TABLE 2.
Bacterial strains and plasmids
| Strain or plasmid | Genotype or description | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | Lab stock | |
| BL21(DE3) pLysS | Novagen | |
| B. subtilis strains | ||
| JH642 | pheA1 trpC2 | J. A. Hoch |
| MH5562 | pheA1 trpC2 amy::pho-lacZ Cmr | 27 |
| MH7415 | pheA1 trpC2 scoC::(pJM2501) Cmr | This work |
| MH7416 | pheA1 trpC2 scoC::(pJM2501) Cms::pJL62 Spcr | This work |
| MH7400 | pheA1 trpC2 scoC::(pJM2501) Cms::pJL62 Spcramy::phoPR-lac Z Cmr | This work |
| MH7417 | pheA1 trpC2 scoC::(pJM2501) Cms::pJL62 Spcr pAT14Ω phoP cre1-lacZ Ermr | This work |
| MH6040 | pheA1 trpC2 pAT14Ω phoP cre1-lacZ Ermr | 32 |
| MH6024 | pheA1 trpC2 pAT3Ω phoP-lacZ Ermr | 32 |
| Plasmids | ||
| pJM2501 | A pUC19 plasmid carrying a 900-bp PvuII fragment containing the scoC gene with cat gene insertion mutation at PvuI | 28 |
| pJL62 | 1.1-Kb spectinomycin resistance cassette cloned into NcoI site of pJH101; Ampr Spcr Tetr | I. Smith |
| PCR2.1 | A linearized vector having single 3′ deoxythymidine (T) residues; Ampr Kanr | Invitrogen |
| pHT4-phoPR | pHT315::Sau3A1 fragment of B. subtilis chromosome containing 3′ region of mdh, phoP, and phoR and 5′ polA | 8; T. Masdek |
| pBK1 | 1.364-Kb SmaI-Pau I insert from pHT4-phoPR cloned into pUC18 | 26; this work |
| pBK6.1 | PCR-amplified scoC gene (636 bp) from JH642 chromosomal DNA ligated into PCR2.1 vector | This work |
| pBK6 | Vector for overexpression of ScoC; XhoI-BamHI fragment from pBK6.1 encoding scoC gene cloned into pET16b at the same restriction sites | This work |
| pAT3 | Full-length phoPR promoter from pES2 in pMUTIN2; bp −705 to +92 relative to phoP translation start site | 32 |
| pAT14 | phoP cre1 promoter from pAT12 in pMUTIN2; Ermr | 32 |
| pAT12 | phoP cre1 promoter in pCR2.1 | 32 |
| pSB5 | phoPR promoter in PCR2.1; Ampr Kanr; bp −301 to +92 relative to phoP translation start site | 27 |
| pSB40 | 396-bp BamHI/EcoRI fragment from pSB5 subcloned into pDH32; Ampr Cmr | 27 |