Table 1.
Oligonucleotides used for amplification and cloning of l,d-transpeptidase genes
| l,d-Transpeptidase (strain; accession no.) and amplicona | Oligonucleotideb |
|---|---|
| Mt2 (Rv2518c; CAA16014.1) | |
| C | 5′AACATATGGCCGATCTGCTGGTGCC (for) |
| D | 5′TTCTCGAGTTACGCCTTGGCGTTACCGG (rev) |
| Mt3 (Rv1433; CAB09251.1) | |
| A | 5′TATGCAGTCTTACGGGTTCGCCGT (for) |
| 5′GTTATTCCTGCACAATGACCGGGT (rev) | |
| B | 5′TGCAGTCTTACGGGTTCGCCGT (for) |
| 5′TCGAGTTATTCCTGCACAATGACCGGGT (rev) | |
| Mt4 (Rv0192; CAB09732.1) | |
| C | 5′CATATGCCACACTGGGCTGAAGAACG (for) |
| D | 5′CTCGAGTTAGATCTGCCAGTCCTGGGCACC (rev) |
| Mt5 (Rv0483; CAB00944.1) | |
| A | 5′TATGGCCGGCAAAGTGACCAAGCT (for) |
| 5′GTTACCCACCCGGTCCGTTAGTAG (rev) | |
| B | 5′TGGCCGGCAAAGTGACCAAGCTGG (for) |
| 5′TCGAGTTACCCACCCGGTCCGTTAGTAG (rev) |
a
For cloning, amplicons A and B were mixed, denatured, and renatured to generate heteroduplexes with cohesive ends.
b
Initiation and stop codons are in italic. For Mt2 and Mt4, oligonucleotides C and D contained NdeI and XhoI sites (underlined). for, forward; rev, reverse.