TABLE 2.
Plasmids used in this study
| Plasmid | Description | Source or reference |
|---|---|---|
| pIJ2925 | pUC18 derivative | 2, 16 |
| pGM160Δ | pGM160 derivative, obtained by deletion of HindIII of pGM160 fragment carrying aac gene | 25 (gift of J. L. Pernodet) |
| pHP45 Ωaac | E. coli vector carrying Ωaac cassette | 5 |
| pSG100 | pIJ2925 derivative carrying PCR fragment encompassing ppk gene and 1-kb region located upstream of ppk | This study |
| pSG101 | pSG100 with EcoRI site of polylinker filled in with Klenow fragment | This study |
| pSG102 | pSG101 containing newly created EcoRI site | This study |
| pSG103 | Cloning of Ωaac resistance cassette from pHP45 Ωaac into newly created EcoRI site of pSG102 | This study |
| pSG104 | Cloning of insert of pSG103 into pGM160Δ | This study |
| pSG110 | Cloning of 3.2-kb PCR fragment carrying phoR and phoP genes of S. lividans into SmaI site of pIJ2925 | This study |
| pSG111 | Cloning of Ωaac resistance cassette from pHP45 Ωaac into NotI site of pSG110, filled in by Klenow fragment | This study |
| pSG112 | Cloning of insert of pSG111 into pGM160Δ | This study |