TABLE 1.
B. subtilis strains and plasmids used in this study
| Strain or plasmid | Genotype or descriptiona | Reference or source |
|---|---|---|
| Strains | ||
| CU741 | trpC2 leuC7 | S. A. Zhaler |
| OAM145 | trpC2 leuC7 amyE::aprE-lacZ(−412 [SG35.18], Cmr) | 10 |
| OAM146 | trpC2 leuC7 amyE::aprE-lacZ(−340 [SG35.21], Cmr) | 10 |
| OAM147 | trpC2 leuC7 amyE::aprE-lacZ(−299, Cmr) | 10 |
| OAM218 | trpC2 leuC7 amyE::aprE-lacZ(−267, Cmr) | 10 |
| TT715 | trpC2 leuC7 aprE-lacZ (Cmr) | 7 |
| MU38 | trpC2 leuC7 amyE::degR-lacZ (Cmr) mecA::KmrdegU::Spr | 12 |
| TU38 | trpC2 leuC7 aprE-lacZ (Cmr) degU::Spr | MU38→TT715 |
| OAM157 | trpC2 leuC7 amyE::aprE-lacZ(−412 [SG35.18], Cmr) scoC::Emr (lacZ::Tcr) | 10 |
| OAM221 | trpC2 leuC7 amyE::aprE-lacZ(−412 [SG35.18], Cmr) sinR::Pmr | 10 |
| OAM169 | trpC2 leuC7 amyE::aprE-lacZ(−412 [SG35.18], Cmr) spoOA::SprabrB::Kmr | 10 |
| TSU2 | trpC2 leuC7 amyE::scoC-lacZ (Cmr) | pSCO1→CU741 |
| KAW1 | trpC2 leuC7 amyE::scoC-lacZ (Cmr) spoOA::Spr | OAM169→TSU2 |
| Plasmids | ||
| pDG148 | Multicopy B. subtilis and Escherichia coli plasmid; Apr, Kmr | 21 |
| pSEN24 | pDG148 carrying senS | This study |
| pIS284 | E. coli plasmid for insertion of lacZ fusions into the B. subtilis amyE locus | I. Smith |
| pSCO1b | pIS284 carrying scoC-lacZ at the amyE locus | This study |
The numbers in parentheses and the letters in brackets indicate the deletion end points upstream of the transcription start point of aprE and promoter constructs, respectively. Emr, erythromycin resistance; Tcr, tetracycline resistance; Pmr, phleomycin resistance; Cmr, chloramphenicol resistance; Kmr, kanamycin resistance; Spr, spectinomycin resistance; Apr, ampicillin resistance. The concentrations of the antibiotics added to the medium were 1 μg/ml for erythromycin, 5 μg/ml for chloramphenicol and phleomycin, 15 μg/ml for neomycin, 10 μg/ml for tetracycline, and 100 μg/ml for spectinomycin.
Plasmid pSCO1 was constructed as follows. A 561-bp PCR fragment spanning nt 76 in the yhaH gene located immediately upstream of scoC to nt 102 in scoC was prepared with primers YhaHF (5′-AGTTGAATTCGCACCTTCCTCAGGAAAGC-3′) and ScoCR (5′-AGTTGGATCCTTCTCGATCGATTTCC-3′) and CU741 DNA as a template, digested with EcoRI and BamHI (sites are underlined), and inserted into the EcoRI and BamHI sites of pIS284.