TABLE 2

Bacterial strains and plasmids used in this study

Strain or plasmidDescriptionReference or source
Strains
    S. coelicolor
        M145Wild-type, SCP1 SCP245
        ΔmacRmacR::aac(3)IVThis study
        C-ΔmacRΔmacR complemented with pCom-2120This study
        C-ΔmacR-FlagΔmacR complemented with pCom-2120-FLAGThis study
        ΔmacSmacS::aac(3)IVThis study
        C-ΔmacSΔmacS complemented with pCom-2121This study
        ΔmacRSmacRS::aac(3)IVThis study
        C-ΔmacRSΔmacRS complemented with pCom-2120/21SCOThis study
        C-ΔmacRSSAVΔmacRS complemented with pCom-6081/82SAVThis study
        C-ΔmacRSSVENΔmacRS complemented with pCom-1780/81SVENThis study
        ΔmmpAmmpA::aac(3)IVThis study
        C-ΔmmpAΔmmpA complemented with pCom-6728This study
        ΔmmpBmmpB::aac(3)IVThis study
        C-ΔmmpBΔmmpB complemented with pCom-4924This study
        ΔmmpCmmpC::aac(3)IVThis study
        C-ΔmmpCΔmmpC complemented with pCom-4011This study
    E. coli
        NovaBlueGeneral cloning strainNovagen
        Transetta BL21 (DE3)Strain used for protein expressionSangon
        ET12567(pUZ8002)Strain used for conjugation between E. coli and Streptomyces45
        BW25113Strain used for PCR targeting48
Plasmids
    pMD18-TGeneral cloning vectorTakara
    pCR-BLUNTGeneral cloning vectorInvitrogen
    pMS82Streptomyces integrative vector with hygromycin resistance49
    pEX-2120MacR expression plasmidThis study
    pCom-2120pMS82 with 526-bp upstream sequence and 648-bp coding sequence of macRThis study
    p2120-FLAGpCom-2120 with a 3×FLAG epitope inserted before the stop codon of macRThis study
    pCom-2121pMS82 with 284-bp upstream sequence and 1,242-bp coding sequence of macSThis study
    pCom-4011pMS82 with 359-bp upstream sequence and 474-bp coding sequence of mmpCThis study
    pCom-4924pMS82 with 386-bp upstream sequence and 129-bp coding sequence of mmpBThis study
    pCom-6728pMS82 with 260-bp upstream sequence and 147-bp coding sequence of mmpAThis study
    pCom-2120/21SCOpMS82 with 1,956-bp coding sequence of macRS and 245-bp upstream sequence of macSThis study
    pCom-6081/82SAVpMS82 with 1,907-bp coding sequence of sav6081-82 and 617-bp upstream sequence of sco6081This study
    pCom-1780/81SVENpMS82 with 1,892-bp coding sequence of sav6081-82 and 333-bp upstream sequence of sco6081This study