Table 3.
Constructs and strains used in the study
| Description | |
|---|---|
| Constructs | |
| pENTRCmr | pENTR chloramphenicol cloned in KpnI and SnaBI amplified from pVR1 |
| pNit1 (84) | Nitrile-inducible vector kanr |
| pNitchl | pNit1 vector modified by inserting cmr gene |
| pST-KT (38) | Anhydrotetracyclin-inducible vector (ref) kanr |
| pJV53 (41) | Plasmid encoding phage phage Che9c 60 and 61 genes under acetamide-inducible promoter; kanr |
| pNit-ET (85) | Plasmid encoding phage Che9c 60 and 61 genes cloned in pNit1; kanr |
| Strains | |
| DH5α | E. coli strain used for cloning experiments |
| Msm | WT M. smegmatis mc2155 strain |
| Mtb | WT H37Rv M. tuberculosis strain |
| Mtb::Vc | Mtb electroporated with pST-KT; kanr |
| Mtb::rodA | Mtb electroporated with pST-KT-RodAMtb; kanr |
| Mtb::pbpA | Mtb electroporated with pST-KT-PbpAMtb; kanr |
| Mtb::pNitET | Mtb electroporated with pNit-ET; kanr |
| MtbΔr | rodAMtb gene replacement mutant; hygr |
| MtbΔp | pbpAMtb gene replacement mutant; hygr |
| MtbΔrp | rodAMtb-pbpAMtb gene replacement mutant; hygr |
| MtbΔr::r | MtbΔr complemented with pNitchl-RodAMtb; hygr, cmr |
| MtbΔp::p | MtbΔp complemented with pNitchl-PbpAMtb; hygr, cmr |
| MtbΔr::r | MtbΔr complemented with pST-KT-RodAMtb; hygr, kanr |
| MtbΔr::rT463A | MtbΔr complemented with pST-KT-RodAMtb-T463A; hygr, kanr |
| MtbΔr::rT463E | MtbΔr complemented with pST-KT-RodAMtb-T463E; hygr, kanr |
| Msm::pJV53 | Msm electroporated with pJV53 plasmid; kanr |
| MsmΔr | Msm rodA gene replacement mutant; hygr |
| MsmΔp | Msm pbpA gene replacement mutant; hygr |
| MsmΔrp | Msm rodA-pbpA gene replacement mutant; hygr |
| Msm::pN | Msm electroporated with pNitchl; cmr |
| Msm::pN-r | Msm electroporated with pNitchl-RodAMtb; cmr |
| Msm::pN-p | Msm electroporated with pNitchl-PbpA; cmr |
| MsmΔp::pMtb | MsmΔp complemented with pST-KT-PbpAMtb; hygr, kanr |
| MsmΔp::p | MsmΔp electroporated with pNit-PbpA; hygr, kanr |
| MsmΔp::pS281A | MsmΔp electroporated with pNit-PbpAS281A; hygr, kanr |
| MsmΔp::pK424G | MsmΔp electroporated with pNit-PbpAK424G; hygr, kanr |
| MsmΔr::rMtb | MsmΔr complemented with pST-KT-RodAMtb; hygr, kanr |
| MsmΔr::rMsm | MsmΔr complemented with pST-KT-RodAMtb; hygr, kanr |
| MsmΔr::ftsW | MsmΔr complemented with pST-KT-FtsWMtb; hygr, kanr |
| MsmΔr::mviN | MsmΔr complemented with pST-KT-MviNMtb; hygr, kanr |
| MsmΔp::pMtb | MsmΔp complemented with pST-KT-PbpAMtb; hygr, kanr |
| MsmΔr::rMtb-W175R | MsmΔr complemented with pST-KT-RodAMtb-W175R; hygr, kanr |
| MsmΔr::rMtb-D343R | MsmΔr complemented with pST-KT-RodAMtb-D343R; hygr, kanr |
| MsmΔr::rMtb-DM | MtbΔr complemented with pST-KT-RodAMtb-W175R -D343R; hygr, kanr |
| MsmΔr::rMsm-W176R | MtbΔr complemented with pST-KT-RodAMsm-W176R; hygr, kanr |
| MsmΔr::rMsm-D344R | MtbΔr complemented with pST-KT-RodAMsm-D344R; hygr, kanr |
| MsmΔr::rMsm-DM | MtbΔr complemented with pST-KT-RodAMsm-W176R -D344R; hygr, kanr |