TABLE 3.
Oligonucleotides employed in this study
| Primer | Functiona | Sequences of oligonucleotide primer partnersb |
|---|---|---|
| Bup A and B | Amplification of BldB upstream region from the S. coelicolor chromosome | 5′-CAGGCGGCATATGAACGATCCCCGACAACCTTAC |
| 5′-GTCGAGAAGCTTTCGGGTGAGGCGCTG | ||
| Bdown A and B | Amplification of BldB downstream region from the S. coelicolor chromosome | 5′-CTCGACGAGCTCTGGCAGGGCACGCGC |
| 5′-GCGACGGGGCATATGTCGCCGAGTGAC | ||
| Bcomp A and B | Amplification of BldB and its promoter from the S. coelicolor chromosome | 5′-GCGACGCGAATTCGCTACGTCGCACCG |
| 5′-GTGCTCATCTCGGGTACGGGAGTGGTCCTGC | ||
| HisB A and B | Amplification of His-tagged bldB from pBB801 | 5′-GGAGGGTACCAATGAGAGGATCGCAT |
| 5′-TCGAGGTACCCCCTCGGCGACGACGCC | ||
| NHB | Amplification of non-His-tagged bldB from pBB801 | 5′-ATCAGGTACCAATGGCCCAGGTGCCG |
| pT18seq | Sequencing plasmid pT18 | 5′-GACCATGATTACGCCAAGCG |
| pT25seq | Sequencing plasmid pT25 | 5′-GATTCGGTGACCGATTACCTG |
| NT10 A and B | BldB aa 1-10 deletion in pT25BBH | 5′-GTGATGGTGATGGTGATGCGATC |
| 5′-GCCCGCAAGGAGCGCGAGC | ||
| NT20 A and B | BldB aa 11-20 deletion in pT25BBH | 5′-TTTGACGTCCTCGTCCGGCAC |
| 5′-TACGCGCTCGACATCTCGGGTG | ||
| NTVar | BldB aa 1-47 deletion in pT25BBH | 5′-CTGCCCGACGGAGCCGTGGCCATG |
| CT10 A and B | BldB aa 89-98 deletion in pT25BBH | 5′-ACCTGGAGCCGTAGCCGGGCGAC |
| 5′-GTCGCCCGGCTACGGCTCCAGGT | ||
| CT20 A and B | BldB aa 79-98 deletion in pT25BBH | 5′-TCGTCCTGGGTTAGCGGGACG |
| 5′-CGTCCCGCTAACCCAGGACGA | ||
| Framedel A and B | BldB aa 72-98 deletion in pT25BBH | 5′-GAGGCGGAGTGACGGGCTTTC |
| 5′-GAAAGCCCGTCACTCCGCCTC | ||
| T21L 1 and 2 | T21L mutagenesis in pT18BBH | 5′-CGAGCTGCTCGCGCTCGAC |
| 5′-GTCGAGCGCGAGCAGCTCG | ||
| T21C 1 and 2 | T21C mutagenesis in pT18BBH | 5′-CGAGCTGTGCGCGCTCGAC |
| 5′-GTCGAGCGCGCACAGCTCG |
a
aa, amino acids.
b
Nonhomologous regions and introduced restriction sites are in boldface type.