TABLE 1.
Bacterial strains, plasmids, and oligonucleotides
| Material | Relevant characteristics | Reference or Source |
|---|---|---|
| Strains | ||
| Mycobacterium rhodesiae JS60 | Grows on VC and ethene as sole carbon sources | 7 |
| Mycobacterium smegmatis mc2155 | Highly electrotransformable derivative of ATCC 607 | 40 |
| Escherichia coli JM109 | recA1 supE44 endA1 hsdR17 gyrA96 relA1 thi Δ(lac-proAB) F′[traD36 proAB+laclqlacZ ΔM15] | 58 |
| Plasmids | ||
| pK18 | Kmr, 2.7 kb, general purpose cloning vector | 34 |
| pGEM-T Easy | Apr, 3.0 kb, T/A cloning vector | Promega |
| pMV261 | Kmr, 4.5 kb, E. coli-Mycobacterium shuttle vector | 41 |
| pMV-CoM | Kmr, 5.8 kb, pMV261 carrying JS60 EaCoMT gene (etnE) | This study |
| Oligonucleotides (5′-3′) | ||
| CoM-F1 | ATGGTGGGGAACTACCCGAATCC | This study |
| CoM-R1 | ATGAGGCCGCAGTCGGTGGACA | This study |
| CoM-F1L | AACTACCCSAAYCCSCGCTGGTACGAC | This study |
| CoM-R2 | TCGTCGGCAGTTTCGGTGATCGTGCTCTT | This study |
| 60M-F2 | GGGGATATCAGTGATCGCCATACAGCAATTAAGGAG | This study |
| 60M-R2 | GGGAAGCTTTTGTGTCCGGACAGGCCGATAGTC | This study |
| RTM-F1 | AAGGAGTGCATCATGAAGGTTGGGG | This study |
| RTβ-R1 | TTGGCTGATACCGGGTCGAGGTTGAG | This study |