TABLE 1

Bacterial strains and plasmids used in this study

Bacterial strain or plasmidRelevant characteristicsaSource or reference
Strains
    E. coli strains
        TurbolacIq endA1 Δ(hsdS-mcrB)5NEB
        10-beta competent strainrecA1 endA1 Δ(mrr-hsdRMS-mcrBC)NEB
        BL21(pAN3)pAN3 Kmr ϕ3T I35
    C. acetobutylicum strains
        ATCC 824 (WT)Wild-type strainATCC
        824(p94MCS) (empty plasmid strain)p94MCS; MLSr35
        824(p94_solB) (solB overexpression strain)p94_solB; MLSrThis study
        824(pKO_mazF::solB)pKO_mazF::solB; ThrThis study
        824 ΔsolBsolB deletion; ThrThis study
Plasmids
    p94MCSAmpr MLSr; ptb promoter; MCS35
    p94MCS_solBAmpr MLSr; ptb promoter; solBThis study
    pKO_mazFThr-FRT; repL ori; bgaR and Pbgal upstream of mazF35
    pKO_mazF::solBThr-FRT; repL ori; bgaR and Pbgal upstream of mazF; Thr cassette flanked by ∼900-bp upstream and downstream solB regions of homologyThis study
    pAN3Kmr; ϕ3T I35
  • a lacIq, lac repressor gene; endA1, nonspecific endonuclease gene deleted; Δ(hsdS-mcrB)5, host-specific restriction and methylcytosine-specific restriction genes deleted; recA1, homologous recombination gene deleted; Δ(mrr-hsdRMS-mcrBC), host-specific restriction and methylcytosine-specific restriction genes deleted; Kmr, kanamycin resistance; ϕ3T I, Bacillus subtilis phage ϕ3T I methyltransferase gene; MLSr, macrolide-lincosamide-streptogramin B resistance; Thr, thiamphenicol resistance; Ampr, ampicillin resistance; ptb, phosphotransbutyrylase gene; MCS, multiple cloning site; Thr-FRT, FRT-flanked thiamphenicol resistance gene; repL, pIM13 Gram-positive origin of replication; ori, ColE1 origin of replication; bgaR, beta-galactosidase regulator; Pbgal, promoter of beta-galactosidase; mazF, codon-optimized E. coli MazF toxin gene; pUC ori, pUC Gram-negative origin of replication.