TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Characteristic(s) | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | Host for general cloning | Invitrogen |
| CC118 (λpir) | Host for general cloning | 9 |
| XL1-Blue MRF′ | Host for constructing the genomic library | Stratagene |
| S17-1 | Donor strain for conjugation between E. coli and Streptomyces | 26 |
| S17-1 (λpir) | Donor strain for conjugation between E. coli and Pseudomonas | 9 |
| BL21(DE3) | Donor strain for protein overexpression | Novagen |
| S. lavendulae strains | ||
| NRRL 11002 | Wild-type strain, SFM-A producing | NRRL |
| TL2001 | NRPS allele mutant of P1, SFM-A producing | This study |
| TL2002 | NRPS allele mutant of P2, SFM-A nonproducing | This study |
| TL2003 | ΔsfmB gene replacement mutant, SFM-A nonproducing | This study |
| TL2004 | TL2003 derivative containing pTL2007, expression of sfmB under the control of PermE* promoter, SFM-A producing | This study |
| TL2005 | orf(−1) gene disruption mutant, SFM-A producing | This study |
| TL2006 | orf(+1) gene disruption mutant, SFM-A producing | This study |
| P. fluorescens strains | ||
| FERM BP-14 | Wild-type strain, SAC-B producing | FERMa |
| TL2101 | FERM BP-14 containing pTL2011, expression of sfmO to sfmK under the control of the Ptac promoter | This study |
| TL2102 | FERM BP-14 containing pTL2014, expression of sfmO4 under the control of the Ptac promoter, aminated SFM-S producing | This study |
| Plasmids | ||
| pGEM-T Easy | E. coli subcloning vector | Promega |
| pGEM-7zf | E. coli subcloning vector | Promega |
| pSP72 | E. coli subcloning vector | Promega |
| pANT841 | E. coli subcloning vector | GenBank (accession no. AF438749) |
| pVLT33 | Heterologous expression vector in P. fluorescens A2-2 | 9 |
| pTL2001 | 1.2-kb PCR product of the NRPS gene (P1) in pGEM-T Easy | This study |
| pTL2002 | 1.2-kb PCR product of the NRPS gene (P2) in pGEM-T Easy | This study |
| pTL2003 | 0.8-kb PCR product of the NRPS RE gene (P3) in pGEM-T Easy | This study |
| pTL2004 | 1.2-kb EcoRI/SpeI internal fragment of pTL2001 in pOJ260 | This study |
| pTL2005 | 1.2-kb EcoRI/HindIII internal fragment of pTL2002 in pKC1139 | This study |
| pTL2006 | sfmB replacement construct in which sfmB was inactivated by insertion of ermE | This study |
| pTL2007 | 5.2-kb fragment containing sfmB under the control of PermE* in pTGV-4 | This study |
| pTL2008 | 1.4-kb PCR fragment containing sfmO3 to -K in pANT841 | This study |
| pTL2009 | 1.4-kb NdeI/HindIII fragment containing sfmO3 to -K in pET28a | This study |
| pTL2010 | 1.4-kb XbaI/HindIII fragment containing sfmO3 to -K in pVLT33 | This study |
| pTL2011 | 1.4-kb PCR fragment containing sfmO4 in pANT841 | This study |
| pTL2012 | 1.4-kb NdeI/HindIII fragment containing sfmO4 in pET28a | This study |
| pTL2013 | 1.4-kb XbaI/HindIII fragment containing sfmO4 in pVLT33 | This study |
| pTL2014 | 2.0-kb EcoRI/HindIII fragment that encodes the N terminus of SfmA in pSP72 | This study |
| pTL2015 | 5.5-kb fragment that encodes SfmA in pET28a | This study |
| pTL2016 | 3.4-kb fragment that encodes the truncated SfmA (C1-A1-PCP1) in pET28a | This study |
| pTL2017 | 0.6-kb EcoRI/PstI fragment that encodes the N terminus of SfmB in pSP72 | This study |
| pTL2018 | 0.5-kb EcoRI/HindIII fragment that encodes the C terminus of SfmB in pSP72 | This study |
| pTL2019 | 3.3-kb fragment that encodes SfmB in pET28a | This study |
| pTL2020 | 1.7-kb EcoRI/SphI fragment that encodes the N terminus of SfmC in pSP72 | This study |
| pTL2021 | 0.2-kb XbaI/HindIII fragment that encodes the C terminus of SfmC in pGEM-7zf | This study |
| pTL2022 | 4.5-kb fragment that encodes SfmC in pET28a | This study |
| pTL2023 | 0.3-kb PCR product encoding the internal fragment of orf(−1) in pSP72 | This study |
| pTL2024 | 0.4-kb PCR product encoding the internal fragment orf(+1) in pSP72 | This study |
| pTL2025 | 0.3-kb EcoRI/XbaI internal fragment of pTL2023 in pKC1139 | This study |
| pTL2026 | 0.4-kb EcoRI/XbaI internal fragment of pTL2024 in pKC1139 | This study |
| pTL2101 | S. lavendulae NRRL 11002 genomic library cosmid | This study |
| pTL2102 | S. lavendulae NRRL 11002 genomic library cosmid | This study |
| pTL2103 | S. lavendulae NRRL 11002 genomic library cosmid | This study |
a
FERM, Fermentation Research Institute, Agency of Industrial Science and Technology, Japan.