TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| Chromobacterium violaceum | ||
| no. 968 (FERM BP-1968) | Wild-type strain, FK228 producing, Apr Thior | IPODb |
| Escherichia coli | ||
| DH5α | General cloning host | Lab stock |
| S17-1 | Host strain for interspecies conjugation | Lab stock |
| BL21(DE3) | Host strain for heterologous gene expression | Novagen |
| SW01 | BL21(DE3) harboring cosmid 18 | This work |
| SW02 | BL21(DE3) harboring cosmid 18 and pBMTL-3-depR | This work |
| SW03 | BL21(DE3) harboring cosmid 18, pBMTL-3-depR, and pCDFDuet-1-sfp | This work |
| SW04 | BL21(DE3) harboring cosmid 18, pBMTL-3-depR, and pCDFDuet-1-acpS | This work |
| SW05 | BL21(DE3) harboring cosmid 18, pBMTL-3-depR, and pCDFDuet-1-fabD1 | This work |
| SW06 | BL21(DE3) harboring cosmid 18, pBMTL-3-depR, and pCDFDuet-1-fabD2 | This work |
| SW07 | BL21(DE3) harboring cosmid 18, pBMTL-3-depR, and pCDFDuet-1-sfp-fabD1 | This work |
| SW08 | BL21(DE3) harboring cosmid 18, pBMTL-3-depR, and pCDFDuet-1-sfp-fabD2 | This work |
| SW09 | BL21(DE3) harboring cosmid 18, pBMTL-3-depR, and pCDFDuet-1-acpS-fabD1 | This work |
| SW10 | BL21(DE3) harboring cosmid 18, pBMTL-3-depR, and pCDFDuet-1-acpS-fabD2 | This work |
| Plasmids | ||
| pGEM-3Zf | Apr, general cloning vector | Promega |
| pGEM-T Easy | Apr, general cloning vector | Promega |
| Cosmid 18 | Apr Kanr, cosmid clone containing the FK228 biosynthetic gene cluster (dep) and flanking DNAs, shotgun sequenced | 9 |
| pBMTL-3 | Cmr, pBBR1 ori, broad-host-range vector | 19 |
| pBMTL-3-depR | Cmr, depR (with ribosome-binding site from pET29a) cloned into pBMTL-3 | 23 |
| pCDFDuet-1 | Smr, CDF ori, dual expression vector | Novagen |
| pCDFDuet-1-sfp | sfp cloned into the MCS1 region of pCDFDuet-1 | This study |
| pCDFDuet-1-acpS | acpS cloned into MCS1 region of pCDFDuet-1 | This study |
| pCDFDuet-1-fabD1 | fabD1 cloned into MCS2 region of pCDFDuet-1 | This study |
| pCDFDuet-1-fabD2 | fabD2 cloned into MCS2 region of pCDFDuet-1 | This study |
| pCDFDuet-1-sfp-fabD1 | sfp cloned into MCS1, fabD1 cloned into MCS2 of pCDFDuet-1 | This study |
| pCDFDuet-1-sfp-fabD2 | sfp cloned into MCS1, fabD2 cloned into MCS2 of pCDFDuet-1 | This study |
| pCDFDuet-1-acpS-fabD1 | acpS cloned into MCS1, fabD1 cloned into MCS2 of pCDFDuet-1 | This study |
| pCDFDuet-1-acpS-fabD2 | acpS cloned into MCS1, fabD2 cloned into MCS2 of pCDFDuet-1 | This study |
a
Apr, ampicillin resistance; Cmr, chloramphenicol resistance; Kanr, kanamycin resistance; Smr, streptomycin resistance; Thior, thiostrepton resistance; MCS, multiple cloning site.
b
IPOD, International Patent Organism Depositary, Tsukuba, Japan.