TABLE 4 .
Key bacterial strains and plasmids used in this study
| Bacterial strain or plasmid |
Relevant characteristic(s) | Reference or source |
|---|---|---|
| E. coli strains | ||
| DH5α | F− φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) phoA supE44 λ− thi-1 gyrA96 relA1; used for general cloning |
51 |
| BL21(DE3) | F− ompT hsdSB(rB− mB−) gal dcm (DE3); used for protein overproduction | Novagen |
| χ6212 | F− λ− φ80 Δ(lacZYA-argF) endA1 recA1 hsdR17 deoR thi-1 glnV44 gyrA96 relA1 ΔasdA4 | 52 |
| χ7213 | thi-1 thr-1 leuB6 glnV44 fhuA21 lacY1 recA1 RP4-2-Tc::Mu(λpir) ΔasdA4 Δ(zhf-2::Tn10) | 46 |
| S. Typhimurium strains | ||
| χ3761 | Wild-type UK-1 | 45 |
| χ9088 | Δpmi-2426 Δ(gmd-fcl)-26 ΔPfur33::TT araC PBAD fur ΔasdA33 | 22 |
| χ11467 | Δ(agfC-agfG)-999 ΔsafABCD31 ΔstcABCD36 | χ11466 |
| χ11483 | ΔsafABCD31 ΔstiABCH1225 Δ(agfC-agfG)-999 | χ11468 |
| χ11484 | ΔsafABCD31 ΔstiABCH1225 Δ(agfC-agfG)-999 ΔstcABCD36 | χ11483 |
| χ11505 | ΔsafABCD31 ΔstiABCH1225 ΔstcABCD36 | χ11468 |
| χ11507 | Δ(agfC-agfG)-999 ΔstiABCH1225 ΔstcABCD36 | χ11506 |
| χ11595 | Δpmi-2426 Δ(gmd-fcl)-26 ΔPfur33::TT araC PBAD
fur ΔasdA33 ΔsafABCD31 Δ(agfC-agfG)-999 ΔstcABCD36 ΔPstiA52::PmurA stiA52 |
χ11594 |
| χ11599 | ΔsafABCD31 ΔstiABCH1225 ΔstcABCD36 Δ(agfC-agfG)-999 | χ11505 |
| χ11606 | Δpmi-2426 Δ(gmd-fcl)-26 ΔPfur33::TT araC PBAD
fur ΔasdA33 ΔsafABCD31 ΔstiABCH1225 Δ(agfC-agfG)-999 ΔstcABCD36 |
χ11597 |
| χ11850 | Δpmi-2426 Δ(gmd-fcl)-26 ΔPfur33::TT araC PBAD
fur ΔasdA33 ΔstiABCH1225 Δ(agfC-agfG)-999 ΔstcABCD36 ΔPsafA55::PmurA safA55 |
χ11594 |
| χ11851 | Δpmi-2426 Δ(gmd-fcl)-26 ΔPfur33::TT araC PBAD
fur ΔasdA33 ΔsafABCD31 ΔstiABCH1225 Δ(agfC-agfG)-999 ΔPstcA53::PmurA stcA53 |
χ11597 |
| χ12038 | Δpmi-2426 Δ(gmd-fcl)-26 ΔPfur33::TT araC PBAD
fur ΔasdA33 ΔsafABCD31 ΔstiABCH1225 ΔstcABCD36 agfD812 |
χ11562 |
| S. pneumoniae strain WU2 | Wild-type; virulent; encapsulated type 3 | 41 |
| Plasmids | ||
| pSG3 | IVET vector; promoterless aph lacZ mobRP4; R6K ori; Apr Gmr | 43 |
| Plasmids used for production of recombinant proteins |
||
| pET28b | Expression vector; T7 promoter 6xHis lacI; f1 pBR ori; Kmr | Novagen |
| pYA4085 | pET30a derivative for overproduction of rPspA | 49 |
| pYA4088 | pYA3493 derivative for production of rPspA (amino acids 3 to 285) fused to β-lactamase signal sequence |
25 |