TABLE 4.
Primers used in this study
| Primer use and name | Amplified region | Approximate size (bp) | Sequence (5′–3′)a |
|---|---|---|---|
| qPCR | |||
| RTDIP2330_1 | hbpA | 152 | CTCTCGGGTGGAGAACAAAG |
| RTDIP2330_2 | CTGCCTTGGAGTTGAGGAAG | ||
| RTDIP2329_1 | dip2329 | 226 | CTGATCCGCCAACAAGGTAT |
| RTDIP2329_2 | TGTGGATTTGGCTGTGGTTA | ||
| RTDIP2331_1 | dip2331 | 234 | ACGAACCACTGGGTGTTCTC |
| RTDIP2331_2 | CGGAAAGGATTGTCTCGGTA | ||
| RTgyrB1 | gyrB | 166 | GGTCTGACCATTACGCTGGT |
| RTgyrB2 | TCTTCTCGCGTTTCTTTGGT | ||
| RTtox1 | tox | 224 | GAACAGGCGAAAGCGTTAAG |
| RTtox2 | TTTTTGATAGGGCCATGCTC | ||
| EMSA | |||
| P2330 Fwd1 biotin | PhbpA | 377 | Biotin-CCCCCCCGTTTCCTCCGGTGAAATAATTA |
| P2330 Rev1 biotin | Biotin-CCCCCCATTCAAGATTGATTCCTTTGAGGG | ||
| P2330 Fwd2 biotin | P3′ hbpA | 175 | Biotin- CCCCCCGGCCCCTAACTGACAAATTAG |
| P2330 Rev1 biotin | Biotin-CCCCCCATTCAAGATTGATTCCTTTGAGGG | ||
| P2330 Fwd1 biotin | P5′ hbpA | 205 | Biotin-CCCCCCCGTTTCCTCCGGTGAAATAATTA |
| P2330 Rev2 biotin | Biotin-CCCCCCGCCTATTTTTCCTATATGCAAAGG | ||
| Ptox Fwd biotin | tox | 232 | Biotin-CCCCCCCTCATTGAGGAGTAGGTCCC |
| Ptox Rev biotin | Biotin-CCCCCCCATGGGCTGAAGGTGGGG | ||
| Promoter fusion | |||
| P2330 Fwd | hbpA | 377 | GCACGTCGACCGTTTCCTCCGGTGAAATAATTA |
| P2330 Rev | GATCGGATCCATTCAAGATTGATTCCTTTGAGGG | ||
| Complement clone | |||
| P2330 Fwd | hbpA pKN2.6Z | 1,436 | GCACGTCGACCGTTTCCTCCGGTGAAATAATTA |
| 2330 compl Rev | GACTGGATCCTGGGGCTTAACGCACGAAC | ||
| Expression clone | |||
| 2330 streptag Fwd | hbpA pET24(a)+ | 873 | GATCGCTAGCTGGAGCCACCCGCAGTTCGAAAAG |
| GGTGCAGCAGAAGAAGTAAAAAATGCCG | |||
| 2330 Rev | GATGAAGCTTTTATGCCTTGGAGTTGAGGAAGC | ||
| Deletion mutants | |||
| 2330 5′ KO Fwd | Up hbpA | 744 | GATGGGATCCACAAGGTATTCACGTTCTCCG |
| 2330 5′ KO Rev | GGTGGTCAGGATATTCAAGATTG | ||
| 2330 3′ KO Fwd | Down hbpA | 762 | CAATCTTGAATATCCTGACCACCCATTTGTTC |
| GTGCGTTAAGCC | |||
| 2330 3′ KO Rev | GTTCGTCGACGTTGACTAGGAGGCCTTCG |
a
Restriction sites are underlined, and the Strep-tag sequence is indicated in bold type.