Table 1. Strains and plasmids used in this work.
| Strain or plasmid | Relevant features | Source or reference |
|---|---|---|
|
E. coli |
||
| DH5α | High-efficiency transformation | Bethesda Research Laboratories |
| SM10 | Conjugation proficient E. coli donor strain | [39] |
|
B. pertussis |
||
| Tohama I | Patient isolate | [40] |
| BP536 | Tohama I, StrR, NalR | [31] |
| BP953 | BP536, fhaB-lacZ, ptx-phoA | [17] |
| BP1056 | Pho-, Lac+, Hly- mutant of BP953 (bvgAD201N) | [17] |
| BP1286 | Pho+ mutant of BP1056 (bvgAD201N, Δ127–129) | This study |
| BP1318 | BP953 bvgAD201N, Lac+, Pho-, created by allelic exchange (pSS2423). Clean version of BP1056. | This study |
| BP1324 | BP953 bvgAD201N, Δ127–129, Lac+, Pho+, created by allelic exchange (pSS2429). Clean version of BP1286. | This study |
| BP1322 | BP953 bvgAΔ127–129, Lac+, Pho+, created by allelic exchange (pSS2427) | This study |
| QC3216 | (BP536 ΔbvgA::P2bvgS) | [9] |
|
Plasmids |
||
| pSS1827 | Helper plasmid for triparental mating, RP4 tra genes, AmpR |
[41] |
| pTTQ8 | Ptac expression vector | [34] |
| pSS1894 | Suicide vector, oriT, gen, amp, | [33] |
| pSS2000 | pSS1894::rpsL | This study |
| pSS1574 | pTTQ8::phoA | This study |
| pSS2072 | pSS1574::aph | This study |
| pSS2075 | Suicide vector, oriT, gen, amp, ‘phoA-aph | This study |
| pSS2197 | bvgA allelic retrieval vector | [18] |
| pSS2401 | Derived by allelic retrieval of bvgA from BP1286 using pSS2197 | This study |
| pSS2423 | pSS2401 derivative containing bvgAD201N | This study |
| pSS2427 | pSS2401 derivative containing bvgAΔ127–129 | This study |
| pSS2429 | pSS2401 derivative containing bvgAD201N, Δ127–129 | This study |
| pSS3967 | Ectopic luxCDABE promoter assay vector, Ampr Genr | [7,25] |
| pQC1114 | pSS3967::WT Pptx (-290 to +28 relative to Pptx +1) | This work |
| pQC1384 | pSS3967::PΔ1 (-290 to +28 relative to Pptx +1) | This work |
| pQC1385 | pSS3967::PΔ2 (-290 to +28 relative to Pptx +1) | This work |
| pQC1115 | pSS3967::PΔ3 (-290 to +28 relative to Pptx +1) | This work |
| pQC1386 | pSS3967::PΔ4 (-290 to +28 relative to Pptx +1) | This work |
| pQC1119 | pSS3967::PΔ5 (-290 to +28 relative to Pptx +1) | This work |
| pQC1383 | pSS3967::PΔ6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1116 | pSS3967::BS1-2-5-6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1120 | pSS3967::BS1-2-3-6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1117 | pSS3967::BS1-2-6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1468 | pSS3967::BS1-3-6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1584 | pSS3967::BS1-4-6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1585 | pSS3967::BS1-5-6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1469 | pSS3967::BS1-6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1488 | pSS3967::BS1-3 (-290 to +28 relative to Pptx +1) | This work |
| pQC1489 | pSS3967::BS3-6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1490 | pSS3967::S-BS6 (-290 to +28 relative to Pptx +1) | This work |
| pQC1467 | pSS3967::TTGACA (-290 to +28 relative to Pptx +1) | This work |
| pQC1487 | pSS3967::TTTTTT (-290 to +28 relative to Pptx +1) | This work |
| pQC1557 | pSS3967:: WT Pfha- (BS+30, -124 to +4)-lux in pSS3967 | [9] |
| pQC1552 | pSS3967::WT Pptx-BS+30 (-190 to +4 relative to Pptx +1) | This work |
| pQC2241 | Ectopic rfp promoter assay vector, Ampr Genr | This work |
| pQC2306 | pQC2241::WT Pfha-BS+30 (-125 to +4 relative to Pfha +1) | This work |
| pQC2307 | pQC2241:: WT Pptx-BS+30 (-190 to +4 relative to Pptx +1) | This work |
| pSS4162 | In situ luxCDABE promoter assay vector, Ampr Genr | [25] |
| pQC2623 | pSS4162:: WT Pfha (-970 to +4 relative to Pfha +1) | This work |
| pQC2624 | pSS4162::WT Pptx (-998 to +4 relative to Pptx +1) | This work |
| pQC2319 | In situ rfp promoter assay vector, Ampr Genr | [25] |
| pQC2626 | pQC2319:: WT Pfha (-970 to +4 relative to Pfha +1) | This work |
| pQC2627 | pQC2319:: WT Pptx (-998 to +4 relative to Pptx +1) | This work |
| pQC1883 | B. pertussis expression vector, Kanr | [9] |
| pSS4983 | pQC1883::bvgA, Kanr | [9] |
| pQC1894 | pQC1883::bvgAΔ127–129 | This work |