TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Properties | Source or reference |
|---|---|---|
| E. coli K-12 | ||
| DH5α | F− f80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17 (rK− rM+) phoA supE44 λ− thi-1 gyrA96 relA1 | GIBCO-BRL |
| DH5α λpir | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17 (rK− rM+) phoA supE44 thi-1 gyrA96 relA1 λpir | Laboratory stock |
| Y. pestisa | ||
| KIM6 | pPCP1 (40), pMT1 (40) | R. R. Brubaker |
| KIM10 | pMT1 | Laboratory stock |
| KIM8-3241 | pCD1 (lcrV [LcrVΔ18–215]byopJ::MudI1734 [Kmr Lac+]),c pMT1, pOS50 | 9 |
| KIM8-3002 | Smr; pCD1 (Lcr+), pMT1 | 37 |
| KIM8-3002.1 | Smr; pCD1 (yopB [YopBΔ8-390), pMT1 | This study |
| KIM8-3002.2 | Smr; pCD1 (yopD [YopDΔ1–305]), pMT1 | 72 |
| KIM8-3233 | Smr; pCD1 (yopM::lacZYA), pMT1 | 58 |
| KIM8-3233.1 | Smr; pCD1 (yopM::lacZYA lcrE [LcrEΔ48–197]), pMT1 | This study |
| KIM5-3001.2.1 | Smr; pCD1 (yscV [YscVΔ192–343], yopD [YopDΔ1–305]), pPCP1, pMT1 | 72 |
| Plasmids and constructs | ||
| pLD55 | Apr Tcr; suicide vector for allelic exchange | 28 |
| pΔyopB | PCR-amplified DNA from pAW161 containing ΔyopB (deleting nucleotides 23–1169) in pLD55 | This study |
| pUK4134.6 | PstI deletion of lcrE cloned into pUK4134 | 48 |
| pHT-V | PCR-amplified lcrV cloned in pPRoEX-1 in-frame with a 23-residue leader sequence encoding His6 (lcrV) | 9 |
| pHT-VN68 | Subcloned lcrV in pPRoEX-1 with a 29-residue, His6-containing leader sequence fused at the EcoRV site in lcrV (′lcrV [LcrVΔ1–67]) | 9 |
| pTrcV | BbvI/NcoI fragment of pES6–1 cloned into SmaI-cut pTrc99A (lcrV) | 9 |
| TrcM.2 | Subcloned yopM in pTrc99A (yopM) | 48 |
| pAW161 | PCR-amplified lcrH and yopD cloned into pBAD33 (14) expression vector (lcrH, ′yopB [YopBΔ8–390], yopD) | 72 |
| pOS50 | Tcr; competitive plasmid for pPCP1 | J. Goguen |
a
All strains are Pgm− (69).
b
Numbers in brackets list residues deleted from gene product.
c
Descriptions in parentheses give relevant genes present on construct.