TABLE 1.
B. subtilis strains and plasmids used in this study
| Strain or plasmid | Genotype or structurec | Source, reference, or construction |
|---|---|---|
| Strains | ||
| BGC5 | trpC2 lys-3 bglS::cat′-erm | pCm::Era→BGW7 (27) |
| BGC9 | GM1133 with ΔlicT-bglS::erm | pSK3→GM1133 |
| BGC12 | GM1210 with amyE::(bglP′-lacZ cat) | pSL3→GM1210 |
| BGC13 | BGC12 with bglPH′-aphA3 phl | pCL18→BGC12 |
| BGC14 | BGC13 with ΔptsGHI::tet | GM1220→BGC13 |
| BGC14-pia1 | BGC14 with licT(Pia) (Phe211Val) | This study |
| BGC14-pia2 | BGC14 with licT(Pia) (Phe211Val) | This study |
| BGC14-pia3 | BGC14 with licT(Pia) (Arg124Trp) | This study |
| BGC14-pia4 | BGC14 with licT(Pia) (Asp 99Asn) | This study |
| BGC14-pia5 | BGC14 with licT(Pia) (Phe211Cys) | This study |
| BGC14-pia6 | BGC14 with licT(Pia) (Arg124Leu) | This study |
| BGC14-pia7 | BGC14 with licT(Pia) (Arg124Trp) | This study |
| BGC14-pia8 | BGC14 with licT(Pia) (Arg124Leu) | This study |
| BGC14-pia9 | BGC14 with licT(Pia) (Arg124Trp) | This study |
| BGC14-piaXb | BGC14-piaX with amyE::(bglP′-lacZ Δcat) | This study |
| BGC50 | BGC12 with amyE::(bglP′-lacZ Δcat) | This study |
| BGC51 | BGC50 with ΔlicT-bglS::erm | BGC9→BGC50 |
| BGC54-piaX | BGC41-piaX with bglS::cat | BGW7→BGC41-piaX |
| BGC72 | BGC50 with bglS::cat | BGW7→BGC51 |
| BGC72-piaX | BGC50 with licT(PiaX) bglS::cat | BGC54-piaX→BGC51 |
| BGC88 | BGC72 with ccpA::Tn917spc | GM1225→BGC72 |
| BGC88-piaX | BGC72-piaX with ccpA::Tn917spc | GM1225→BGC72-piaX |
| BGW7 | trpC2 lys-3 bglS::cat | 25 |
| GM1133 | sacXYΔ3 sacBΔ23 sacTΔ4 amyE::(sacB′-′lacZ phl) bglPΔ3 | 16 |
| GM1210 | ΔlacA sacXYΔ3 sacTΔ23 sacTΔ4 | Laboratory stock |
| GM1220 | GM1210 with ΔptsGHI::tet | Laboratory stock |
| GM1225 | trpC2 pheA1 Δ(bgaX) amyE::(gntRK′-lacZ phl) ccpA::Tn917 spc | 9 |
| Plasmids | ||
| pBGW3 | Ampr Tetr pBR322 derivative containing licT-bglS | 25 |
| pCL18 | Spcr containing the bglPH′-aphA3 phl construction | This study |
| pCL46 | E. coli: Ampr; B. subtilis: Ermr with the temperature-sensitive origin of pE194, containing the amyE::(bglP′-lacZ) fusion of pSL3, but cat is deleted | This study |
| pIC22 | pUC18 derivative with Phlr of pUB110 | 27 |
| pIC239 | Ampr Camr promoterless aphA3 | M. Steinmetz |
| pIC254 | E. coli: Ampr; B. subtilis: Ermr with the temperature-sensitive origin of pE194 containing an 11-kb B. subtilis sequence including bglPH | 26 |
| pIC408 | Spcr ori pUC multiple cloning site of pBluescript II SK(+) | M. Steinmetz |
| pSK3 | Ampr Tetr, pBGW3 with licT-bglS::erm | 15 |
| pSL3 | AmpramyE::(bglP′-lacZ) cat | 15 |
a
Arrows indicate construction by transformation.
b
“piaX” stands for pia1, pia4, pia5, pia6, and pia9.
c
Ampr, Tetr, Spcr, and Ermr, resistance against ampicillin, tetracycline, spectinomycin, and erythromycin, respectively.