Table 1.
Strains and plasmids used in this study. C. pa. = C. pasteurianum.
| Name | Designation | Properties | Source |
|---|---|---|---|
| C. pa. ATCC 6013 | CRG4080 | wild type, type strain | ATCC |
| C. pa. DSM 525 | CRG4091 | wild type, type strain | DSMZ |
| C. pa. DSM 525-H1 | CRG4111 | hypertransformable strain based on DSM 525 | This study |
| C. pa. DSM 525-H1ΔpyrE | CRG4273 | ACE pyrE truncation mutant | This study |
| C. pa. DSM 525-H1ΔpyrEΔspo0A | CRG5514 | ACE spo0A deletion mutant with ΔpyrE background | This study |
| C. pa. DSM 525-H1Δspo0A | CRG5516 | ACE spo0A deletion mutant with pyrE repaired | This study |
| C. pa. DSM 525-H1::spo0A* | CRG5518 | ACE spo0A complementation in pyrE locus | This study |
| C. pa. DSM 525-H1ΔpyrEΔrex | CRG5520 | ACE rex deletion mutant with ΔpyrE background | This study |
| C. pa. DSM 525-H1Δrex | CRG5522 | ACE rex deletion mutant with pyrE repaired | This study |
| C. pa. DSM 525-H1::rex* | CRG5524 | ACE rex complementation in pyrE locus | This study |
| C. pa. DSM 525-H1ΔpyrEΔhydA | CRG5526 | ACE hyd deletion mutant with ΔpyrE background | This study |
| C. pa. DSM 525-H1ΔhydA | CRG5528 | ACE hyd deletion mutant with pyrE repaired | This study |
| C. pa. DSM 525-H1::hydA* | CRG5530 | ACE hyd complementation in pyrE locus | This study |
| C. pa. DSM 525-H1ΔpyrEΔdhaBCE | CRG5532 | ACE dhaBCE deletion mutant with ΔpyrE background | This study |
| C. pa. DSM 525-H1ΔdhaBCE | CRG5534 | ACE dhaBCE deletion mutant with pyrE repaired | This study |
| C. pa. DSM 525-H1::dhaBCE* | CRG5536 | ACE dhaBCE complementation in pyrE locus | This study |
| E. coli Top10 x CR1 | CRG3131 | Strain harbouring plasmid CR1 With M.BepI methylase | This study |
| pMTL85151 | E. coli-Clostridium shuttle vector (pIM13, catP, ColE1 traJ, lacZα ORF/MCS, TCpa fdx) | Heap et al. (2009) | |
| pMTL-AMH101 | catP-pyrE module used for pMTL-KS15 | Liew et al. (2017) | |
| pMTL-KS01 | pMTL85151, 300-bp internal pyrE fragment, 1200-bp fragment immediately downstream of pyrE | This study | |
| pMTL-KS03 | pMTL85151, 300-bp internal pyrE fragment, 1200-bp fragment immediately downstream of pyrE, with TCtet gluRS | This study | |
| pMTL-KS04 | pMTL85151, 937-bp fragment immediately downstream of pyrE, 300-bp internal pyrE fragment, TCtet gluRS, TCpa fdx | This study | |
| pMTL-KS05 | pMTL85151, 548-bp internal pyrE fragment, 1200-bp fragment immediately downstream of pyrE | This study | |
| pMTL-KS08 | pMTL85151, 937-bp fragment immediately downstream of pyrE, 548-bp internal pyrE fragment, TCtet gluRS, TCpa fdx | This study | |
| pMTL-KS10 | pMTL-KS01 features, TCtet gluRS, TCpa fdx,AsiSI | This study | |
| pMTL-KS12 | pMTL85151, 548-bp internal pyrE fragment, 937-bp fragment immediately downstream of pyrE, TCtet gluRS, TCpa fdx,AsiSI | This study | |
| pMTL-KS15 | pMTL85151, pyrE (C. acetobutylicum), TCtet gluRS, TCpa fdx, 300-bp SHA pyrE, lacZα ORF/MCS | This study | |
| pMTL-KS16 | pMTL85151, pyrE (C. acetobutylicum), TCtet gluRS, TCpa fdx, AsiSI, 300-bp SHA pyrE, lacZα ORF/MCS | This study | |
| pMTL-AGH12 | 1748-bp fragment comprising pyrE (35–582 nt) and 1200-bp immediately downstream of pyrE cloned into SbfI/AscI recognition sites of pMTL-KS12 | This study | |
| pMTL-KS12::spo0A | 1095-bp fragment comprising the 267-bp sequence upstream of spo0A and the spo0A gene cloned into the NotI/NheI recognition sites of pMTL-KS12 | This study | |
| pMTL-KS12::rex | 816-bp fragment comprising the 183-bp sequence upstream of rex and the rex gene cloned into the NotI/NheI recognition sites of pMTL-KS12 | This study | |
| pMTL-KS12::hyd | 2118-bp fragment comprising the 393-bp sequence upstream of hyd and the hyd gene cloned into the NotI/NheI recognition sites of pMTL-KS12 | This study | |
| pMTL-KS12::dhaBCE | 2984-bp fragment comprising the 294-bp sequence upstream of dhaB and the dhaBCE genes cloned into the NotI/NheI recognition sites of pMTL-KS12 | This study | |
| pMTL-KS15::KO_spo0A | 1400-bp KO out cassette for spo0A | This study | |
| pMTL-KS15::KO_rex | 1364-bp KO out cassette for rex | This study | |
| pMTL-KS16::KO_hyd | 2002-bp KO out cassette for hyd | This study | |
| pMTL-KS15::KO_dhaBCE | 1602-bp KO out cassette for dhaBCE | This study |