TABLE 3.
Strains and plasmids used in this study
| Strain or plasmid | Characteristicsa | Source or reference |
|---|---|---|
| Strains | ||
| E. coli DH5α | F− endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG purB20 ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 hsdR17(rK− mK+) λ− | NEB |
| C. beijerinckii NCIMB 8052 | ATCC 51743 | ATCC |
| C. saccharoperbutylacetonicum | ||
| N1-4 | ATCC 13564, wild-type strain | ATCC |
| deltpta | N1-4 Δpta | This work |
| deltbuk | N1-4 Δbuk | This work |
| deltptabuk | N1-4 Δpta Δbuk | This work |
| Plasmids | ||
| pYW34 | CAK1 ori Ampr Ermr Plac::Cas9 gRNA | 26 |
| pYW34-pta | pYW34 PsRNA::20-nt gRNA targeting pta | This work |
| pYW34-deltpta | pYW34-pta::homology arm sequences | This work |
| pYW34-buk | pYW34 PsRNA::20-nt gRNA targeting buk | This work |
| pYW34-deltbuk | pYW34-buk::homology arm sequences | This work |
| pSHW1-pta | pYW34 Pvegb::20-nt gRNA targeting pta | This work |
| pSHW2-pta | pYW34 Pvegc::20-nt gRNA targeting pta | This work |
| pSHW3-pta | pYW34 PJ23119::20-nt gRNA targeting pta | This work |
| pSHW1-deltpta | pSHW1-pta::homologous arm sequences | This work |
| pSHW2-deltpta | pSHW2-pta::homologous arm sequences | This work |
| pSHW3-deltpta | pSHW3-pta::homologous arm sequences | This work |
a
PsRNA, small RNA promoter from C. beijerinckii; Pvegb, veg promoter from B. subtilis; Pvegc, hypothetical veg gene promoter from C. saccharoperbutylacetonicum; PJ23119, J23119 promoter from E. coli.