TABLE 1.
Strains, plasmids, and cosmids used in this study
| Strain, plasmid, or cosmid | Description | Reference or source |
|---|---|---|
| S. aureochromogenes | ||
| CXR14 | YB172 (industrial POL producer) derivative with the entire POL gene cluster deleted | 13 |
| CXR14::pCHW201 | Strain CXR14 containing pCHW201 | This study |
| CXR14::pCHW201/ΔpolG | Strain CXR14 containing pCHW201/ΔpolG | This study |
| CXR14::pJTU2463b | Strain CXR14 containing pJTU2463b | This study |
| E. coli | ||
| DH10B | F− mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80dlacZΔM15 ΔlacX74 deoR recA1 endA1 araΔ139 D(ara leu)7697 galU galK λ− rpsL nupG | Gibco-BRL |
| BW25113/pIJ790 | λ Red (gamma beta exo) cat araC rep101 | 18 |
| Rosetta(DE3)/pLysS | F− ompT hsdSB(rB− mB−) gal dcm λ(DE3 [lacI lacUV5-T7 gene1 ind1 sam7 nin5]) pLysS (Cmlr) | Novagen |
| ET12567(pUZ8002) | F− dam-13::Tn9 dcm-6 hsdM hsdR recF143 zjj-202::Tn10 galK2 galT22 ara14 pacY1 xyl-5 leuB6 thi-1 pUZ8002 | 31 |
| Plasmids | ||
| pJTU2463b | int aac(3)IV oriT RK2 phiC31 attP | This study |
| pET28a | neo reppMB1 T7 promoter | Novagen |
| pJTU4774 | The cosmid with XbaI and SpeI sites infilled, containing the entire POL gene cluster | This study |
| pCHW201 | pJTU4774 derivative with polF in frame deleted via PCR-targeting technology | This study |
| pCHW201/ΔpolG | pCHW201 derivative with polG in frame deleted via PCR-targeting technology | This study |
| pET28a/polG | pET28a derivative carrying an NdeI-EcoRI fragment containing the optimized polG | This study |
| pET28a/polGK110A | pET28a/polG derivative with the K110A mutation | This study |
| pET28a/polGK150A | pET28a/polG derivative with the K150A mutation | This study |
| pET28a/polGE188A | pET28a/polG derivative with the E188A mutation | This study |
| pET28a/polGE267A | pET28a/polG derivative with the E267A mutation | This study |
| pET28a/polGE280A | pET28a/polG derivative with the E280A mutation | This study |