Table 1 Strains and plasmids used in this study
Strains and plasmids | Relevant characteristics | Source or reference |
|---|---|---|
Strains | ||
C. saccharoperbutylacetonicum N1-4 (HMT) | N1-4, DSM 14923 (=ATCC 27021), WT strain | DSMZ |
C. saccharoperbutylacetonicum ΔR1 | Derived from N1-4, ΔCspa_c00280 | This study |
C. saccharoperbutylacetonicum ΔR2 | Derived from N1-4, ΔCspa_c21720 | This study |
C. saccharoperbutylacetonicum ΔR3 | Derived from N1-4, ΔCspa_c27220 | This study |
C. saccharoperbutylacetonicum N1-4-dcas9-R4 | N1-4 with the expression of pYW-19d-qssR4 | This study |
C. saccharoperbutylacetonicum ΔR5 | Derived from N1-4, ΔCspa_c56960 | This study |
C. saccharoperbutylacetonicum ΔR1-R1 | ΔR1 strain with the expression of pMTL-qssR1 | This study |
C. saccharoperbutylacetonicum ΔR2-R2 | ΔR2 strain with the expression of pMTL-qssR2 | This study |
C. saccharoperbutylacetonicum ΔR3-R3 | ΔR3 strain with the expression of pMTL-qssR3 | This study |
C. saccharoperbutylacetonicum N1-4-R4 | N1-4 strain with the expression of pMTL-qssR4 | This study |
C. saccharoperbutylacetonicum ΔR5-R5 | ΔR5 strain with the expression of pMTL-qssR5 | This study |
C. saccharoperbutylacetonicum N1-4 (pMTL82151) | N1-4 strain with the expression of pMTL82151 | This study |
Clostridium tyrobutyricum ATCC 25755 | ATCC 25755 (= KCTC 5387) | ATCC |
E. coli DH5α | F−, φ80dlacZΔM1, Δ(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17(r −k , m +k ), phoA, supE44, λ−thi-1, gyrA96, relA1 | NEB |
Plasmids | ||
pYW34 | CAK ori, Ampr, Ermr, Plac::Cas9, gRNA | [24] |
pMTL82151 | pBP1 ori, catP, ColE1, tra | [47] |
pYW19d-BseRI | CAK ori, Ampr, Ermr, Pthl::dCas9, gRNA | [48] |
pYW34-∆qssR1 | Derived from pYW34, J23119::20-nt gRNA targeting Cspa_c00280, homology arms | This study |
pYW34-∆qssR2 | Derived from pYW34, J23119::20-nt gRNA targeting Cspa_c21720, homology arms | This study |
pYW34-∆qssR3 | Derived from pYW34, J23119::20-nt gRNA targeting Cspa_c27220, homology arms | This study |
pYW34-∆qssR5 | Derived from pYW34, J23119::20-nt gRNA targeting Cspa_c56960, homology arms | This study |
pMTL-qssR1 | Derived from pMTL82151, with the expression of Cspa_c00280 under the control of cat1 promoter from C. tyrobutyricum ATCC 25755 | This study |
pMTL-qssR2 | Derived from pMTL82151, with the expression of Cspa_c21720 under the control of cat1 promoter from C. tyrobutyricum ATCC 25755 | This study |
pMTL-qssR3 | Derived from pMTL82151, with the expression of Cspa_c27220 under the control of cat1 promoter from C. tyrobutyricum ATCC 25755 | This study |
pMTL-qssR4 | Derived from pMTL82151, with the expression of Cspa_c29260 under the control of cat1 promoter from C. tyrobutyricum ATCC 25755 | This study |
pMTL-qssR5 | Derived from pMTL82151, with the expression of Cspa_c56960 under the control of cat1 promoter from C. tyrobutyricum ATCC 25755 | This study |
pYW19d-qssR4 | Derived from pYW19d-BseRI, with J23119::20-nt gRNA targeting Cspa_c29260 | This study |