Table 1.
Overview of constructed strains and plasmids used in this study.
| Strain | Genotype | Source |
|---|---|---|
| E. coli DH5α | φ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 56 hsdR17 (rk− mk+) supE44 thi-1 gyrA relA1 | Lab collection |
| B. subtilis 168 WT | trpC2 | DSM23778 |
| BS168 | B. subtilis 168 WT, pHT315 | This study |
| BS168 xyl | BS168::ΔaraR::PSGC14-SG46-araE, pJD18 | This study |
| BS168 amyQ | BS168, glms::P3P-amyQ::cm | This study |
| BS168 xyl amyQ | BS168 xyl, glms::P3P-amyQ::cm | This study |
| B. subtilis KO7 WT | ΔnprE, ΔaprE, Δepr, Δmpr, ΔnprB, Δvpr, Δbpr | Zeigler (2016) |
| KO7 | B. subtilis KO7 WT, pJD13 | This study |
| KO7 xyl | KO7::ΔaraR:: PSGC14-SG46-araE, pJD18 | This study |
| KO7 amyQ | KO7, glms::P3P-amyQ::cm | This study |
| KO7 xyl amyQ | KO7 xyl, glms::P3P-amyQ::cm | This study |
| B. subtilis KO7s | ΔnprE, ΔaprE, Δepr, Δmpr, ΔnprB, Δvpr, Δbpr, ΔsigF | Zeigler (2016) |
| KO7 | B. subtilis KO7s, pJD13 | This study |
| KO7s xyl | KO7s::ΔaraR:: PSGC14-SG46-araE, pJD18 | This study |
| KO7s amyQ | KO7s, glms::P3P-amyQ::cm | This study |
| KO7s xyl amyQ |
KO7s xyl, glms::P3P-amyQ::cm |
This study |
|
Plasmid |
Description |
Source |
| pJOE8999 | Plasmid containing Pmanp-cas9, pUC ori (E. coli), pE194ts ori (B. subtilis) and kanamycin resistance marker | (J, 2016) |
| pJD2 | pJOE8999 derivative containing araR homology regions | This study |
| pJD5 | pJOE8999- derivative containing ParaE homology regions | This study |
| for indel PSGC14-SG46 | Guiziou et al. (2016) | |
| pHT315 | Replicative plasmid containing pHT315 ori, ColE1 ori, ampicillin resistance, erythromycin resistance marker | Cavin et al. (1998) |
| pJD18 | pHT315 derivative containing PSCG7-SG46-araE and P43-xylA-xylB | This study |
| pProUSER13C1B | ProUSER2.0 plasmid containing P3P, glmS homology regions, and chloramphenicol resistance marker | Falkenberg et al. (2021) |
| pJD21 | pProUSER13C1B derivative for integrating P3P-amyQ downstream of glmS | This study |