TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidDescriptionSource or reference
M. gryphiswaldense strains
    MSR-1 R3/S1Rifr Smr spontaneous mutant31
    ΔC::KanΔmamC::KnrThis study
    ΔCΔmamCThis study
    ΔC_CΔC(pAS35)This study
    ΔGFDCΔmamGFDCThis study
    ΔGFDC_MCS2ΔGFDC(pBBR1MCS-2)This study
    ΔGFDC_GFDCΔGFDC(pAS31)This study
    ΔGFDC_GΔGFDC(pAS32)This study
    ΔGFDC_FΔGFDC(pAS33)This study
    ΔGFDC_DΔGFDC(pAS34)This study
    ΔGFDC_CΔGFDC(pAS35)This study
    ΔGFDC_GDΔGFDC(pAS36)This study
    ΔGFDC_GCΔGFDC(pAS37)This study
    ΔGFDC_FDΔGFDC(pAS38)This study
    ΔGFDC_FCΔGFDC(pAS39)This study
    ΔGFDC_DCΔGFDC(pAS40)This study
    ΔGFDC_GFDΔGFDC(pAS41)This study
    ΔGFDC_GFCΔGFDC(pAS42)This study
    ΔGFDC_GDCΔGFDC(pAS43)This study
    ΔGFDC_FDCΔGFDC(pAS44)This study
Plasmids
    pBBR1MCS-2Knr lacZa17
    pK19mobsacBKnr sacB (modified from Bacillus subtilis) lacZ24
    pCM184Apr Knr18
    pCM157Tcr18
    pDC2pK19mobsacB with upstream and downstream mamGFDC cluster-flanking sequencesThis study
    pAG3pCM184 with upstream mamC-flanking sequence between MunI and NdeIThis study
    pAG4pAG3 with downstream mamC-flanking sequence between ApaI and SacIThis study
    pAS100pSP72 with 2.941-kb construct consisting of the 2,077-bp mamGFDC operon and 705-bp upstream and 159-bp downstream sequences between XhoI and SacI restriction sitesThis study
    pAS101pAS100 cut with NaeI and Eco47III and self-ligatedThis study
    pAS102pAS100 cut with PvuII and PsiI and self-ligatedThis study
    pAS103pAS100 cut with NruI and BfrBI, blunted, and self-ligatedThis study
    pAS104pAS100 cut with EcoRI and self-ligatedThis study
    pAS105pAS100 cut with NaeI and PsiI and self-ligatedThis study
    pAS106pAS100 cut with NaeI and BamHI, blunted, and self-ligatedThis study
    pAS107pAS100 cut with PvuII and BamHI, blunted, and self-ligatedThis study
    pAS109pAS105 cut with EcoRI and self-ligatedThis study
    pAS110pAS100 cut with PvuII and EcoRI, blunted, and self-ligatedThis study
    pAS111pAS101 cut with NruI and EcoRI, blunted, and self-ligatedThis study
    pAS112pAS104 cut with PvuII and PsiI and self-ligatedThis study
    pAS113pAS104 cut with NaeI and Eco47III and self-ligatedThis study
    pAS114pAS101 cut with NruI and BamHI, blunted, and self-ligatedThis study
    pAS31pBBR1MCS-2 with 2,941-bp XhoI-SacI fragment of pAS100, for mamGFDC expressionThis study
    pAS32pBBR1MCS-2 with 1,014-bp XhoI-SacI fragment from pAS110, for mamG expressionThis study
    pAS33pBBR1MCS-2 with 1,229-bp XhoI-SacI fragment from pAS111, for mamF expressionThis study
    pAS34pBBR1MCS-2 with 1,826-bp XhoI-SacI fragment from pAS109, for mamD expressionThis study
    pAS35pBBR1MCS-2 with 1,538-bp XhoI-SacI fragment from pAS106, for mamC expressionThis study
    pAS36pBBR1MCS-2 with 2,104-bp XhoI-SacI fragment from pAS112, for mamGD expressionThis study
    pAS37pBBR1MCS-2 with 1,819-bp XhoI-SacI fragment from pAS107, for mamGC expressionThis study
    pAS38pBBR1MCS-2 with 2,165-bp XhoI-SacI fragment from pAS113, for mamFD expressionThis study
    pAS39pBBR1MCS-2 with 2,038-bp XhoI-SacI fragment from pAS114, for mamFC expressionThis study
    pAS40pBBR1MCS-2 with 2,375-bp XhoI-SacI fragment from pAS105, for mamDC expressionThis study
    pAS41pBBR1MCS-2 with 2,401-kb XhoI-SacI fragment from pAS104, for mamGFD expressionThis study
    pAS42pBBR1MCS-2 with 2,265-bp XhoI-SacI fragment from pAS103, for mamGFC expressionThis study
    pAS43pBBR1MCS-2 with 2,668-kb XhoI-SacI fragment from pAS102, for mamGDC expressionThis study
    pAS44pBBR1MCS-2 with 2,722-kb XhoI-SacI fragment from pAS101, for mamFDC expressionThis study