TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference(s) |
|---|---|---|
| A. caviae | ||
| FA440 | Wild type | FERM P-3432; 8, 26 |
| AC004 | PHA-negative mutant of FA440 | This study |
| E. coli | ||
| DH5α | deoR endA1 gyrA96 hsdR17(rK− mK+) recA1 relA1 supE44 thi-1 Δ(lacZYA-argFV169) φ80 ΔlacZΔM15 F− r− | Clontech |
| S17-1 | recA and tra genes of plasmid RP4 integrated into chromosome; auxotrophic for proline and thiamine | 27 |
| BL21(DE3) | F−ompT hsdSB(rB− mB−) gal dcm (DE3) | Novagen |
| pJRD215 | Cosmid; KmrSmr RSF1010 replicon; Mob+ | 5 |
| pJRDEE32 | 3.2-kbp insert (EcoRI fragment containing phaCAc, ORF1, and ORF3 [phaJAc]) in pJRD215 | 11 |
| pJRDEE32d1 | 2.8-kbp insert (EcoRI fragment containing phaCAc and ORF3 [phaJAc]) in pJRD215 | 11 |
| pJRDEE32d3 | 2.8-kbp insert (EcoRI fragment containing phaCAc and ORF1) in pJRD215 | 11 |
| pJRDEE32d13 | 2.4-kbp insert (EcoRI fragment containing phaCAc) in pJRD215 | 11 |
| pJRDG13 | 1.4-kbp insert (EcoRI fragment containing ORF1 and ORF3 [phaJAc]) in pJRD215 | This study |
| pEE32 | 3.2-kbp insert (EcoRI fragment containing phaCAc, ORF1, and ORF3 [phaJAc]) in pUC18 | 11 |
| pET-3a | Apr; contains a T7 promoter with start codon and designed ribosome binding site | Novagen |
| pETNB3 | 0.4-kbp insert (PCR-amplified NdeI-BamHI fragment of ORF3 [phaJAc]) in pET-3a | This study |