Table 1.
Strains, plasmids, and oligonucleotides used
| Strain, plasmid, or primer | Genotype or sequencea | Reference or source | Plasmid constructed |
|---|---|---|---|
| E. coli strains | |||
| BW25113 | rrnBT14 ΔlacZWJ16hsdR514 ΔaraBADAH33 ΔrhaBADLD78 | 16 | |
| XL-1 Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lacIqZΔM15 Tn10 (Tetr)] | Stratagene | |
| JCL16 | BW25113/F′ [traD36 proAB+lacIqZΔM15 (Tetr)] | 2 | |
| JCL166 | JCL16 ΔldhA ΔadhE ΔfrdBC | 2 | |
| JCL299 | JCL16 ΔldhA ΔadhE ΔfrdBC Δpta | 2 | |
| Plasmids | |||
| pCS102 | PLlacO1::fdhCB p15A ori Kanr | This study | |
| pCS106 | PLlacO1::MCS Cola ori Kan | This study | |
| pCS127 | PLlacO1::terFS Cola ori Kanr | This study | |
| pCS128 | PLlacO1::terFJ Cola ori Kanr | This study | |
| pCS138 | PLlacO1:: fdhCB pSC101 ori Cmr | This study | |
| pEL11 | PLlacO1::atoBEC-adhE2CA-crtCA-hbdCA ColE1 ori Ampr | This study | |
| pEL12 | PLlacO1::atoBEC-crtCA-hbdCA ColE1 ori Ampr | This study | |
| pEL16 | PLlacO1::terTV Cola ori Kanr | This study | |
| pIM8 | PLlacO1::terTD Cola ori Kanr | This study | |
| pIM11 | PLlacO1::bcdCA-etfACA-etfBCA Cola ori Kanr | This study | |
| pHJ2 | PLlacO1::terFS(M11K/A397V) Cola ori Kanr | This study | |
| pHJ3 | PLlacO1::terFS(M11K/T325I) Cola ori Kanr | This study | |
| pHJ6 | PLlacO1::terFS(M11K) Cola ori Kanr | This study | |
| Primers | |||
| Fdhfacc65 | TCAGGTACCATGAAAATTGTGCTGGTGTTATATGATG | pCS138 | |
| Fdhrsal | TCAGTCGACTCACTTTTTATCATGTTTTCCGTAC | pCS138 | |
| Tdeterfacc65 | TCAGGTACCATGATTGTAAAACCAATGGTTAGGAACA | pIM8 | |
| Tdeterrsal | TCAGTCGACTAAATCCTGTCGAACCTTTCTACCTCG | pIM8 | |
| CacBcdfacc65 | TCAGGTACCATGGATTTTAATTTAACAAGAGAACAAGAAT | pIM11 | |
| CacEtfBrsal | TCAGTCGACTTAATTATTAGCAGCTTTAACTTGAGCTATTAATT | pIM11 | |
| Crtfsph | GGGAAAGCATGCAGGAGATATACCATGGAACTAAACAATGTCATCCTTGAAAAGG | pEL12 | |
| Crtfxba | GGGAAATCTAGAAGGAGATATACCATGGAACTAAACAATGTCATCCTTGAAAAGG | pEL11 | |
| CrtrSOE | CCTTTTTCATGGTATATCTCCTCTATCTATTTTTGAAGCCTTCAATTTTTCTTTTC | pEL11 | |
| HbdfSOE | AAATAGATAGAGGAGATATACCATGAAAAAGGTATGTGTTATAGGTGCAGG | pEL11 | |
| Hbdrxba | GGGAAATCTAGATTATTTTGAATAATCGTAGAAACCTTTTCCTGATT | pEL11 | |
| Tviterfacc65 | GGGAAAGGTACCATGAGTATGAAACCGATGCTGAGAA | pEL16 | |
| Tviterrsal | TCAGTCGACTTATATCCGGTCGAACCGGTCAATC | pEL16 | |
| Tviterrbam | GGGAAAGGATCCTTATATCCGGTCGAACCGGTCAATC | pEL16 | |
| Fsuterfacc65 | TCAGGTACCATGATTATCAAGCCGCTCATTCGT | pCS127 | |
| Fsuterrsal | ACGCAGTCGACTTAGATAGAGGTCAGGGTCTGAACATCT | pCS127 | |
| Fjoterfacc65 | TCAGGTACCATGATTATCGAACCGCGTATGCG | pCS128 | |
| Fjoterrsal | ACGCAGTCGACTTATTTGATAGATTCGATATTAACAACTTCGT | pCS128 |
a
In plasmid descriptions, subscripts indicate the source of the gene as follows: CA, Clostridium acetobutylicum; TD, Treponema denticola; TV, Treponema vincentii; FJ, Flavobacterium johnsoniae; FS, Fibrobacter succinogenes; CB, Candida boidinii. Primer sequences are shown 5′ → 3′.