Table 1.

Strains and plasmids used in this study

Strain or plasmidCharacteristicsSource or reference
E. coliDH5α supE44 ΔlacU169 (Δ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 Laboratory collection
M. tuberculosis
  H37RvVirulent laboratory strain (ATCC 27294)Laboratory collection
  SJA14Single-crossover (SCO) recombinant with pSA1;relA ΔrelA::hyg lacZHygr This work
  SJA16Double-crossover (DCO) recombinant with pSA1 containing integrated copy of pSA1; ΔrelA::hyg lacZHygr This work
  SJA32SCO recombinant with pSA3; relA+ sacB Hygr This work
  SJA33DCO recombinant obtained with pSA3; ΔrelA::hyg Hygr This work
  CDC1551Virulent M. tuberculosis clinical isolate (CSU93) 68
 pGEM3Z(+)f E. coli cloning vector; Apr Promega
 Pac6Cosmid from pYUB328::H37Rv carrying relA This work
 pIJ963 E. coli cloning vector; AprHygr 29
 pSMT3 E. coli-Mycobacterium shuttle vector carrying mycobacterialhsp60 promoter; Hygr 55
 pSMT3lacZ pSMT3 derivative carrying lacZcloned in BamHI site; hsp60-lacZ This work
 p2NIL E. coli cloning vector with uniquePacI site; Kmr 56
 pGOAL13 E. coli cloning vector withhsp60-sacB cassette carried on PacI fragment; Apr 56
 pRelΔSpGEM3Z(+)f derivative carrying ΔrelA allele lacking internalSphI fragmentThis work
 pRelΔSHpRelΔS derivative carrying ΔrelA marked with hyg gene from pIJ963 inserted in BglII site of ΔrelArelA::hyg)This work
 pSA1pRelΔSH carrying hsp60-lacZ This work
 pSA3p2NIL derivative carrying ΔrelA::hyg allele from pRelΔSH andPacI cassette from pGOAL13; hsp60-sacB; Kmr Hygr This work
 pMV306K E. coli-Mycobacterium shuttle vector; integrates into the L5attB site in Mycobacteria; Kmr 41
 pMV306K-rel pMV306K derivative carryingrelMtb This work
 pMV306K-hsp60-luc pMV306K carrying theNotI-SalI luc fragment from pGL3 (Promega) and the KpnI-HindIIIgroEL promoter from pMV261 16