Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant maker | Source |
|---|---|---|
| Escherichia coli | ||
| DH5α | F−, Φ80dlacZ∆M15, ∆(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17(rk−,mk+), phoA, supE44, l−, thi-1, gyrA96, relA1 | Takara Bio |
| S17-1 | thi, pro, hsdR, recA, chromosomal RP4, Tra+, Tmpr, Str/Spcr | Simon et al. 1983 |
| Ralstonia eutropha | ||
| H16 | Wild type | DSM 428 |
| NSDG-GGDB1 | H16 derivative; ∆phaC::phaCNSDG, ∆nagR, nagEGR, PA2858-glpFKEc-h16_A2858, ∆phaB1 | Zhang et al. 2019 |
| IF001 | NSDG-GG∆B1 derivative; ∆phaCNSDG::phaCAR | This study |
| IF002 | IF001 derivative; phaP1::ldhACd-hadACd | This study |
| IF005 | IF001 derivative; ∆h16_A0666 | This study |
| IF012 | IF001 derivative; ∆h16_B0460 | This study |
| IF013 | IF001 derivative; ∆h16_B1817 | This study |
| IF014 | IF001 derivative; ∆h16_A3091 | This study |
| IF015 | IF001 derivative; ∆h16_B0093-B0092-B0091 | This study |
| IF016 | IF001 derivative; ∆h16_A3091, ∆h16_B0093-B0092-B0091 | This study |
| IF017 | IF002 derivative; ∆h16_A3091 | This study |
| IF018 | IF002 derivative; ∆h16_B0093-B0092-B0091 | This study |
| IF019 | IF002 derivative; ∆h16_A3091, ∆h16_B0093-B0092-B0091 | This study |
| IF023 | IF001 derivative; ∆h16_A1681-A1682 | This study |
| Plasmids | ||
| pTTQ19 | pMB1 ori, Ampr, lacZα, lacIq, Ptac | Stark 1987 |
| pTTQ-ldhAhadACd | pTTQ19 derivative; ldhACd-hadACd (codon-optimized for E. coli) | Mizuno et al. 2018 |
| pBluescript II KS (+) | ColE1 ori, Ampr, lacZα, Plac | Agilent Technologies |
| pBSPRephaCARpct | pBluescript II KS (+) derivative; PphaC, phaCAR, pctMe | Matsumoto et al. 2018 |
| pK18mobsacB | pMB1 ori, RP4mob, Kmr, modified sacB, lacZα | Schäfer et al. 1994 |
| pK18ms-C61-3EDQKAB’ | pK18mobsacB derivative; ∆phaC::phaCEDQK | Laboratory stock |
| pK18ms-CARAB’ | pK18ms-C61-3EDQKAB’ derivative; ∆phaC::phaCAR | This study |
| pK18ms-P1udP1J | pK18mobsacB derivative; phaP1::phaJAc | Kawashima et al. 2015 |
| pK18ms-P1udP1-ldhACdhadACd | pK18ms-P1udP1J derivative; phaP1::ldhACd-hadACd | This study |
| pK18ms-A0666ud1000 | pK18ms derivative; h16_A0666 del | This study |
| pK18ms-B0460ud980 | pK18ms derivative; h16_B0460 del | This study |
| pK18ms-B1817ud980 | pK18ms derivative; h16_B1817 del | This study |
| pK18ms-A3091ud980 | pK18ms derivative; h16_A3091 del | This study |
| pK18ms-B0093-B0092-B0091ud980 | pK18ms derivative; h16_B0093-B0092-B0091 del | This study |
| pK18ms-A1681-A1682ud900 | pK18ms derivative; h16_A1681-A1682 del | This study |
| pBBR1MCS-2 | Broad host range plasmid; Kmr, mob, Plac, lacZα | Kovach et al. 1995 |
| pBPP | pBBR1MCS-2 derivative; PphaP1, TrrnB | Fukui et al. 2011 |
| pBPP-phaCAR | pBPP derivative; phaCAR | This study |
Ec, Escherichia coli; Ac, Aeromonas caviae; Cd, Clostridioides difficile; Me, Megasphaera elsdenii
del: Homologous regions for deletion of the target gene by homologous recombination