TABLE 1

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant phenotype or genotypeaReference or source
Strains
    M. smegmatis
        mc2155High-transformation-efficiency mutant of M. smegmatis ATCC 60753
        ΔaldR strainMSMEG_2660 (aldR) deletion mutant derived from M. smegmatis mc215561
        Δald strainMSMEG_2659 (ald) deletion mutant derived from M. smegmatis mc2155This study
        Δbd strainMSMEG_3233 (cydA) deletion mutant derived from M. smegmatis mc2155This study
        Δaa3 strainMSMEG_4268 (ctaC) deletion mutant derived from M. smegmatis mc2155This study
        Δbd Δald strainMSMEG_3233 (cydA) deletion mutant derived from M. smegmatis ΔaldThis study
    E. coli DH5αϕ80dlacZΔM15 ΔlacU169 recA1 endA1 hsdR17 supE44 thi1 gyrA96 relA162
Plasmids
    pKOTsHygr; pKO-based vector containing a temp-sensitive replication origin (pAL500Ts) and pUC ori61
    pNCHygr; promoterless lacZ63
    pMV306Kmr; integrative vector containing the int and attP sites of mycobacteriophage L5 for integration into the mycobacterial genome64, 65
    pNBV1Hygr; 5.8-kb vector derived from p16R166
    pKOTsΔaldpKOTs::0.81-kb BamHI-HindIII fragment containing ΔMSMEG_2659ald)This study
    pKOTsΔbdpKOTs::0.74-kb NotI-HindIII fragment containing ΔMSMEG_3233bd)This study
    pKOTsΔaa3pKOTs::0.94-kb BamHI-HindIII fragment containing ΔMSMEG_4268aa3)This study
    pALDLACZpNC::0.52-kb XbaI-ClaI fragment containing the ald promoter region of M. smegmatis mc215512
    pMV306ctaCpMV306 with 1.37-kb XbaI-HindIII fragment containing ctaC of M. smegmatis mc2155This study
    pMV306aldpMV306 with 1.55-kb XbaI-HindIII fragment containing ald of M. smegmatis mc2155This study
    pNBV1cydApNBV1 with 1.92-kb XbaI-HindIII fragment containing cydA of M. smegmatis mc2155This study
  • a Antibiotic resistance is indicated by abbreviation (Hyg, hygromycin; Km, kanamycin).