TABLE 1.
Bacterial strains and plasmids used
| Bacterial strain or plasmid | Relevant genotype and/or properties | Source or reference |
|---|---|---|
| Strains | ||
| Sphingomonas sp. strain CHY-1 | Wild type; Phe+ Chry+ | This study |
| Sphingomonas sp. strain M10-1 | Mutant of CHY-1 carrying a phnA1a::Gmr deletion-insertion; Phe− Chry− | This study |
| Pseudomonas putida KT2442 | Rifr | 10 |
| Escherichia coli DH5α | F−endA1 hsdR17 (rk− mk+) supE44 thi-1 recA1 gyrA96 relA1 Δ(argF-lacZYA)U169 φ80d lacZΔM15 | Invitrogen |
| Escherichia coli XL1-Blue MRA | Δ(mcrA) 183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 gyrA96 relA1 lac | Stratagene |
| Escherichia coli BL21(DE3) | F−dcm ompT hsdS (rB− mB−) gal λ (DE3) | Novagen |
| Escherichia coli BL21 AI | F−dcm ompT hsdS (rB− mB−) gal araB::T7RNAP-tetA | Invitrogen |
| Plasmids | ||
| pDrive | Kanr, cloning vector | QIAGEN |
| pGEM-T Easy | Apr, T-cloning vector | Promega |
| pCR-Blunt II-TOPO | Kanr, cloning vector | Invitrogen |
| pET15b | Ampr, expression vector | Invitrogen |
| pVLT31 | Tetr, expression vector | 10 |
| pSD1G3 | Supercos I carrying ca. 40-kb fragment of CHY-1 DNA including phnC | This study |
| pSD2C4 | Supercos I carrying ca. 40-kb fragment of CHY-1 DNA including phnA1aA2a-phnA2a | This study |
| pSD6 | pDrive containing phnA1a-phnA2a | This study |
| pSD8 | pET15b containing phnA1a-phnA2a | This study |
| pSD9 | pVLT31 containing phnA1a-phnA2a | This study |
| pDA11 | pDrive containing phnA1b | This study |
| pDB11 | pDrive containing phnA2b | This study |
| pDAB11 | pDrive containing phnA1b-phnA2b | This study |
| pED11 | pET9a carrying phnA1b-phnA2b | This study |
| pVED11 | pVLT31 containing phnA1b-phnA2b | This study |
| pSD11 | pDrive containing phnA3 | This study |
| pEBA3 | pET15b carrying phnA3 | This study |
| pDRA4 | pDrive containing phnA4 | This study |
| pEBA4 | pET15b carrying phnA4 | This study |
| pEB431 | pET15b carrying phnA4 and phnA3 | This study |
| pEB432 | pET15b carrying phnA4 and phnA3 in opposite orientation | This study |
| pSD13 | pCR-Blunt II-TOPO carrying a 3.06-kb fragment including phnA1a-phnA2a | This study |
| pSD14 | pSD13 carrying a phnA1a::Gmr deletion-insertion | This study |