Table 1. Strains, cosmids and plasmids used in this study.
| Strain | Reference | |
|---|---|---|
| S4 | Wild type Streptomyces albus S4 | Barke et al., 2010 |
| S4 ΔantA | S4 antA null mutant; AprR | This study |
| S4 antA attBΦBT1::pIJ10257-antA | Complemented antA null mutant; antA transcription driven by the ermE* promoter; AprR, HygR | This study |
| S4 ΔantA attBΦBT1::pAU3-45-antA-AA | Complemented antA null mutant; antA driven its native promoter; AprR TspR | This study |
| S4 ΔantA attBΦBT1::pAU3-45-antA-DD | Complemented antA null mutant with mutations A172D A173D with antA driven its native promoter; AprR TspR | This study |
| S4 ΔantC | S4 antC null mutant; AprR | This study |
| S4 ΔSTRS4_02194 | S4 STRS4_02194 null mutant; AprR | This study |
| S4 ΔSTRS4_02195 | S4 STRS4_02195 null mutant; AprR | This study |
| S4 ΔSTRS4_02212 | S4 STRS4_02212 null mutant; AprR | This study |
| S4 ΔSTRS4_02213-02217 | S4 STRS4_02213-02217 null mutant; AprR | This study |
| Escherichia coli | ||
| BL21 | Host for heterologous protein expression | Novagene |
| BW25113 | Host for REDIRECT PCR targeting system | Gust et al., 2003 |
| DHM1 | Host for Cya bacterial two hybrid system | Karimova et al., 1998 |
| ET12567 | Non-methylating host for transfer of DNA into Streptomyces spp. (dam, dcm, hsdM); CamR | MacNeil et al., 1992 |
| TOP10 | General cloning host | Invitrogen |
| VCS257 | Host strain for Gigapack III XL phage | Agilent Technologies |
| Cosmids | ||
| Supercos1 | Cosmid backbone for S. albus S4 cosmids; AmpR, KanR | Stratagene |
| Cosmid 213 | Supercos1 derviative containing the entire antimycin gene cluster; AmpR, KanR | This study |
| Cosmid 213 ΔantB-flp | Cosmid 213 derivative containing an 81 bp scar in place of the antB gene | This study |
| Cosmid 456 | Supercos1 derviative containing a portion of the antimycin gene cluster; AmpR, KanR | This study |
| Plasmids | ||
| pCRII-TOPO | Cloning vector for PCR products; AmpR, KanR | Invitrogen |
| pAU3-45 | pSET152 derivative, integrates into ΦC31 attB site in Streptomyces; AprR TspR | Bignell et al., 2005 |
| pAU3-45-antA-AA | pAU3-45 derivative containing the antA gene, an 81 bp ‘scar’ in place of antB, and 270 bp upstream of antB cloned into the EcoRI site | This study |
| pAU3-45-antA-DD | pAU3-45 derivative containing the antA gene encoding A172D and A173D mutations, an 81 bp ‘scar’ in place of antB, and 270 bp upstream of antB, cloned into the EcoRI site | This study |
| pET28a | Protein expression vector; KanR | Novagene |
| pET28a-antA | pET28a derivative containing the antA coding sequence cloned into the NdeI and HindIII sites | This study |
| pGEM-T Easy | Cloning vector for PCR products; AmpR | Promega |
| pIJ773 | PCR template for aac3(IV)+ oriT cassette used in REDIRECT PCR targeting system | Gust et al., 2003 |
| pIJ790 | Encodes lambda RED recombination machinery induced by arabinose; CamR | Gust et al., 2003 |
| pIJ10700 | PCR template for hygR cassette used in REDIRECT PCR targeting system | Gust et al., 2003b |
| pIJ10257 | pMS81 derivative containing ermE*p, integrates into the ΦBT1 attB site in Streptomyces; HygR | Hong et al., 2005 |
| pIJ10257-antA | pIJ10257 derivative containing the antA coding sequence cloned into the NdeI-HindIII sites | This study |
| pUZ8002 | Encodes conjugation machinery for mobilization of plasmids from E. coli to Streptomyces; KanR | MacNeil et al., 1992 |
Notes.
- Amp
- ampicillin
- Apr
- apramycin
- Hyg
- hygromycin
- Kan
- kanamycin
- Cam
- chloramphenicol