TABLE 2.
Primers used in this study
| Primer | DNA sequence (5′ to 3′)a | Purpose |
|---|---|---|
| pBBRdmt02-F | GGGGTACCCATGTTGTCGGGGTGAAGGAAAGTG (KpnI) | Amplification of a fragment containing the methyltransferase gene from scaffold 02 for functional study in strain DC-6 |
| pBBRdmt02-R | GCTCTAGATGTCCTGGGCCGCAAGGGCGAGTT (XbaI) | |
| pBBRdmt02c-F | CGGGGTACCCAATGGCGGCCGCCCTGATGCTC (KpnI) | Amplification of a fragment containing the methyltransferase gene cluster from scaffold 02 for functional study in strain DC-6 |
| pBBRdmt02c-R | CCGGAATTCTGGCTTCGGCATCGCCACCGAATGC (EcoR I) | |
| pBBRdmt66-F | CGGAATTCGGGTCAGGGCGCGGCCGCTAGG (EcoR I) | Amplification of a fragment containing the methyltransferase gene from scaffold 66 for functional study in strain DC-6 |
| pBBRdmt66-R | GCTCTAGAGCACGGCGAGAATAACCCCGAAGCA (XbaI) | |
| pBBRdmt66c-F | CTAGTCTAGACGTTGCAGCTCATCAACACCCACGA (XbaI) | Amplification of a fragment containing the methyltransferase gene cluster from scaffold 66 for functional study in strain DC-6 |
| pBBRdmt66c-R | CCCAAGCTTTCAAGTCCGCCATCAGGAGCGAA (HindIII) | |
| pET-dmt02-F | GGAATTCCATATGAAGAGCCCATTGATGAAATATCGG (NdeI) | Amplification of the methyltransferase gene from scaffold 02 for expression in Escherichia coli |
| pET-dmt02-R | CCGCTCGAGTCAGTCTTTCTTGATGACCTTCTCG (XhoI) | |
| pET-dmt66-F | GGAATTCCATATGGTGCGGTCGGTTCAGGA (NdeI) | Amplification of the methyltransferase gene from scaffold 66 for expression in Escherichia coli |
| pET-dmt66-R | CCGCTCGAGGAGCGTCGCGCGGACCCGGC(XhoI) | |
| RTdmt-F | GCCTTTCATTCCTGCCGAGTTCA | Amplification of a 525-bp fragment of dmt by RT-PCR |
| RTdmt-R | ATGCCATGCCGCAGGATGTGAACGC | |
| RTmetF-F | CGGCGCTCATCGTCACGCAATT | Amplification of a 301-bp fragment of metF by RT-PCR |
| RTmetF-R | AAAGGGTAGAAGTGCAGGCGGACAT | |
| RTpurU-F | TTGACCATTGCCTGGGTGACCTGCT | Amplification of a 540-bp fragment of purU by RT-PCR |
| RTpurU-R | TTCAGGAAGACCCGGCCCTCCAGAT | |
| RTdhc-F | CATAAAGCCTGCGAAGGACGTGGA | Amplification of a 166-bp fragment of dhc by RT-PCR |
| RTdhc-R | TCGACTTTCCGATCACCACAGCG | |
| RTdmt-metF-F | TTTACACCAGCGACGCAATGAA | Amplification of a 1,167-bp fragment of dmt-metF-spanning region |
| RTdmt-metF-R | CTGGCAATCAACTGCATGCTATCGG | |
| RTmetF-purU-F | GTCCGCATCCGTGCTGAGCAAATAT | Amplification of a 604-bp fragment of metF-purU-spanning region |
| RTmetF-purU-R | GAACGGGACGCCACGGAGCGATGAG | |
| RTpurU-dhc-F | ATCGCCCAGGATACCGAGGTCGT | Amplification of a 663-bp fragment of purU-dhc-spanning region |
| RTpurU-dhc-R | TCGACTTTCCGATCACCACAGCG | |
| RT-dmt-F | GCCTTTCATTCCTGCCGAGTTCA | Amplification of a 161-bp fragment of dmt by RT-qPCR |
| RT-dmt-R | TGGAGAAGGTGGACGGCGACAGATA | |
| RT-metF-F | GTCCGCATCCGTGCTGAGCAAATAT | Amplification of a 138-bp fragment of metF by RT-qPCR |
| RT-metF-R | AAAGGGTAGAAGTGCAGGCGGACAT | |
| RT-purU-F | TTGACCATTGCCTGGGTGACCTGCT | Amplification of a 128-bp fragment of purU by RT-qPCR |
| RT-purU-R | GAACGGGACGCCACGGAGCGATGAG | |
| RT-dhc-F | CATAAAGCCTGCGAAGGACGTGGA | Amplification of a 167-bp fragment of dhc by RT-qPCR |
| RT-dhc-R | TCGACTTTCCGATCACCACAGCG | |
| RT-16S-F | GGCGACGATCCATAGCTGGTCTGAG | Amplification of a 141-bp fragment of 16S rRNA by RT-qPCR |
| RT-16S-R | TTCATCACTCACGCGGCATTGCTG |
a
Restriction sites are underlined.