Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mycobacterium smegmatis) | Mycobacterium smegmatis mc2155 | Wild type Mycobacterium smegmatis | ||
| Strain, strain background (Mycobacterium smegmatis) | KG26 | This work | mc2155 ∆MSMEG_0317 L5::pTetO-MSMEG_0317 | Parental swap strain |
| Strain, strain background (Mycobacterium smegmatis) | HR342 | Rego et al., 2017 | mc2155 ∆MSMEG_4265 | LamA knockout strain |
| Strain, strain background (Mycobacterium smegmatis) | KG97 | This work | mc2155 ∆MSMEG_0317 L5::pTetO-MSMEG_0317-myc / pTetR | Strain used for the anhydrotetracyline-inducible depletion of PgfA |
| Strain, strain background (Mycobacterium smegmatis) | KG81 | This work | mc2155 ∆MSMEG_0317 pL5::pNative-MSMEG_0317 -mRFP-myc | Strain used for determining PgfA localization |
| Strain, strain background (Mycobacterium smegmatis) | HR388 | Rego et al., 2017 | mc2155 L5::pG-MCK-ptb21-eGFP-wag31 tweety::ptb21-ftsZ-mCherry2B | Strain used to determine localization of FtsZ and Wag31 |
| Strain, strain background (Mycobacterium smegmatis) | CMG184 | This work | mc2155 tw::ptb21 mmpL3TB-msfGFP | Strain used to determine localization of MmpL3 |
| Strain, strain background (Mycobacterium smegmatis) | CMG530 | This work | mc2155 ∆MSMEG_0317 L5::pTetO-MSMEG_0317-3x- FLAG tw::ptb21-mmpL3TB-msfGFP | Strain used for co-immunoprecipitation of MmpL3 and PgfA |
| Strain, strain background (Mycobacterium smegmatis) | KG167 | This work | mc2155 ∆MSMEG_0317 L5::pTetO-MSMEG_0317-3x-FLAG | Control strain used for co-immunoprecipitation of MmpL3 and PgfA |
| Strain, strain background (Mycobacterium smegmatis) | CMG184 | This work | mc2155 tw::ptb21-mmpL3TB-msfGFP | Strain used to determine localization of MmpL3 |
| Strain, strain background (Mycobacterium smegmatis) | CMG481 | This work | mc2155 ∆lamA::zeo L5:: pNative-lamA-strep tw:: ptb21-mmpL3TB-msfGFP | Strain used to determine localization of MmpL3 in a ∆lamA background with LamA complemented under its native promoter |
| Strain, strain background (Mycobacterium smegmatis) | CMG557 | This work | mc2155 L5::pTetO-MSMEG_0317-myc | Strain used for measuring polar growth in a PgfA merodiploid background |
| Strain, strain background (M. smegmatis) | KG175 | This work | mc2155 ∆lamA L5::pTetO- MSMEG_0317-myc | Strain used for measuring polar growth in a PgfA merodiploid and LamA knockout background |
| Strain, strain background (Mycobacterium smegmatis) | CMG585 | This work | mc2155 L5::pTetO-mmpL3TB-myc | Strain used for measuring polar growth in an MmpL3 merodiploid background |
| Strain, strain background (Mycobacterium smegmatis) | CMG587 | This work | mc2155 ∆lamA L5:: pTetO-mmpL3TB-myc | Strain used for measuring polar growth in an MmpL3 merodiploid and LamA knockout background |
| Strain, strain background (Mycobacterium smegmatis) | KG154 | This work | mc2155 ∆lamA ∆MSMEG_0317 L5::pNative-MSMEG_0317 -mRFP-myc | Strain used for PgfA localization in LamA knockout background |
| Strain, strain background (Mycobacterium smegmatis) | KG157 | This work | mc2155 ∆lamA ∆MSMEG_0317 L5::pNative-MSMEG_0317 -mRFP-myc L5::pNative-lamA-strep | Strain used for PgfA localization in a background where lamA was complemented in LamA knockout |
| Strain, strain background (Mycobacterium smegmatis) | CMG206 | This work | mc2155 tw::ptb21-msfGFP-MurJTB | Strain used for localization of MurJ |
| Strain (Mycobacterium smegmatis) | CMG199 | This work | mc2155 ∆lamA tw::ptb21-msfGFP-murJTB | Strain used for localization of MurJ in a LamA knockout background |
| Strain, strain background (Mycobacterium smegmatis) | CMG485 | This work | mc2155 ∆lamA::zeo L5::pNative-LamA-strep tw::ptb21-msfGFP-MurJTB |
Strain used for localization of MurJ in a LamA knockout background where LamA was complemented under its native promoter |
| Strain, strain background (Mycobacterium smegmatis) | KG244 | This work | mc2155 L5::pLJR962-mmpL3-sgRNA | CRISPR-inducible MmpL3 depletion strain |
| Strain, strain background (Mycobacterium smegmatis) | KG289/ CMG549 |
This work | mc2155 L5::pLJR962- MSMEG_0317-sgRNA tw::pNative- MSMEG_0315-strep | CRISPR-inducible PgfA depletion strain |
| Strain, strain background (Mycobacterium smegmatis) | KG47 | This work | mc2155 ∆MSMEG_0317 L5::pTetO-MSMEG_0317-mGFPmut3-myc | C-terminal GFP tagged PgfA strain |
| Strain, strain background (Mycobacterium smegmatis) | KG73 | This work | mc2155 ∆MSMEG_0317 L5::pTetO-mGFP-mut3- MSMEG_0317-myc | N-terminal GFP tagged PgfA strain |
| Strain, strain background (Mycobacterium smegmatis) | KG167 | This work | mc2155 ∆MSMEG_0317 L5::pTetO- MSMEG_0317-3xFLAG | Strain carrying MSMEG_0317-3×-FLAG, used for TMM pulldown |
| Strain, strain background (Mycobacterium smegmatis) | KG60 | This work | mc2155 /pTetOR- MSMEG_0317-myc | Strain carrying tetracycline inducible overexpression plasmid for PgfA |
| Strain, strain background (Mycobacterium smegmatis) | KG286 | This work | mc2155 Δmsmeg_0317 L5::pTetO- MSMEG_0317-ΔC | Strain carrying msmeg_0317 without c-terminal cytoplasmic tail |
| Strain, strain background (Mycobacterium smegmatis) | HR404 | This work | mc2155 ΔMSMEG_6398 | Antigen 85A KO strain used in TLC control experiments |
| Recombinant DNA reagent | KG147 (plasmid) | This work | DH5α / pL5-pTetO-Ncgl2760-myc |
Plasmid carrying corynebacterial ncgl2760 gene |
| Recombinant DNA reagent | KG284 (plasmid) | This work | DH5α / pL5-pTetO-Ncgl2760-MSMEG_0317-C-term |
Plasmid carrying corynebacterial ncgl2760 gene fused to cytosolic C-terminal of msmeg_0317 gene |
| Recombinant DNA reagent | CMG541 (plasmid) | This work | Dh5a / pTweety-pNative MSMEG_0315-strep | Plasmid carrying mycobacterial gene MSMEG_0315 with a C-terminal strep tag |