC18; Apr | This study |
| pUC18EA2 | B. subtilis pdhA in pUC18; Apr | This study |
| pDH87EB | B. subtilis HindIII-HindIII fragment of pdhAB in pDH87; Apr Camr | This study |
| pUC9EC | B. subtilis pKTH1877 fragment (containing pdhC) in pUC9; Apr | 16 |
| pUC9ED | B. subtilis pKTH1878 fragment (containing pdhD) in pUC9; Apr | 16 |
| pBEST501 | Neomycin gene donor vector; Apr Neor | 22 |
| pBEST502 | Neomycin gene donor vector; Apr Neor | 22 |
| pUC18EA-502 | Neomycin cassette at PstI site of pdhA of pUC18EA; Apr Neor | This study |
| pDH87EB-502 | Neomycin cassette at EcoRI site of pdhB of pDH87EB; Apr Neor | This study |
| pUC9EC-501 | Neomycin cassette at HpaI site of pdhC of pUC9EC; Apr Neor | This study |
| pUC9ED-501 | Neomycin cassette at SnaBI site of pdhC of pUC9ED; Apr Neor | This study |
| pMUTIN4 | Vector for systematic gene inactivation | 50 |
| pMUTIN4A | pMUTIN4 containing 800 bp of pdhA | This study |