Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant propertiesReference or source
E. coli
 BMH 71-18mutS thi supE Δ(lac-proAB) [mutS::Tn10] [F′ proAB laqI q ZΔM15]Promega
 DH5α end-1 hsdR17 (rK mK +) supE44 thi-1 recA1 gyrA(Nalr) relA1Δ(lacIZYA-argF)U169 deoR[φ80dlacΔ(lacZ)M15]GIBCO-BRL
Y. pestisa
 KIM5-3001 (wt)Smr; pCD1 (LCR+), pPCP1(Pla+) pMT1 38
 KIM5-3001.16 (ΔyscO)Smr; pCD1yscO(Δ6–151),bpPCP1(Pla+), pMT1This study
 KIM6-3001Smr; pCD1(LCR), pPCP1(Pla+) 38
 KIM8-3002 (wt Pla)Smr; pCD1, pPCP1(Pla), pMT1 75
 KIM8-3002.3 (ΔyscOPla)Smr; pCD1yscO(Δ6–151),bpPCP1(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 lac promoter 20
 pYscOP.2Apr; 2.38-kb Bpu 1102I 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 thelac promoterThis study
 pYscOPApr; same as pYscOP.2 with insert oriented with the T7 promoterThis study
 pYscO.2Apr; SphI-KpnI (KpnI site in vector) digestion of pYscOP.2 followed by blunt ending with T4 polymerase and religation resulting in the elimination of yscP and carrying yscO This study
 pYscOApr; ClaI (one site in vector) digest of pYscOP followed by religation of plasmid, carriesyscO This study
 pΔyscO Apr;AvrIIc-digested pyscOP.2 was filled in with Klenow and religated, resulting in a deletion ofyscO (Δ6–151)b This study
 pYscN′ΔOPApr; 400-bp SacII fragment of pYP-F2 ligated with ∼5.0-kb SacII fragment of pΔyscO carrying yscN′, yscO(Δ6–151),b and yscP This study
 pYscPApr; 1.8-kb AvaI fragment of pYP-F1 (20) carrying yscP, filled in with Klenow and cloned into EcoRV site of pBluescript II SK+ with insert oriented with the T7 promoterThis study
 pHTVApr; expression vector carryinglcrV; translationally fused to a leader encoding 19 residues, including 6 histidines 21, 45
 pTRCM.2Apr; expression vector carryingyopM behind the trcpromoter 56
 pGEX-3XApr; GST fusion expression vector using the tac promoterPharmacia
 pGST-YscOApr; 1.9-kbMluI-EcoRI (vector site) fragment of pyscO filled in with Klenow following MluI digestion and directionally cloned intoSmaI-EcoRI-digested pGEX-3X (expresses fusion protein of GST and aa 13–154 of YscO)This study
 pUK4134Apr; suicide vector oriR6KoriT cos rpsL 65
 pUKΔyscO Apr; ∼2.3-kbXbaI (in vector site)-AgeI fragment of pyscN′ΔOP filled in with Klenow and cloned into theEcoRV site of pUK4134, carries yscO(Δ16–151)b This study
 pCVD442Apr Sucs; suicide vector 16
 pCVD442ΔyscO Apr Sucs; same insert as in pUKΔyscO cloned into the SmaI site of pCVD442This study
  • a All Y. pestis strains are Pgm (76).

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

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