Table 3.
Strains and plasmids used
| Strain/plasmid | Description/ function | Source |
|---|---|---|
| Strains | ||
| S. ghanaensis ATCC14672 | Wild type (WT) moenomycin producer | ATCC |
| S. ghanaensis GT1 | WT, moeGT1 inactivation; Amr Kmr | Ostash et al. (2007) |
| S. ghanaensis dB4 | WT, ΔmoeB4 deletion mutant; Amr | This study |
| S. ghanaensis dH5 | WT, AmoeH5 deletion mutant; Amr | This study |
| S. lividans TK24 | Host for moenomycin production and rMoeH5 protein expression; Smr | Kieser et al. (2000) |
| S. lividans TK24 moeno38-5+ | TK24 carrying cosmid moeno38-5 | Ostash et al. (2009) |
| S. lividans TK24 moeno38-6+ | TK24 carrying cosmid moeno38-6 | Ostash et al. (2009) |
| S. albus J1074 | Host for moenomycin production | Makitrinskyy et al. (2010) |
| S. albus J1074 moeno38-6+ | J1074 carrying cosmid moeno38-6 | This study |
| E. coli XL1 Blue MR | Host strain for DNA cloning | Stratagene |
| E. coli BL21(DE3) pLysS | Host for MoeF5 protein expression | Novagen |
| E. coli ET12567 | Host for DNA conjugation | Kieser et al. (2000) |
| E. coli BW25113 | Host for DNA recombineering | Gust (2009) |
| Cosmids | ||
| moeno38 | SuperCos1-based, carries entire moe cluster 1 except moeR5moeS5, KmrApr | Ostash et al. (2007) |
| moeno38-1 | moeno38, replacement of neo with intφC31-oriTRK2-hyg cassette, Hyr | Ostash et al. (2007) |
| moeno38-5 | moeno38-1 ΔmoeA4ΔmoeB4, HyrKmr | Ostash et al. (2009) |
| moeno38-6 | moeno38-1 ΔmoeH5 | Ostash et al. (2009) |
| moeno38-dH5 | moeno38 ΔmoeH5 | This study |
| moeno5 | SuperCos1-based, carries entire moe cluster 2 | Ostash et al. (2007) |
| moeno5-dB4 | moeno5, ΔmoeB4 | This study |
| Plasmids | ||
| pSOK804 | VWB-based integrative vector, Amr | Sekurova et al. (2004) |
| pOOB64b | pSOK804 carrying entire moe cluster 2 | Ostash et al. (2009) |
| pOOB64bd | pOOB64b, ΔmoeB4 deletion, AmrHyr | This study |
| pKC1139E | pKC1139 carrying ermEp, Amr | Ostash et al. (2007) |
| pOOB47a | pKC1139E, ermEp-moeH5 fusion, Amr | This study |
| pOOB47h | pKC1139E, ermEp-moeH5 fusion, Hyr | This study |
| pOOB48a | pKC1139E, ermEp-moeF5 fusion, Amr | Ostash et al. (2009) |
| pOOB83e | pKC1139E, ermEp-moeH5-6His, Amr | This study |
| pCXL5 | pET24a, moeF5 for production of C-terminally 6His-tagged MoeF5, Kmr | D. Perlstein |
| pHYGl | pUC19 carrying hyg gene, AprHyr | J. Salas |
| pIJ773 | Source of aac(3)IV-oriT cassette | Gust (2009) |
| pUZ8002 | Expression of proteins for plasmid conjugation in trans, Kmr | Kieser et al. (2000) |
| pIJ790 | Expression of proteins Bet, Exo and Gam for recombineering, Cmr | Gust (2009) |
Amr, apramycin resistant; Kmr , kanamycin resistant; Smr, streptomycin resistant; Apr, ampicillin resistant; Hyr , hygromycin resistant; Cmr, chloramphenicol resistant.