TABLE 2.
Strains and plasmids used in the study
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | λ− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK−) supE44 thi-1 gyrA relA1 | Stratagene (La Jolla, CA) |
| T. kodakarensis | ||
| KOD1 | Wild type | 7, 8 |
| KU216 | KOD1 ΔpyrF | 20 |
| KC01 | KU216 Pcsg-chiA | This study |
| KC04 | KU216 Pcsg-chiA Pgdh-dppABCDF-glmDA EF-1α gene promoter-dac | This study |
| KU216Δt | KU216 Δtgr | This study |
| KC04Δt | KC04 Δtgr | This study |
| KC04ΔtM1 | KC04Δt, after adaptive engineering | This study |
| Plasmids | ||
| pUC118 | General cloning vector, Ampr | TaKaRa Bio (Otsu, Japan) |
| pUD3 | pUC118 derivative; pyrF marker cassette | 27 |
| pUD3(ΔNdeI) | pUD3 derivative; NdeI site was deleted | This study |
| pUD3-Pcsg-chiA | pUD3(ΔNdeI) derivative; Pcsg-chiA pyrF | This study |
| pUD3-dac-Pef-1α-Pcsg-chiA | pUD3(ΔNdeI) derivative; Pcsg-chiA EF-1α gene promoter-dac pyrF | This study |
| pUD3-Pgdh-dppA | pUD3(ΔNdeI) derivative; Pgdh-dppA pyrF | This study |
| pUD3_1769D | pUD3 derivative; for tgr disruption | This study |
| pLC70 | pLC64 derivative; hmg trpE | 17 |
| pLC70M | pLC70 derivative; hmg | This study |
| pLC-tgr | pLC70 derivative; hmg tgr | This study |