TABLE 1.
Oligonucleotides used for identification of mycobacterial DNA and cloning and screening of dehalogenase genes
| Primer | Target gene(s) | Sequence (5′-3′)a | Position | Source or reference |
|---|---|---|---|---|
| UNB51 | Bacterial 16S rRNA | GAGTTTGATCCTGGCTCA | 8-27 | 21 |
| UNB800 | Bacterial 16S rRNA | GGACTACCAGGGTATCTAAT | 787-806 | 21 |
| YNP9 | dnaJ (heat shock protein) | GGGTGACGCGGCATGGCCCA | 13-32 | 28 |
| YNP10 | dnaJ (heat shock protein) | CGGGTTTCGTCGTACTCCTT | 229-248 | 28 |
| P90 | Insertion sequence IS900 | GAAGGGTGTTCGGGGCCGTCGCTTAGG | 15-41 | 13 |
| P91 | Insertion sequence IS900 | GGCGTTGAGGTCGATCGCCCACGTGAC | 401-427 | 13 |
| DMBAf | dmbA | GCCGAATTCTAAGGAGGAATATCGATGACAGCATTCGGC | Cloning primerb | This study |
| DMBAr | dmbA | GCCAAGCTTTCAGTGATGGTGATGGTGATGGACGCCGGCGGCCGCCGACCG | Cloning primer | This study |
| DMBBf1 | dmbB | GCCGAATTCTAAGGAGGAATATCGATGGATGGATGTCCTACGC | Cloning primer | This study |
| DMBBr1 | dmbB | GCCAAGCTTTTAGTGATGGTGATGGTGATGCGTTGCCTGCTGCCAGGA | Cloning primer | This study |
| DMBBf2 | dmbB | GCCGGATCCTATGGATGTCCTACGCACC | Cloning primer | This study |
| DMBBr2 | dmbB | GCNCTGCAGTTAGTGATGGTGATGGTGATGCGTTGCCTGCTGGGAG | Cloning primer | This study |
| RV1f | dmbA | GCTATAGCTATGGCGAGCAA | 239-258 | This study |
| RV5f | dmbA | GTTGTCGTGGCCACGAAAC | 615-633 | This study |
| RV7r | dmbA | GCTGTCCTCCTGAACGAAAT | 818-837 | This study |
| RV8f | dmbA | GGGACCCGACCGCTATAGC | 228-246 | This study |
| TBC4r | dmbB | CGGGGAAAGGTGCATCGTAG | 585-604 | This study |
| TBC5f | dhmA | TTCGAAAACCTGGAGGACTAC | 31-51 | This study |
| TBC6r | dmbB, dhmA | GAAAGCCGTTGGCCACCACC | 429-448 | This study |
| TBC7f | dmbB, dhmA | GCACGGCGAGCCCACCTGGAG | 156-176 | This study |
a
Restriction sites are underlined.
b
Cloning primer, primers designed in reference to flanking regions of cloned genes. To the 5′ end of each primer specific restriction sequences were added.