TABLE 3.
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 |
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.