Table 1.
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) |
For cloning, amplicons A and B were mixed, denatured, and renatured to generate heteroduplexes with cohesive ends.
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.