TABLE 1.
Key strains and plasmids in this study
| Strain or plasmid (reference) | Relevant characteristicsa |
|---|---|
| S. fradiae strains | |
| NRRL 18160 (11) | A54145 factor high producer |
| XH25 | NRRL 18160 Smr (rpsL K88R) |
| JR397 | XH25 ΔlptI |
| DA613 | XH25 ΔlptI::tsr |
| DA728 | XH25 ΔlptD |
| DA740 | XH25 ΔlptBCD |
| DA895 | XH25 ΔlptD ΔlptI::tsr |
| DA901 | XH25 ΔlptBCD ΔlptI::tsr |
| DA1187 | XH25 ΔlptEF-I::tsr |
| DA1077 | XH25 ΔlptI::tsr + ermEp* lptI (pDA1660) |
| DA1087 | XH25 ΔlptI::tsr + ermEp* dptI (pDA1658) |
| DA1096 | XH25 ΔlptI::tsr + lptI (pCV16) |
| DA1102 | XH25 ΔlptD + lptD-I (pDA2016) |
| DA1106 | XH25 ΔlptD + lptB-I (pDA2012) |
| DA1109 | XH25 ΔlptBCD + lptB-I (pDA2012) |
| DA1204 | XH25 ΔlptBCD ΔlptI::tsr + lptB-I (pDA2012) |
| DA1206 | XH25 ΔlptBCD ΔlptI::tsr + lptB-H (pDA2048) |
| DA1444 | XH25 ΔlptBCD ΔlptI::tsr + lptBC (pDA2040) + lptD-I (pDA2106) |
| DA1324 | XH25 ΔlptEF-I::tsr + lptEF-I (pJR153) |
| DA1674 | XH25 ΔlptEF-I::tsr + lptEF-P (pDA2054) |
| DA1676 | XH25 ΔlptEF-I::tsr + lptEF-I (pDA2002) |
| S. roseosporus strains | |
| UA431 (39) | Daptomycin nonproducer, ΔdptEF-J::tsr (“ΔdptEF-J::tsr”) |
| DA1155 | UA431 + lptEF-I (pDA2002) |
| DA1156 | UA431 + lptEF-I (pDA2002) |
| DA1160 | UA431 + pV89 (vector control) |
| S. ambofaciens strains | |
| BES2074 (44) | Spiramycin nonproducer, srmR7::Tn10 Nm |
| DA1163 | BES2074 + lptEF-I (pDA2002) |
| DA1164 | BES2074 + lptEF-I (pDA2002) |
| DA1168 | BES2074 + pV89 (vector control) |
| Plasmids | |
| pRHB538 (28) | rep(Ts) Am oriT |
| pJC06 | pRHB538::ΔlptD |
| pJC07 | pRHB538::ΔlptBCD |
| pJC11 | pRHB538::ΔlptI |
| pJC12 | pRHB538::ΔlptI::tsr |
| pDA1715 | pRHB538::ΔlptEF-I |
| pDA1716 | pRHB538::ΔlptEF-I::tsr |
| pDA1600 | pNEB193::Tsrr cassette |
| pDA1650 | Modified pIJ8660 (multiple cloning site) |
| pDA1652 | pDA1650::ermE*p |
| pDA1658 | pDA1652::ermE*p::lptI |
| pDA1660 | pDA1652::ermE*p::dptI |
| pDA1800 | pBR322::pECBAC1-HindIII + φC31 cassette + orf21 |
| pDA1803 | pBR322::pECBAC1-HindIII + φBT1 cassette + orf21 |
| pDA1811 | pBR322::pECBAC1 + fC31 cassette + ermE*p::lptB |
| pDA1814 | pBR322::pECBAC1 + φC31 cassette + ermE*p::lptD |
| pDA1834 | pBR322::pECBAC1 + φBT1 cassette + ermE*p::lptD |
| pCB01 | pECBAC1::lpt gene cluster |
| pJR153 | pCB01::φBT1 cassette at orf21 |
| pDA2002 | pCB01::φC31 cassette at orf21 |
| pDA2012 | pCB01::φC31 cassette + ermE*p::lptB |
| pDA2016 | pCB01::φC31 cassette + ermE*p::lptD |
| pDA1836 | pOJ260 + to terminator cassette |
| pDA2040 | pCB01::φC31 cassette + ermE*p::lptB, to terminator cassette in lptD (pDA2012) |
| pDA2048 | pCB01::φC31 cassette + ermE*p::lptB, to terminator cassette in lptJ (pDA2012) |
| pDA2054 | pCB01::φC31 cassette at orf21, to terminator cassette in lptI (pDA2002) |
| pDA2106 | pCB01::φBT1 cassette + ermE*p::lptD |
| pV89 | pNEB193::φC31 cassette |
| pCV16 (35) | pStreptoCos with partial lpt gene cluster including lptI, φC31 cassette |
a
lptD-I, lptDGHJKLMNPI; lptB-I, lptBCDGHJKLMNPI; lptB-H, lptBCDGH; lptEF-I, lptEFABCDGHJKLMNPI; lptEF-P, lptEFABCDGHJKLMNP; dptEF-J, dptEFABCDGHJ.