TABLE 1.
Bacterial strains, plasmids, and primers
| Strain, plasmid, or primer | Relevant characteristic(s) or sequence (5′-3′)a | Positionb | Reference or source |
|---|---|---|---|
| B. subtilis strains | |||
| 168 | trpC2 | 3 | |
| 168-PR | trpC2 phoRΔBalI::Tcr | 26 | |
| 168-SKFA | trpC2 skfA::pYBCOdd Emr (previously YBCOdd) | T. Tanaka, Takai University | |
| 168-SKFA-PR | trpC2 phoRΔBalI::TcrskfA::pYBCOdd Emr | This study | |
| SWV215 | trpC2 pheA1 spo0A::Kmr | 31 | |
| 168-SKFA-OA | trpC2 spo0A::KmrskfA::pYBCOdd Emr | This study | |
| 168-SKFA-OA-PR | trpC2 spo0A::KmrphoRΔBalI::TcrskfA::pYBCOdd Emr | This study | |
| 168-SKFB | trpC2 skfB::pYBCPdd Emr (previously YBCPdd) | T. Tanaka, Takai University | |
| Plasmids | |||
| pET-PhoP | pET2816 containing a 722-bp insert of phoP Apr (6.386 kb) | 25 | |
| pET-PhoR231 | pET2816 containing a 1,049-bp insert of phoR Apr (6.713 kb) | 25 | |
| Primers | |||
| skfA-FOR | AAGAATGGGAATCTGTGA | 213942-213960 | |
| skfA-REV | CTAATACGACTCACTATAGGGAGATAGCAGGATGCGGAAG | 214081-214066 | |
| skfB-FOR | CCAGAGTAGCGGTTCAT | 214181-214166 | |
| skfB-REV | CTAATACGACTCACTATAGGGAGTTACCTGAGCTGGCATAG | 214550-214532 | |
| skfH-FOR | CGTGACAAGTTGTGACGA | 219909-219892 | |
| skfH-REV | CTAATACGACTCACTATAGGGAGGTTGACTGAATGGCCAAG | 219611-219628 | |
| YbcO-FOR | TATAACATAATGGACCGTCT | 213645-213664 | |
| YbcO-REV | CACGTGTCATAGCAATACTC | 214035-214055 | |
| YhaX-FOR | TAACGATATTAGGGAGAATGGC | 1056018-1056039 | |
| YhaX-REV | GAAGCAGCGCTCCATCTATATT | 1056207-1056228 |
a
Sequences in boldface type are specific for the T7 promoter used to initiate in vitro transcription.
b
Positions of the primers specific for the B. subtilis 168 chromosome as indicated in the SubtiList database (http://genolist.pasteur.fr/SubtiList) (22).