TABLE 1.
Strains and plasmids used or constructed in this work
| Strain or plasmid | Relevant genotype or characteristics | Reference and/or source |
|---|---|---|
| E. coli strains | ||
| CC118(RK600) | Helper strain for triparental mating | 6 |
| DH5α | Cloning strain | Invitrogen |
| JS200(pEP Pol I) | Mutator strain | Addgene; 3 |
| XL1-Red | Mutator strain | Invitrogen |
| P. putida strains | ||
| GPo12(pGEc47ΔB) | GPo1 cured of the OCT plasmid, transformed with pGEc47ΔB; contains all genes necessary for growth on alkanes, except for the alkane hydroxylase gene | 26 |
| Palk1 | GPo12(pGEc47ΔB) adapted to growth on gaseous alkanes with pCom10_alkB | This study |
| Pcyp1 | GPo12(pGEc47ΔB) adapted to growth on gaseous alkanes with pCom8_cyp153A6 | This study |
| Pcyp2 | Pcyp1 further adapted to growth on gaseous alkanes with pCom8*_cyp153A6-BMO1 | This study |
| Plasmids | ||
| pCom8 | Broad-host-range expression vector with PalkB; GmroriT alkS | 25 |
| pCom8* | pCom8 vector adapted for improved complementation for small-chain-length alkane-dependent host growth | This study |
| pCom10 | Broad-host-range expression vector with PalkB; KanroriT alkS | 25 |