TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Antibiotic resistancea | Reference or source |
|---|---|---|---|
| F. tularensis subsp. holarctica strains | |||
| LVS | Live vaccine strain | 8 | |
| ST1101 | LVS derivative, ΔmglA | 75 | |
| ST1116 | LVS derivative, ΔclpX | This study | |
| ST1118 | LVS derivative, ΔclpP | This study | |
| ST1120 | LVS derivative, Δlon | This study | |
| ST1284 | LVS derivative, tig::Tn, transposon insertion between nucleotides 1104 and 1105 in tig | Kanr | 8 |
| ST1292 | LVS derivative, clpP::Tn, transposon insertion between nucleotides 255 and 256 in clpP | Kanr | 8 |
| ST1796 | ST1118 (pST1760); ΔclpP mutant complemented in trans with the wild-type LVS clpP driven by native tig promoter | Hygr | This study |
| ST1889 | ST1120 (pST1864); LVS Δlon mutant complemented in trans with wild-type LVS lon driven by native tig promoter | Hygr | This study |
| E. coli strains | |||
| DH5α | F− endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 hsdR17(rK− mK+) λ− | Biomed (Beijing, China) | |
| TOP10F′ | F′ [lacIq Tn10(Tetr)] mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80lacZΔM15 ΔlacX74 deoR nupG recA1 araD139 Δ(ara-leu)7697 galU galK rpsL(Strr) endA1 λ− | Invitrogen | |
| BL21 | B F− dcm ompT hsdS(rB− mB−) gal | Biomed | |
| Rosetta | B F− dcm ompT hsdS(rB− mB−) gal dcm λ (DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) | Biomed | |
| ER2566 | F− λ− fhuA2 [lon] ompT lacZ::T7 gene 1 gal sulA11 Δ(mcrC-mrr)114::IS10R(mcr-73::miniTn10-Tets)2 R(zgb-210::Tn10)(Tets) endA1 [dcm] | NEB | |
| Plasmids | |||
| pACYCDuet-1 | E. coli protein expression vector | Cmr | Novagen |
| pBS | pBluescript II (SK−) | Ampr | Stratagene |
| pBAD18 | E. coli protein expression vector | Ampr | 53 |
| pDL | Bacillus subtilis integration vector with bgaB of Bacillus stearothermophilus as a reporter gene | Cmr Ampr | Bacillus Genetic Stock Center |
| pEDL17 | Francisella-E. coli shuttle plasmid | Hygr | 57 |
| pET28a | E. coli protein expression vector | Kanr | Novagen |
| pGEX-2T | E. coli protein expression vector | Ampr | GE Healthcare Bio-Science |
| pMP590 | F. tularensis suicide vector | Kanr | 76 |
| pMP633 | Francisella-E. coli shuttle plasmid | Hygr | 76 |
| pST1010 | pBS::ΔclpP | Ampr | This study |
| pST1012 | pBS::ΔclpX | Ampr | This study |
| pST1100 | pBS::Δlon | Ampr | This study |
| pST1105 | pMP590::Δlon | Kanr | This study |
| pST1136 | pMP590::ΔclpP | Kanr | This study |
| pST1138 | pMP590::ΔclpX | Kanr | This study |
| pST1728 | pMP633::pTAC | Hygr | This study |
| pST1760 | pST1728::TIG-clpP | Hygr | This study |
| pST1864 | pST1728::pTIG-lon | Hygr | This study |
| pST1923 | pST1728::bgaB | Hygr | This study |
| pST1934 | pST1923::pTIG-bgaB | Hygr | This study |
| pST1936 | pST1923::pLON-bgaB | Hygr | This study |
| pST1938 | pST1923::pCLPB-bgaB | Hygr | This study |
| pST1940 | pST1923::pHUPB-bgaB | Hygr | This study |
| pST1956 | pST1923::pCLPP-bgaB | Hygr | This study |
| pST1958 | pST1923::pCLPX-bgaB | Hygr | This study |
| pST2700 | pGEX-2T derivate; a multiple-cloning site was added | Ampr | This study |
| pST3001 | pST2700::lon | Ampr | This study |
| pST3621 | pET28a derivate; an AscI site was added | Kanr | This study |
| pST3749 | pST2700::clpX | Ampr | This study |
| pST3750 | pST2700::clpP | Ampr | This study |
| pST4218 | pST3621::FTL1017 | Kanr | This study |
| pST4609 | pACYCDuet-1::FTL663 | Cmr | This study |
| pST4611 | pACYCDuet-1::FTL1957 | Cmr | This study |
| pST4635 | pBAD18::lon | Ampr | This study |
| pST4816 | pACYCDuet-1::FTL478 | Cmr | This study |
| pST4818 | pACYCDuet-1::FTL1017 | Cmr | This study |
| pST4819 | pACYCDuet-1::clpX | Cmr | This study |
| pST4861 | pEDL17::FTL663 | Hygr | This study |
| pST4863 | pEDL17::FTL1957 | Hygr | This study |
| pST4865 | pEDL17::FTL478 | Hygr | This study |
| pST4867 | pEDL17::FTL1017 | Hygr | This study |
| pST4869 | pEDL17::clpX | Hygr | This study |
| pST5007 | pACYCDuet-1::FTL578 | Cmr | This study |
| pST5009 | pACYCDuet-1::FTL1217 | Cmr | This study |
| pST5072 | pEDL17::FTL1217 | Hygr | This study |
| pST6927 | pACYCDuet-1::FTL40 | Cmr | This study |
| pST6928 | pACYCDuet-1::FTL398 | Cmr | This study |
| pST6930 | pACYCDuet-1::FTL687 | Cmr | This study |
| pST6931 | pACYCDuet-1::FTL891 | Cmr | This study |
| pST6935 | pACYCDuet-1::FTL1191 | Cmr | This study |
| pST6936 | pACYCDuet-1::FTL1228 | Cmr | This study |
| pST6937 | pACYCDuet-1::FTL1267 | Cmr | This study |
| pST6938 | pACYCDuet-1::FTL1358 | Cmr | This study |
| pST6939 | pACYCDuet-1::FTL1609 | Cmr | This study |
| pST6955 | pEDL17::FTL578 | Hygr | This study |
| pST6956 | pEDL17::FTL1228 | Hygr | This study |
| pST7043 | pACYCDuet-1::FTL87 | Cmr | This study |
| pST7044 | pACYCDuet-1::FTL593 | Cmr | This study |
| pST7045 | pACYCDuet-1::FTL595 | Cmr | This study |
| pST7046 | pACYCDuet-1::FTL599 | Cmr | This study |
| pST7048 | pACYCDuet-1::FTL1611 | Cmr | This study |
| pST7049 | pACYCDuet-1::FTL1724 | Cmr | This study |
a
Ampr, ampicillin resistance; Cmr, chloramphenicol resistance; Kanr, kanamycin resistance; Hygr, hygromycin resistance.