Table 1.
Strain and plasmid list.
| Strain/plasmid | Serotype and relevant genotype | Resistance* | Source or reference |
|---|---|---|---|
| Streptococcus pneumoniae | |||
| SP257 (D39) | 2 | None | NCTC7466 |
| PN111 | D39Δcps | Kmr | (30, 31) |
| PN282 | D39ΔcpsΔadcAII (spd_0888) | Kmr, Ermr | This work |
| PN279 | D39ΔcpsΔdacB (spd_0549) | Kmr, Ermr | (9) |
| PN253 | D39ΔcpsΔetrx1 (spd_0572) | Kmr, Ermr | (10) |
| PN281 | D39ΔcpsΔetrx2 (spd_0886) | Kmr, Ermr | (10) |
| PN238 | D39ΔcpsΔmetQ (spd_0151) | Kmr, Ermr | This work |
| PN732 | D39ΔcpsΔppmA (spd_0868) | Kmr, Trmr | This work |
| PN280 | D39ΔcpsΔpnrA (spd_0739) | Kmr, Ermr | This work |
| PN301 | D39ΔcpsΔpsaA (spd_1463) | Kmr, Ermr | This work |
| PN733 | D39ΔcpsΔslrA (spd_0672) | Kmr, Ermr | This work |
| PN241 | D39ΔcpsΔspd_0179 | Kmr, Ermr | This work |
| PN312 | D39ΔcpsΔspd_0792 | Kmr, Ermr | This work |
| PN735 | D39ΔcpsΔpspA (spd_0126) | Kmr, Ermr | This work |
| PN278 | D39ΔadcAII (spd_0888) | Ermr | This work |
| PN275 | D39ΔdacB (spd_0549) | Ermr | (9) |
| PN246 | D39Δetrx1 (spd_0572) | Ermr | (10) |
| PN277 | D39Δetrx2 (spd_0886) | Ermr | (10) |
| PN311 | D39ΔmetQ (spd_0151) | Ermr | This work |
| PN093 | D39ΔppmA (spd_0868) | Trmr | (32) |
| PN276 | D39ΔpnrA (spd_0739) | Ermr | This work |
| PN172 | D39ΔpsaA (spd_1463) | Ermr | (13) |
| PN095 | D39ΔslrA (spd_0672) | Ermr | (32) |
| PN243 | D39Δspd_0179 | Ermr | This work |
| PN251 | D39Δspd_0792 | Ermr | This work |
| PN031 | D39ΔpspA (spd_0126) | Ermr | (33) |
| Escherichia coli | |||
| DH5α | Δ(lac)U169, endA1, gyrA46, hsdR17, Φ80Δ(lacZ)M15, recA1, relA1, supE44, thi-1 | None | Bethesda Research Labs, Gaithersburg, U.S. |
| BL21(DE3) | E. coli B, F- dcm ompT hsdS gal λ(DE3), T7 polymerase gene under control of the lacUV5 promoter | None | Novagen, Merck KGaA, Darmstadt, Germany |
| Plasmids | |||
| pGEM®-T easy | TA cloning vector for PCR products | Apr | Madison, U.S. |
| p89 | pCR2.1Topo with erythromycin (ermB) cassette | Apr, Kmr, Ermr | (34) |
| p873 | pGXT with capsule locus replaced by aphA3 gene resistance cassette, flanking genes dex ,and aliA | Apr, Kmr | (10) |
| p572 | pGEM-T derivative with sp_1002 (adcAII) 5′ and 3′ flanking region for mutagenesis | Apr | This work |
| p598 | pGEM-T derivative with sp_1002 (adcAII) interrupted by ermB gene resistance cassette | Apr, Ermr | This work |
| p559 | pGEM-T derivative with sp_0149 (metQ) 5′ and 3′ flanking region for mutagenesis | Apr | (28) |
| p563 | pGEM-T derivative with sp_0149 (metQ) interrupted by ermB gene resistance cassette | Apr, Ermr | (28) |
| p576 | pGEM-T derivative with sp_0845 (pnrA) 5′ and 3′ flanking region for mutagenesis | Apr | This work |
| p646 | pGEM-T derivative with sp_0845 (pnrA) interrupted by ermB gene resistance cassette | Apr, Ermr | This work |
| p560 | pGEM-T derivative with sp_0191 (spd_0179) 5′ and 3′ flanking region for mutagenesis | Apr | This work |
| p565 | pGEM-T derivative with sp_0191 (spd_0179) interrupted by ermB gene resistance cassette | Apr, Ermr | This work |
| p573 | pGEM-T derivative with sp_0899 (spd_0792) 5′ and 3′ flanking region for mutagenesis | Apr | This work |
| p577 | pGEM-T derivative with sp_0899 (spd_0792) interrupted by ermB gene resistance cassette | Apr, Ermr | This work |
| pTP1 | pET28 expression vector, N-terminal His-tag, TEV protease cleavage site, induction by IPTG | Kmr, Ermr | (10) |
| p648 | pTP1 with TIGR4 sp_1002 (adcAII) for protein production and mice immunisation | Kmr | This work |
| p652 | pTP1 with TIGR4 sp_0629 (dacB) for protein production and mice immunisation | Kmr | (9) |
| p629 | pTP1 with TIGR4 sp_0659 (etrx1) for protein production and mice immunisation | Kmr | (10) |
| p651 | pTP1 with TIGR4 sp_1000 (etrx2) for protein production and mice immunisation | Kmr | (10) |
| P732 | pTP1 with TIGR4 sp_0149 (metQ) for protein production and mice immunisation | Kmr | This work |
| p264 | pET11a with R6 spr0884 (ppmA) for protein production and mice immunisation | Kmr | (32) |
| p649 | pTP1 with TIGR4 sp_0845 (pnrA) for protein production and mice immunisation | Kmr | This work |
| p653 | pTP1 with TIGR4 sp_1650 (psaA) for protein production and mice immunisation | Kmr | This work |
| p263 | pET11a with R6 spr0679 (slrA) for protein production and mice immunisation | Kmr | (32) |
| p628 | pTP1 with TIGR4 sp_0191 (spd_0179) for protein production and mice immunisation | Kmr | This work |
| p631 | pTP1 with TIGR4 sp_0899 (spd_0792) for protein production and mice immunisation | Kmr | This work |
| p105 | pQE30 with PspA (aa 32-289) without choline-binding domain | Apr | (33, 68) |
*
Kmr, Kanamycin; Ermr, Erythromycin; Apr, Ampicillin; Trmr, Trimethoprim.