Table 1.
Bacillus strains and plasmids used in this study.
| Strain/plasmid | Genotype/description | References |
|---|---|---|
| BACILLUS STRAINS | ||
| B. subtilis 168 | Tryptophan auxotrophic (trpC2) | Laboratory stock |
| BS-C100 | B. subtilis 168 containing plasmid pHCas9 | So et al., 2017 |
| BS-D119a | BS-C100 containing plasmid pSC1 | This study |
| BS-D119b | BS-C100 ΔaprE containing plasmid pG2-aE | This study |
| BS-D119c | BS-C100 ΔaprE containing plasmid pG-aE | This study |
| BS-D119 | BS-C100 ΔaprE | This study |
| BS-D120 | BS-C100 ΔaprE ΔnprE | This study |
| BS-D121 | BS-C100 ΔaprE ΔnprE Δepr | This study |
| BS-D122 | BS-C100 ΔaprE ΔnprE Δepr Δbpr | This study |
| BS-D123 | BS-C100 ΔaprE ΔnprE Δepr Δbpr Δmpr | This study |
| BS-D124 | BS-C100 ΔaprE ΔnprE Δepr Δbpr Δmpr ΔnprB | This study |
| BS-D125 | BS-C100 ΔaprE ΔnprE ΔeprΔbpr Δmpr ΔnprB Δvpr | This study |
| BS-D126 | BS-C100 ΔaprEΔnprEΔepr Δbpr Δmpr ΔnprB Δvpr ΔwprA | This study |
| PLASMIDS | ||
| pAD123 | E. coli-Bacillus shuttle vector | Bacillus Genetic Stock Center |
| pAgR | Plasmid pAD123 derivative containing synthetic sgRNA module | So et al., 2017 |
| pSC1 | Plasmid pAD123 derivative containing sgRNA module with 20 bp self-targeting gRNA under the control of sporulation-specific promoter PspoIV A | This study |
| pG2 | Plasmid pSC1 containing synthetic sgRNA module under the control of the constitute promoter (Para) | This study |
| pG2-aE | Plasmid pG2 containing aprE-targeting 20 bp gRNA and 1 kb Donor DNA | This study |
| pG-aE | Plasmid pG2-aE without sporulation-specific promoter PspoIV A and 20 bp self-targeting gRNA | This study |
| pG2-nE | Plasmid pG2 containing nprE-targeting 20 bp gRNA and 1 kb Donor DNA | This study |
| pG2-er | Plasmid pG2 containing epr-targeting 20 bp gRNA and 1 kb Donor DNA | This study |
| pG2-br | Plasmid pG2 containing bpr-targeting 20 bp gRNA and 1 kb Donor DNA | This study |
| pG2-mr | Plasmid pG2 containing mpr-targeting 20 bp gRNA and 1 kb Donor DNA | This study |
| pG2-nB | Plasmid pG2 containing nprB-targeting 20 bp gRNA and 1 kb Donor DNA | This study |
| pG2-vr | Plasmid pG2 containing vpr-targeting 20 bp gRNA and 1 kb Donor DNA | This study |
| pG2-wA | Plasmid pG2 containing wprA-targeting 20 bp gRNA and 1 kb Donor DNA | This study |