Table 1.
Bacterial strains and gene expression plasmids
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| C. necator | ||
| H16 | Wild type | ATCC17699 |
| H16/ds | H16 derivative; ΔphaC1 | 17 |
| 005dZ126 | H16 derivative; ΔphaC1::phaC AcNSDG, ΔphaZ1, ΔphaZ2, ΔphaZ6 | 27 |
| CnC1A | 005dZ126, PphaC1RBS-phaJ4a | This study |
| CnTRPDA | 005dZ126, PtrpdRBS-phaJ4a | This study |
| CnTRPA | 005dZ126, PtrpRBS-phaJ4a | This study |
| CnUV5DA | 005dZ126, PlacUV5dRBS-phaJ4a | This study |
| CnUV5A | 005dZ126, PlacUV5RBS-phaJ4a | This study |
| CnTRCDA | 005dZ126, PtrcdRBS-phaJ4a | This study |
| CnC1DB | 005dZ126, PphaC1dRBS-phaJ4b | This study |
| CnC1B | 005dZ126, PphaC1RBS-phaJ4b | This study |
| CnTRPB | 005dZ126, PtrpRBS-phaJ4b | This study |
| CnUV5B | 005dZ126, PlacUV5RBS-phaJ4b | This study |
| CnTRCB | 005dZ126, PtrcRBS-phaJ4b | This study |
| E. coli | ||
| JM109 | recA1 endA1 gyrA96 thi hsdR17 supE44 relA1 Δ(lac-proAB)/F’ [traD36 proAB + lacI q lacZΔM15] | Takara |
| S17-1 | recA pro hsdR RP4-2-Tc::Mu-Km::Tn7 | ATCC47055 |
| Plasmids | ||
| pCUP3 | Stable plasmid vector in C. necator, Kmr | 17 |
| pCUP3-PphaC1RBS-phaC1 | PphaC1RBS-phaC1 expression cassette cloned into pCUP3 | This study |
| pCUP3-PphaC1dRBS-phaC1 | PphaC1dRBS-phaC1 expression cassette cloned into pCUP3 | This study |
| pCUP3-RBS-lacZ | RBS-lacZ cloned into pCUP3 | This study |
| pCUP3-PphaC1RBS-lacZ | PphaC1RBS-lacZ expression cassette cloned into pCUP3 | This study |
| pCUP3-PlacUV5RBS-lacZ | PlacUV5RBS-lacZ expression cassette cloned into pCUP3 | This study |
| pCUP3-PtrpRBS-lacZ | PtrpRBS-lacZ expression cassette cloned into pCUP3 | This study |
| pCUP3-PtrcRBS-lacZ | PtrcRBS-lacZ expression cassette cloned into pCUP3 | This study |
| pCUP3-dRBS-lacZ | dRBS-lacZ cloned into pCUP3 | This study |
| pCUP3-PphaC1dRBS-lacZ | PphaC1dRBS-lacZ expression cassette cloned into pCUP3 | This study |
| pCUP3-PlacUV5dRBS-lacZ | PlacUV5dRBS-lacZ expression cassette cloned into pCUP3 | This study |
| pCUP3-PtrpdRBS-lacZ | PtrpdRBS-lacZ expression cassette cloned into pCUP3 | This study |
| pCUP3-PtrcdRBS-lacZ | PtrcdRBS-lacZ expression cassette cloned into pCUP3 | This study |
aRBS, potential ribosome binding site of phaC1; dRBS, RBS disruptant