TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Relevant genotype and/or information | Reference or source |
|---|---|---|
| Strains | ||
| E. coli DH10B | F−araD139 Δ(ara leu)7697 ΔlacX74 galU galK rpsL deoR φ80dlacZΔM15 endA1 nupG recA1 mcrA Δ(mrr hsdRMS mcrBC) | 11 |
| M. smegmatis | ||
| mc2155 | Wild type, ept-1 | 23 |
| DM5 | mc2155 harboring pDM2 (Δalr::aphA-3 Kanr Gmr) | This study |
| DM22 | mc2155 Δalr::aphA-3 Kanr | This study |
| DM30 | DM22 harboring pDM3; Hygr | This study |
| Plasmids | ||
| pPR23 | E. coli-mycobacterial shuttle vector; oriM(Ts) sacB Gmr | 19 |
| pUHA267 | E. coli-mycobacterial shuttle vector; Hygr | AgResearch, Wallaceville, New Zealand |
| pDM2 | pPR23 harboring Δalr::aphA-3; Kanr Gmr | This study |
| pDM3 | pUHA267 harboring the wild-type alr and upstream promoter region; Hygr | This study |
| Primers | ||
| AlrKO1 | 5′-AAATTTGGATCCGACAAGAACATGATCGACAA-3′ | This study |
| AlrKO2 | 5′-AAATTTGGTACCGGTCTGCATCGTCATAATCT-3′ | This study |
| AlrKO3 | 5′-AAATTTAAGCTTGGCAGGACAACAAGATTGAG-3′ | This study |
| AlrKO4 | 5′-AAATTTTCTAGAGCCCTCGACATCCATTGCTT-3′ | This study |
| 5alr2 | 5′-AAATTTCCATGGTGGGGCAGTACTACAACTTC-3′ | This study |
| 3alr2 | 5′-AAATTTAAGCTTGTAGGGGTCTTCCTTGCTCA-3′ | This study |