Table 2.
Primers used in this study
| Primer name | Sequence | Description |
|---|---|---|
| (in 5′ to 3′ direction) | ||
| Rev[VC1917]-NotI | GCGGCCGCGAGCTCTAGAGGTTTCTTAG | For inverse PCR leading to plasmids: |
| pBR-[−700]comEA, | ||
| pBR-[−500]comEA, | ||
| pBR-[−300]comEA, | ||
| pBR-[−100]comEA, | ||
| pBR-[−40]comEA | ||
| Fwd[VC1917]-700 | AGAGCTCGCGGCCGCAGGTGTTAACCACTCCTGCGGTAC | Inverse PCR to generate |
| pBR-[−700]comEA | ||
| Fwd[VC1917]-500 | AGAGCTCGCGGCCGCCAACAAGCACTTGAACTGGGTAAC | Inverse PCR to generate |
| pBR-[−500]comEA | ||
| Fwd[VC1917]-300 | AGAGCTCGCGGCCGCTATCGTTGTGATTGAGTTGAGC | Inverse PCR to generate |
| pBR-[−300]comEA | ||
| VC1917-134-NotI | GCGGCGGCCGCATTCTTAGTGTAATTGATATG | PCR to generate |
| pBR-TET_MCS after | ATCATGCGCACCCGTGGCCAGGACCC | pBR[−134]comEA, |
| Fwd[VC1917]-100 | AGAGCTCGCGGCCGCGGGCTACAGCAGTAGCCCGTTC | Inverse PCR to generate |
| pBR[−100]comEA | ||
| Fwd[VC1917]-40 | AGAGCTCGCGGCCGCCGCTATCATAAGCCCTCAACAAC | Inverse PCR to generate |
| pBR[−40]comEA | ||
| gyrA-157-fwd | AATGTGCTGGGCAACGACTG | qRT-PCR for gyrA transcription [10] |
| gyrA_332_bwd | GAGCCAAAGTTACCTTGGCC | |
| comEA_50_fwd | CGACATTACCGTTACTGGCC | qRT-PCR for comEA transcription [10] |
| comEA_224_bwd | CCGTTGGCTTCTCGATAATCG | |
| comEC_1029_fwd | GGTCGCGATTGTTCTCTACC | qRT-PCR for comEC transcription [10] |
| comEC_1186_bwd | CCAAATTGTACAGAACTGCCG | |
| VC0396_188_fwd | GCCTGATTCGCCAGCAATTG | qRT-PCR for qstR transcription [22] |
| VC0396_356_bwd | CCAAGACCGTGGGCAATAAAG | |
| hapR-230-fwd | CCAACTTCTTGACCGATCAC | qRT-PCR for hapR transcription [22] |
| hapR-399-bwd | GGTGGAAACAAACAGTGGCC | |
| hapA_175_fwd | ACGGTACAGTTGCCGAATGG | qRT-PCR for hapA transcription [22] |
| hapA_358_bwd | GCTGGCTTTCAATGTCAGGG | |
| comEA_284_rev | CGCACTGTCGCTTCACCAATCC | 5′RACE: synthesis of first strand cDNA of comEA |
| comEA_217_rev | CTTCTCGATAATCGACAATGGCCTGAGC | 5′RACE: PCR amplification of Poly(A) cDNA comEA |
| oligo dT-Anchor primer (Roche) | GACCACGCGTATCGATGTCGAC | |
| F-EcoRI_Anchor_P | CCAAGAATTCGACCACGCGTATCGATGTCGAC | 5′RACE: PCR fragment of Poly(A) cDNA comEA cloned into plasmid pBR-Tet_MCSI |
| R-BamHI_comEA_217 | CCAAGGATCCCTTCTCGATAATCGACAATGGCCTGAGC | |
| T7RNA-pol-750-down | GCTGAGGCTATCGCAACCCGTGC | DNA uptake assay: amplification of donor DNA; primer specific for E. coli BL21(DE3); [9,13] |
| T7 RNAP-end-bw | TTACGCGAACGCGAAGTCCGACTCTAAG | |
| lacZ-missing-fw | GCCGACTTTCCAATGATCCACAATGGG | DNA uptake assay: amplification of acceptor DNA; primer specific for V. cholerae A1552 and lacZ+ derivatives of it; [9,13] |
| lacZ-missing-bw | CCCTCGCTATCCCATTTGGAAATGCC |