Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant phenotype or genotypeReference or origin
E. coli strains
 DH5α supE44ΔlacU169 (φ80lacZΔM15) hsdR17 recA1 gyrA96 thi-1 relA1 Gibco-BRL, Gaithersburg, Md.
 S17-1Smr Spr hdsRRP4-2 kan::Tn7 tet::Mu; integrated in the chromosome 45
 C600 supE44 hsdR thi-1 leuB6 lacY1 tonA21 49
 DA258CmrKmr Tcr (C600)grpE +::Ω-Cmthr::Tn10 2
 DA259Cmr Kmr Tcr (C600) ΔgrpE::Ω-Cmthr::Tn10 2
 BL21/pLysSCmr F ompT(rB mB ) T7lys 46
B. japonicum strainsa
 110spc4Spr (wild type) 37
 5549SprKmr hrcA::kan;hrcA and kan oriented in opposite directionsThis study
Plasmids
 pBluescript II KS+Apr Stratagene
 pBluescript II SK+Apr Stratagene
 pBSL15AprKmr 1
 pUC18Apr 34
 pSUP202Apr Cmr Tcr oriT from RP4 45
 pSUP202pol4Tcr (pSUP202); part of polylinker from pBluescript II KS+ between EcoRI andPstI 10
 pNM481Apr (pUC8) ′lacZ 24
 pNM482Apr (pUC8) ′lacZ 24
 pSUP481Tcr (pSUP202pol4) ′lacZ (5.1-kbEcoRI-StuI fragment from pNM481)H. M. Fischer, unpublished data
 pSUP482Tcr(pSUP202pol4) ′lacZ (5.1-kb EcoRI-StuI fragment from pNM482)H. M. Fischer, unpublished data
 pET28a(+)Kmr Novagene, Madison, Wis.
 pWSK29Apr; low-copy-number vector 48
 pBW401Apr(pWSK29); 1.3-kb SalI-EcoRI fragment containingE. coli grpE 49
 pRJ5099Apr (pRJ9519); 0.9-kbBglII-EcoRI fragment containing the B. japonicum dnaKJ promoter region 30
 pRJ5400Tcr (pSUP202pol4)hspA′-′lacZ 29
 pRJ5523b Apr (pUC18); 3.9-kb EcoRI fragment containing B. japonicum ′hrcA, grpE, and ′dnaK opposite to αlacZ orientationThis study
 pRJ5530b Apr (pBluescript II SK+); 3.0-kb NotI fragment containing B. japonicum hrcA and grpE opposite to αlacZorientationThis study
 pRJ5542Apr (pRJ9519); 0.5-kb NotI-NruI fragment from pRJ5530 containing the B. japonicum hrcA promoter regionThis study
 pRJ5543Apr (pRJ9519); 0.7-kbSacI-FspI fragment from pRJ5530 containing theB. japonicum grpE promoter regionThis study
 pRJ5549Tcr Kmr (pSUP202pol4)hrcA::kan; hrcA andkan oriented in opposite directionsThis study
 pRJ5552Kmr [pET28a(+)]; 1.1-kbNotI-NdeI amplification product containingB. japonicum hrcA gene and plasmid for overexpression of H6-HrcAThis study
 pRJ5554Tcr(pSUP482) grpE′-′lacZ; 348-bpMscI-FspI grpE′ fragment from pRJ5530This study
 pRJ5556Apr (pUC18); 198-bp amplification product containing the promoter region of B. japonicum groESL 4 This study
 pRJ5558Apr (pUC18); 265-bp amplification product containing the promoter region of B. japonicum groESL 5 This study
 pRJ5559Tcr(pSUP481) hrcA′-′lacZ; 937-bp EcoRIhrcA′ fragment from pRJ5530This study
 pRJ7998Tcr (pSUP202pol4)groEL 2′-′lacZ 10
 pRJ8067Tcr (pSUP202pol4)groEL 1′-′-lacZ 3
 pRJ8092Tcr (pSUP202pol4)groEL 5′-′lacZ 3
 pRJ8174Tcr (pSUP202pol4)dnaK′-′lacZ 23
 pRJ8548Tcr (pSUP202pol4)groEL 4′-′lacZ 3
 pRJ9519Apr (pBluescript II SK+) containing theB. japonicum rrn terminator 30
  • a Strains containing chromosomally integrated ′lacZ fusions are listed in Table 3.

  • b The insert of this plasmid is depicted in Fig. 1.