TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| S. paucimobilis | ||
| SYK-6 | Wild type; Nalr Smr | 9 |
| DLC | Mutant derivative of SYK-6; Kmr gene insertion mutant of ligC; Nalr Smr Kmr | This study |
| P. putida PpY1100 | Nalr Smr | 9 |
| E. coli | ||
| JM109 | recA1 supE44 endA1 hsdR17 gyrA96 relA1 thi Δ(lac-proAB) F′[traD36 proAB+lacIqlacZΔM15] | 42 |
| HB101 | supE44 hsdS20(rB− mB−) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 3 |
| BL21(DE3) | hsdS gal(λcIts857 ind1 Sam7 nin5 lacUV5-T7 gene 1) | 37 |
| Plasmids | ||
| pUC18 and pUC19 | Cloning vectors; Apr | 42 |
| pBluescript II SK(+) | Cloning vectors; Apr | 36 |
| pET21(+) | Expression vector; Apr T7 promoter | Novagen |
| pKT230 | Broad-host-range vector; Kmr | 2 |
| pVK100 | Broad-host-range cosmid; Kmr Tetr | 11 |
| pBBR122 | Broad-host-range vector; Kmr Cmr | MoBiTec |
| pK19mobsacB | oriT sacB Kmr | 34 |
| pUC4K | Source of Kmr cassette; Apr Kmr | 40 |
| pVA01 | pKT230 with a SYK-6 10.9-kb insert carrying ligABC and ligI | 9 |
| pVAD2 | Deletion plasmid of pVA01; a SalI fragment near the 3′ terminus of the insert was deleted | This study |
| pVAD4 | Deletion plasmid of pVA01; SalI fragments in the middle of the insert were deleted | 23 |
| pHN139F | pUC18 with a 10.5-kb EcoRI fragment of pVA01 carrying ligAB, part of ligC, and ligI | 23 |
| pHN139R | pUC18 carrying the same fragment as pHN139F in the opposite direction | 23 |
| pHNC2 | pET21(+) with a 1.0-kb SmaI-EcoRI fragment carrying part of ligC | This study |
| pSM21 | pET21(+) with a 1.4-kb SmaI-EcoRI fragment carrying ligC | This study |
| pAB16 | pBluescript II SK(+) with a 1.6-kb XbaI-XhoI fragment carrying ligAB | This study |
| pABC25 | pBluescript II SK(+) with a 2.5-kb XbaI-EcoRI fragment carrying ligAB and part of ligC | This study |
| pABC25K | pABC25 with insertion of the Kmr gene of pUC4K replacing a 0.2-kb BsmI fragment | This study |
| pLCD1 | pK19mobsacB with a 3.5-kb XbaI-EcoRI fragment of pABC25K | This study |
| pBBRC1220 | pBBR122 with a 1.4-kb SmaI-HindIII fragment carrying ligC of pSM21 | This study |
| pBBRC1221 | pBBRC1220 with an 0.4-kb PvuII fragment carrying the lac promoter of pUC19 in the SmaI site | This study |