Table 1.
Bacterial strains, phages, plasmids and oligonucleotides used in this study.
| Name | Description | Source or Reference |
|---|---|---|
| Bacteria | ||
| Mycobacterium smegmatis mc2155 | High-transformation-efficiency mutant of M. smegmatis ATCC 607 | [26] |
| Bacteriophages | ||
| Ms6wt | Temperate bacteriophage from M. smegmatis | [27] |
| Ms6ΔlysB | 996 bp in-frame deletion of the Ms6 lysB gene | This study |
| Plasmids | ||
| pJV53 pAG1 |
Derivative of pLAM12 with Che9c 60 and 61 under control of the acetamidase promoter; Kanr lysB gene cloned into pVVAP |
[28] This study |
| pVVAP | Mycobacteria shuttle vector carrying the acetamidase promoter; Kanr | [29] |
| Oligonucleotides | Sequence 5’-3’ a | |
| PrΔlysB | CTCGGCGGAAAAACCCTCCTCGTGGACGCGGTAGCAGAACTGTTGGGCCACTGATAGGAGGCACCCATGCTGACACGTTCATTCTGGATCGACGCCGCCGAGCG | Ms6ΔlysB |
| PrExtΔlysBFw | CGAGATCCTGCGGCAACTGCGCGGATACAACCTCACTGGCTGGCCGCAGCTCGGCGGAAAAACCCTCGTGGACG | Extend Pr∆lysB |
| PrExtΔlysBRv | CCCCGGCGCCGAGGGTGGCGATCGCGGTTTGGGCGAATGTGCGTATGGCACGCTCGGCGGCGTCGATCCAGAATG | Extend Pr∆lysB |
| PrlysBFw | GCGGATCCATGAGCAGAACTGTTGGGCC | Includes BamHI site to clone in pVVAP |
| PrlysBRv | GGAAGCTTTGTGCGTAGGTAGTCGATG | Includes HindIII site to clone in pVVAP |
| DADA ΔlysB PCRFw | GCGCTAGCAGAACTGTTGGGCCACTGATAG | Ms6ΔlysB |
| DADA Ms6-PCRRv | CGTCTCGTACTGCACGTACCGGTTCTTC | Ms6ΔlysB |
a Restriction sites are underlined.