Table 1.
Strains and plasmids used in this work
| Strain or plasmid | Derived, relevant genotype, or characteristica | Source or reference |
|---|---|---|
| Escherichia coli | ||
| χ6212 | ΔasdA4 | 43 |
| χ7213 | F−supE42 λ− T3rthi-1 thr-1 leuB6 supE44 tonA21 fhuA21 lacY1 recA1 RP4 2-Tc::Mu (λpir) ΔasdA4 Δ(zhf-2::Tn10) | 22 |
| LMG194 | F− ΔlacX74 galE thi rpsL ΔphoA(PvuII) Δara714 leu::Tn10 | Invitrogen |
| Salmonella enterica serovar Typhimurium | ||
| χ9879 | ΔasdA33 ΔPphoPQ176::TT araC PBADphoΔPcrp527::TT araC PBADcrp ΔaraBAD23 | 42 |
| χ11021 | ΔPmurA25::TT araC PBADmurA ΔasdA27::TT araC PBADc2ΔaraBAD23 Δ(gmd-fcl)-26 Δpmi-2426ΔrelA198::TT araC PBADlacI | This study |
| Mycobacterium | ||
| M. tuberculosis H37Rv | ATCC 25618 | |
| M. bovis BCG | Pasteur | ATCC 35734 |
| Plasmids | ||
| Suicide vectors | ||
| pYA3546 | pDMS197 Δpmi-2426 | 25 |
| pYA3629 | pMEG-375 Δ(gmd-fcl)-26 | 25 |
| pYA4064 | pRE112 ΔrelA198::TT araC PBADlacI | 34 |
| pYA4138 | pRE112 ΔasdA27::TT araC PBADc2 | 34 |
| pYA4280 | pRE112 Δ(araC PBAD)-5::PRaraBAD | 23 |
| pYA4343 | pRE112 ΔPphoPQ176::TT araC PBADphoPQ | 22 |
| pYA4686 | pRE112 ΔPmurA25::TT araC PBADmurA | Laboratory stock |
| Asd+ plasmids | ||
| pYA3342 | asdA+ vaccine vector, pBR ori | 43 |
| pYA3620 | asdA+ expression vector, Ptrc pBR ori blaSS-blaCT-T2SS signal sequence based | 44 |
| pYA3941 | pYA3620 blaSS-fbpA294 (optimized)-blaCT | This study |
| Asd+/MurA+ lysis plasmids | ||
| pYA3681 | asdA+murA+ expression vector, Ptrc pBR ori araC* PBADSD-GTG-asdA SD-GTG-murA P22 PR antisense mRNA | 46 |
| pYA4890 | pYA3681 sopENt80-esxA-esxA-esxB/(SD) blaSS-fbpA294-blaCT pBR ori | This study |
| pYA4893 | pYA3681 ompCSS-esxA-esxA-esxB/(SD) blaSS-fbpA294-blaCT pBR ori | This study |
| pYA4589 | asdA+murA+ expression vector, Ptrc p15A ori araC* PBADSD-GTG-asdA SD-GTG-murA P22 PR antisense mRNA | This study |
| pYA4891 | pYA4589 sopENt80-esxA-esxA-esxB/(SD) blaSS-fbpA294-blaCT p15A ori | This study |
| pYA4892 | pYA4595 sopENt80-esxA-esxA-esxB/(SD) blaSS-fbpA294-blaCT pSC101 ori | This study |
| pYA4894 | pYA4589 ompCSS-esxA-esxA-esxB/(SD) blaSS-fbpA294-blaCT p15A ori | This study |
| pYA4595 | asdA+murA+ expression vector, Ptrc pSC101 ori araC* PBADSD-GTG-asdA SD-GTG-murA P22 PR antisense mRNA | This study |
| pBK-CMV and its derivatives | ||
| pBK-CMV | Kmr, cloning vector | Stratagene |
| pYA3817 | pBK-CMV fbpA | This study |
| pYA3932 | pBK-CMV fbpA optimized | This study |
| Plasmids expressing 6×His-tagged recombinant proteins | ||
| pYA3814 | pBAD/His(B), 6×His-Ag85A | This study |
| pMRLB7 | Ampr, pET23+ ESAT-6–6×His | NIH-TB vaccine |
a
In the descriptions of the genotypes, TT is transcription terminator, P stands for promoter, and the subscripted number refers to a composite deletion and insertion of the indicated gene. Ampr, ampicillin resistance; Kmr, kanamycin resistance.