Table 1.
Strains and plasmids
| Strains and plasmids | Relevant characteristics | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| S17-1 | thi-1 proA hsdR17(rK− mK+) recA1; tra genes of plasmid RP4 integrated into the genome | 46 |
| XL10 Gold | endA1 glnV44 recA1 thi-1 gyrA96 relA1 lac Hte Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 tetR F′ [proAB lacIqZΔM15 Tn10(Tetr Amy Cmr)] | Stratagene |
| A. mimigardefordensis | ||
| DPN7 | Wild-type strain | 54 |
| M1 | DPN7 mutant (Δmdo) | This study |
| M2 | DPN7 mutant (ΔrecA) | This study |
| M3 | DPN7 mutant (Δmdo ΔrecA) | This study |
| M4 | DPN7 mutant (Δmdo recA::buk-ptb) | This study |
| M5 | DPN7 mutant (recA::buk-ptb) | This study |
| M6 | DPN7 mutant (Δmdo recA::buk-ptb ΔphaCAm) | This study |
| M7 | DPN7 mutant (Δmdo ΔrecA ΔphaCAm) | This study |
| SHX1 | DPN7 harboring pBBR1MCS5::BPEC | This study |
| SHX2 | DPN7 harboring pBBR1MCS5 | This study |
| SHX3 | DPN7 harboring pBBR1MCS5::phaEC | This study |
| SHX4 | DPN7 harboring pBBR1MCS5::buk-ptb | This study |
| SHX5 | DPN7 mutant (Δmdo recA::buk-ptb) harboring pBBR1MCS5::phaEC | This study |
| SHX6 | DPN7 mutant (Δmdo recA::buk-ptb) harboring pBBR1MCS5 | This study |
| SHX7 | DPN7 mutant (Δmdo) harboring pBBR1MCS5::BPEC | This study |
| SHX8 | DPN7 mutant (Δmdo) harboring pBBR1MCS5 | This study |
| SHX9 | DPN7 mutant (recA::buk-ptb) harboring pBBR1MCS5::phaEC | This study |
| SHX10 | DPN7 mutant (recA::buk-ptb) harboring pBBR1MCS5 | This study |
| SHX11 | DPN7 mutant (ΔrecA) harboring pBBR1MCS5::BPEC | This study |
| SHX12 | DPN7 mutant (ΔrecA) harboring pBBR1MCS5 | This study |
| SHX13 | DPN7 mutant (Δmdo ΔrecA) harboring pBBR1MCS5 | This study |
| SHX14 | DPN7 mutant (Δmdo ΔrecA ΔphaCAm) harboring pBBR1MCS5 | This study |
| SHX15 | DPN7 mutant (Δmdo recA::buk-ptb ΔphaCAm) harboring pBBR1MCS5 | This study |
| SHX16 | DPN7 mutant (Δmdo recA::buk-ptb ΔphaCAm) harboring pBBR1MCS5::phaEC_colinear | This study |
| SHX17 | DPN7 mutant (Δmdo recA::buk-ptb ΔphaCAm) harboring pBBR1MCS5::phaEC_antilinear | This study |
| SHX18 | DPN7 mutant (Δmdo recA::buk-ptb ΔphaCAm) harboring pBBR1MCS5::phaCAm_colinear | This study |
| SHX19 | DPN7 mutant (Δmdo recA::buk-ptb ΔphaCAm) harboring pBBR1MCS5::phaCAm_antilinear | This study |
| SHX20 | DPN7 mutant (Δmdo recA::buk-ptb ΔphaCAm) harboring pBBR1MCS5::phaC1Re_colinear | This study |
| SHX21 | DPN7 mutant (Δmdo recA::buk-ptb ΔphaCAm) harboring pBBR1MCS5::phaC1Re_antilinear | This study |
| SHX22 | DPN7 mutant (Δmdo recA::buk-ptb) harboring pBBR1MCS5::phaCAm_colinear | This study |
| Plasmids | ||
| pBBR1MCS5 | Broad-host-range vector, 4.9 kb, Gmr with lac promoter in front of MCS for white/blue selection | 23 |
| pBPP1 | Harbors the whole BPEC pathway genes | 29 |
| pBBR1MCS-5::phaEC_colinear | Harbors the operon of phaEC from Thiocapsa pfennigii colinear to lac promoter (Plac) at the locus of EcoRI site | This study |
| pBBR1MCS-5::buk-ptb | Harbors buk-ptb from Clostridium acetobutylicum colinear to Plac at the locus of BamHI site | This study |
| pBBR1MCS-5::BPEC | Harbors the phaEC operon at the locus of EcoRI site and the operon of buk-ptb at the locus of BamHI site together colinear to Plac | This study |
| pBBR1MCS-5::phaEC_antilinear | Harbors the operon of phaEC from Thiocapsa pfennigii antilinear to Plac at the locus of EcoRI site | This study |
| pBBR1MCS5::phaCAm_colinear | Harbors the operon of phaCAm from A. mimigardfordensis DPN7 colinear to Plac at the locus of HindIII site | This study |
| pBBR1MCS5::phaCAm_antilinear | Harbors the operon of phaCAm from A. mimigardfordensis DPN7 antilinear to Plac at the locus of HindIII site | This study |
| pBBR1MCS5::phaC1Re_colinear | Harbors the operon of phaC1 (phaC1Re) gene from Ralstonia eutropha H16 colinear to Plac at the locus of BamHI site | This study |
| pBBR1MCS5::phaC1Re_antilinear | Harbors the operon of phaC1 (phaC1Re) gene from Ralstonia eutropha H16 antilinear to Plac at the locus of BamHI site | This study |
| pJQ200mp18 | Gmr, suicide vector for gene deletion | 38 |
| pJQ200mp18Tc | Tcr, suicide vector for gene deletion | 37 |
| pJQ200mp18Tc::Δmdo | Tcr, suicide vector for mdo gene deletion | This study |
| pJQ200mp18::ΔrecA | Gmr, suicide vector for recA gene deletion | This study |
| pJQ200mp18::ΔrecA::Ωbuk-ptb | Gmr, suicide vector for buk-ptb operon sequence substitute for recA gene | This study |
| pJQ200mp18::ΔphaCAm | Gmr, suicide vector for phaC gene deletion | This study |