Table 1.

Bacteria, plasmid vectors, and pas-containing constructs used in this study

Strain, plasmid, or constructGenotype or descriptionaReference
E. coli strains
 JM105 thi rpsL endA sbcB15 hspR4Δ(lac-proAB) [F′ traD36 proAB lacI qΔM15] 15
 CSH50-Iq rpsL Δ(lac-pro) [F′ traD36 proAB lacI qΔM15] 13
 pMC1403ColE1 replicon, Apr lacZYA* 2
 pACYC184p15a replicon, Cmr Tcr 3
 pKK223-3ColE1 replicon, Apr tac 1
 pOU82R1 replicon, Apr lacZYA 6
Constructs containing pas genes
 pP2HpMC1403 vector, Apr, pas region 1240–1362b This work
 pTac-pasA-pACYCpACYC184 vector, Cmr,pas region 1316–1559This work
 pTac-pasB-pACYCpACYC184 vector, Cmr,pas region 1518–1816This work
 pTac-pasC-pACYCpACYC184 vector, Cmr,pas region 1789–2028This work
 pTac-pasAB-pACYCpACYC184 vector, Cmr,pas region 1316–1816This work
 pTac-pasABC-pACYCpACYC184 vector, Cmr,pas region 1316–2028This work
 pKmM0pTF-FC2 replicon, Kmr, 1–4910c 12
 pKmM1pTF-FC2 replicon, Kmr, 1–4911pasA 12
 pKmM2pTF-FC2 replicon, Kmr, 1–4911pasB 12
 pKmM3pTF-FC2 replicon, Kmr, 1–4911pasC 12
 pKmM1del1d pTF-FC2 replicon, Kmr, 1–4911, Δ1217–1321e 12
 pKmM1del2d pTF-FC2 replicon, Kmr, 1–4911, Δ1232–1943e 12
 pOU-pasABCpOU82 R1 replicon, Apr, pasregion 1158–2027 12
 pOU-tac-pasABCpOU82 R1 replicon, Apr,tac, pas region 1158–2027This work
  • a Apr, ampicillin resistance; Kmr, kanamycin resistance; Cmr, chloramphenicol resistance; Tcr, tetracycline resistance;lacZYA*, lacZYA genes with deletion oflacZ promoter and ATG; tac, trp-lachybrid promoter.

  • b The pas region numbers are the nucleotide positions relative to those of theClaI-PstI fragment of pTV100 (5).

  • c Entire pTF-FC2 replicon from theClaI site to the PstI site of pTV100 including the pas region (5).

  • d Spontaneous deletions of the toxic plasmid pKmM1 (12).

  • e Regions missing from the spontaneous deletions after PasA antidote inactivation (12).