Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant genotype or descriptionReference or source
Strains
E. coli
  DH5α end-1 hsdR17 (rK mK +)supE44 thi-1 recA1 gyrA (Nalr) relA1 Δ(lacIZYA-argF)U169 deoR[φ80dlacΔ(lacZ)M15]GIBCO-BRL
  XL1-Blue end-1 hsdR17(rK mK +) supE44 thi-1 λ recA1 gyrA96 (Nalr) relA1lac)[F′ proAB+ lacI q ZΔM15::Tn10(Tcr)]Stratagene
Y. pestisa
  KIM5-3001 (parent)Smr; pCD1 (LCR+), pPCP1(Pla+) pMT1 27
  KIM5-3001.5Smr; pCD1 lcrG (Δ39–53)b, pPCP1(Pla+), pMT1 49
  KIM5-3001.6Smr; pCD1 lcrE(Δ48–197), pPCP1(Pla+), pMT1 42
  KIM5-3001.17 (ΔP1)Smr; pCD1yscP(Δ246–333), pPCP1(Pla+), pMT1This study
  KIM5-3001.18 (ΔP2)Smr; pCD1yscP(Δ57–324), pPCP1(Pla+), pMT1This study
  KIM5-3401Smr, pCD1lcrH::cat yopJ::Mu d1734 [Kmr Lac+] [LcrH, YopB, YopD, YopJ], pPCP1, pMT1 42
  KIM8-3002 (parent Pla)Smr; pCD1 (LCR+), pPCP1(Pla), pMT1 34
  KIM8-3002.1Smr, pCD1 yopB(Δ8–399), pPCP1(Pla), pMT1 13
  KIM8-3002.4 (ΔP2-Pla)Smr; pCD1yscP(Δ57–324), pPCP1(Pla) pMT1This study
Plasmids
 pBluescript II SK+Apr; cloning vectorStratagene
 pBluescript II SK−Apr; cloning vectorStratagene
 pYP-F2Apr; BamHI F fragment cloned from pBGCD1 carrying yscN′OPQRS, with frameshift mutation inyscR, was cloned into pBluescript II SK− with the insert oriented with the lac promoter 14
 pYscOP.2Apr; 2.38-kb Bpu1102I fragment of pYP-F2 carrying yscO and yscP was filled in with Klenow and cloned into XhoI digested/Klenow blunt-ended pBluescript II SK+ with the insert oriented with the lacpromoter 37
 pYscOPApr; same as pYscOP.2 with the insert oriented with the T7 promoter 37
 pYscO.2Apr;SphI-KpnI (KpnI site in vector) digestion of pYscOP.2 followed by blunt ending with T4 polymerase and religation, resulting in elimination of yscP and carryingyscO 37
 pYscPApr; 1.8-kb AvaI fragment of pYP-F1 (14) carrying yscP, filled in with Klenow and cloned into the EcoRV site of pBluescript II SK+ with the insert oriented with the T7 promoterThis study
 pYscP.2Apr; same as pYscP but oriented with thelac promoterThis study
 pYscP1Apr;BclI digest and religation of plasmid pYscP, resulting in deletion of yscP (Δ246–333)This study
 pYscP2Apr; pYscOP digested with AgeI, filled in with Klenow followed by digestion with StyI and blunt ending with mung bean nuclease; religation of plasmid resulted in deletion of yscP (Δ57–324)This study
 pHT-VApr; expression vector carryinglcrV; translationally fused to a leader encoding 23 residues, including 6 histidines 15, 33
 pTRCM.2Apr; expression vector carryingyopM behind the trcpromoter 42
 pGEX-3XApr; GST fusion expression vector with the tac promoterPharmacia
 pGST-YscPApr; BamHI (in vector site)-AgeI digest of pGST-YscO (37) filled in with Klenow followed by religation of plasmid (expresses fusion protein of GST and aa 328–455 of YscP)This study
 pUK4134Apr; suicide vector oriR6K oriT cos rpsL 47
 pUKΔP1 XhoI digest of pYscP1 filled in with Klenow followed by SmaI (vector site) digest;XhoI-SmaI fragment cloned into EcoRV site of pUK4134This study
 pUKΔP2 MluI digest of pYscP2 filled in with Klenow followed by digestion withEcoRV (vector site); MluI-EcoRV band cloned into EcoRV site of pUK4134This study
  • a All Y. pestis strains are Pgm (54).

  • b Numbers in parentheses are the amino acids deleted from the protein product.

  • c AvrII sites introduced by site-directed mutagenesis (see Materials and Methods).