Table 1.
Strains and plasmids used in this work
| Strain or plasmid | Genotype/Description | Origin/Reference |
|---|---|---|
| E. coli | ||
| TOP10 | F- mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 nupG recA1 araD139 Δ(ara-leu)7697 galE15 galK16 rpsL(StrR) endA1 λ−; StrR | Invitrogen |
| B. subtilis | ||
| 1A1 | trpC2 | BGSC* |
| LMD100 | trpC2 spoVDΩpLEB2; CmR | This work |
| LMD101 | trpC2 ΔspoVD | This work |
| LMD104 | trpC2 ΔspoVD amyE::PspoVD-spoVD-mCherry; SpR | This work |
| LMD115 | trpC2 ΔspoVD amyE:: PspoVD -spoVD(Ser294Ala)-mCherry; SpR | This work |
| Plasmids | ||
| pJM103-I-SceI | Suicide integration vector pJM103 (Perego, 1993) with I-SceI restriction site; ApR, CmR | Perego (1993) |
| pBKJ223 | I-SceI expression vector; ApR, TcR | Janes & Stibitz (2006) |
| pDG1730 | amyE integration vector; ApR, SpR, EryR | Guerout-Fleury et al. (1996) |
| pKS-mCherry-E-T3 | E. coli vector for generating gene fusions with mCherry; ApR | N. Ausmees |
| pLEB1 | 289 bp region upstream of spoVD cloned into pJM103-I-SceI; ApR, CmR | This work |
| pLEB2 | 336 bp region downstream of spoVD cloned into pLEB1; ApR, CmR | This work |
| pLEB5 | pKS-mCherry-E-T3 with a 2.0-kb fragment containing PspoVD –spoVD | This work |
| pLEB6 | pDG1730 with a 2.8-kb fragment containing PspoVD -spoVD-mCherry gene fusion; ApR, SpR, EmR | This work |
| pLEB19 | pDG1730 with a 2.8-kb fragment containing PspoVD -spoVD(Ser294Ala)-mCherry; AmpR, SpR, EmR | This work |
*
Bacillus Genetic Stock Center, Columbus, OH.
Ap, ampicillin; Cm, chloramphenicol; Em, erythromycin; Sp, spectomycin, Str, streptomycin; Tc, tetracycline.