Table 2.
Bacterial strains and plasmids used in this study.
| Strain | Genotype/Description | Reference |
|---|---|---|
| Shigella flexneri | ||
| 2457T | Wild-type Shigella flexneri 2a 2457T | Formal et al., 1958 |
| BS103 | Virulence plasmid-cured derivative of 2457T | Maurelli et al., 1984 |
| BS611 | 2457T/ΔicsA | Clark and Maurelli, 2007 |
| BS652 | 2457T/Δspa47 (spa47::aadA); SmRSpecR | Zurawski et al., 2006 |
| BS766 | 2457T transformed with pKM208; AmpR | Zurawski et al., 2006 |
| BS771 | 2457T/ΔospF (ospF::cat); CmR | Zurawski et al., 2006 |
| BS795 | 2457T/ΔospB (ospB::cat); CmR | This study |
| BS802 | BS771 transformed with pBAD24::ospF | Zurawski et al., 2006 |
| BS814 | 2457T/ΔospF; unmarked | Zurawski et al., 2006 |
| BS816 | 2457T/ΔospC1; unmarked | Zurawski et al., 2006 |
| BS818 | 2457T/ΔospB; unmarked | This study |
| BS820 | BS818 transformed with pDZ8; CmR | This study |
| BS821 | 2457T/ΔospZ; unmarked | Zurawski et al., 2008 |
| BS823 | BS652 transformed with pDZ8; CmR | This study |
| BS835 | 2457T/ΔospC1F (ospF::cat); CmR | This study |
| BS836 | 2457T/ΔospC1F; unmarked | This study |
| BS838 | 2457T/ΔospBF (ospB::cat); CmR | This study |
| BS873 | BS771 transformed with pBAD24:: ospFMutRb | This study |
| Escherichia coli | ||
| DH5α | endA1 hsdR17 supE44 thi-1 recA1 gyrA96 relA1 (ΔlacIZYA-argF)U169 deoR (Φ80 dlacΔ [lacZ] M15) | Hanahan 1983 |
| BL21 pLysS | F- ompT hsdSB (rB- mB-) dcm gal λ(DE3) pLysS(CmR) tonA | Novagen Inc. |
| DB3.1 | F- gyrA462 endA1 Δ(sr1-recA) mcrB mrr hsdS20(rB-, mB-) supE44 ara14 galK2 lacY1 proA2 rpsL20(SmR) xyl5 Δleu mtl1 | Invitrogen Inc. |
| Plasmids | ||
| pKD3 | bla cat oriR6K; AmpR CmR | Datsenko and Wanner, 2000 |
| pKM208 | Temperature-sensitive red-, gam-, and lacI expressing plasmid driven by PTac promoter; AmpR | Murphy and Campellone, 2003 |
| pCP20 | Plasmid expressing FRT recombinase; AmpR | Datsenko and Wanner, 2000 |
| pDZ2 | Intermediate plasmid with 2HA-tag downstream of cloning site; CmR | Zurawski et al., 2006 |
| pDZ8 | OspB-2HA fusion. Expression driven by POspB; CmR | This study |
| pDONR221 | Donor plasmid for Gateway™ system; KanR | Invitrogen Inc. |
| pDEST-53 | N-terminal GFP destination vector for Gateway® system | Invitrogen Inc. |
| pDEST-53/CAT | GFP alone control. No insert. AmpR, CmR | Invitrogen Inc. |
| pGFP-FL-OspB | ospB cloned into pDEST-53; AmpR | This study |
| pGFP-N-OspB | ospB bp 1-551 cloned into pDEST-53; AmpR | This study |
| pGFP-C-OspB | ospB bp 551-967 cloned into pDEST-53; AmpR | This study |
| pGEX6P-1 | Protein expression vector/GST fusion; AmpR | GE Healthcare Inc. |
| pGST-OspF | ospF cloned into pGEX6P-1; AmpR | Zurawski et al., 2006 |
| pGST-OspB | ospB cloned into pGEX6P-1; AmpR | This study |
| pGST-N-OspF | ospF (bp 1-249) cloned into pGEX6P-1; AmpR | This study |
| pGST-M-OspF | ospF (bp 241-456) cloned into pGEX6P-1; AmpR | This study |
| pGST-C-OspF | ospF (bp 451-724) cloned into pGEX6P-1; AmpR | This study |
| pGST-OspFMutRb | ospF site-directed mutant (IMCLE to RMGLA) cloned into pGEX6P-1; AmpR | This study |
| pBAD24 | Arabinose-inducible vector; AmpR | Guzman et al., 1995 |
| pBAD24::ospF | ospF cloned into pBAD24; AmpR | Zurawski et al., 2006 |
| pBAD24::ospFMutRb | ospFMutRb cloned into pBAD24; AmpR | This study |