TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristicsa | Reference or source |
|---|---|---|
| Alcanivorax borkumensis strains | ||
| SK2 | Type strain, wild type; TAG and WE producer | 68; DSM 11573 |
| SK2 algLΩSm | algL disruption mutant; Smr; derivative of SK2 | This study |
| SK2 algEΩSm | algE disruption mutant; Smr; derivative of SK2 | This study |
| SK2 MATEΩSm | MATE disruption mutant; Smr; derivative of SK2 | This study |
| Alcanivorax jadensis T9 | Type strain, wild type; TAG and WE producer | 13, 21; DSM 12178 |
| Acinetobacter baylyi ADP1 | TAG and WE producer | 31 |
| Escherichia coli strains | ||
| SM10 (λpyr) | Apr, Cmr, Kmr, pUTminiTn5Cm | 16 |
| TOP10 | F−mcrA Δ(mrr-hsdRMS mcrBC) rpcL nupG endA1 deoR φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK | Invitrogen |
| LE 392 | λ-rpsL(Smr) nupG F−hsdR574 (rK− mK+) supE44 supF58 lacY1 or Δ(lacIZY)6 galK2 galT22 metB1 trpR55 | Promega |
| XL1-Blue | recA1 endA gyrA96 thi1 hsdR17 (rK− mK+) supE44 relA1 λ−lac [F′ proAB lacIqZΔM15 Tn10(Tcr)] | 14 |
| S17-1 | recA1 thi1 hsdR17 (rK− mK+) proA tra (RP4) | 59 |
| Plasmids | ||
| pBluescript SK (−) | Plasmid used for subcloning DNA, Apr, lacPOZ′ | Stratagene |
| pUTminiTn5Cm | Suicide vector for mutagenesis with miniTn5, Cmr | 16 |
| pHC79 | Cosmid, Apr Tcr | 29 |
| pET19b | Apr, T7 promoter-based expression vector | Novagen |
| pET19b::algE | Derivative of pET19b containing the algE gene as 1,380bp HindIII fragment, Apr | This study |
| pET19b::algEΩSm | ΩSmr cassette cloned into SalI site of pET19b::algE, Apr Smr | This study |
| pUC19 | Cloning vector, Apr | 53 |
| pUC19::algL | Derivative of pUC19 containing the algL gene as 1,041-bp BamHI/HindIII fragment, Apr | This study |
| pUC19::algLΩSm | ΩSmr cassette cloned into NcoI site of pUC19::algL, Apr Smr | This study |
| pUC19::MATE | Derivative of pUC19 containing the MATE gene as 1,287-bp BamHI/HindIII fragment Apr | This study |
| pUC19::MATE:: ΩSm | ΩSmr cassette cloned into NcoI site of pUC19::algL, Apr Smr | This study |
| pSUP202 | Apr Cmr Tcr; ColE1 origin; mob site; unable to replicate in A. borkumensis | 59 |
| pSUP202:: algEΩSm | Fusion of pSUP202 and algEΩSm fragment of pET19b::algEΩSm via HindIII site; mob site, Cmr Apr Smr | This study |
| pSUP202:: algLΩSm | Fusion of pSUP202 and algLΩSm fragment of pUC19::algLΩSm via BamHI/HindIII sites; mob site, Cmr Apr Smr | This study |
| pSUP202:: MATEΩSm | Fusion of pSUP202 and MATEΩSm fragment of pUC19::MATEΩSm via BamHI/HindIII sites; mob site. Cmr Apr Smr | This study |
a
For abbreviations for genotypes of E. coli, see reference 7. Apr, ampicillin resistant; Cmr, chloramphenicol resistant; Smr, streptomycin resistant; Tcr, tetracycline resistant.