Table 1.

Strains, plasmids, and primers used

Strain, plasmid, or primerDescriptionReference or sourcea
Strains
Escherichia coli strains
  W3110K-12 FIN(rrnD-rrnE)Rosen
  AW3110W3110 Δars::cam 4
  MC1061K-12 F araD139? galU galK hsdR rpsL Persson
  BH5262K-12araD139? galU galK hsdR rpsL argH1 trxA7004 ilvC::Tn10 gshA 14
  BH2012K-12 araD139? galU galK hsdR rpsL metA46 argH1 trxA7004 ilvC::Tn5 14
Thiobacillus ferrooxidans strains
  ATCC 33020Wild typeATCC
  ATCC 23270Wild typeATCC
  ATCC 19859Wild typeATCC
  ATCC 13598Wild typeATCC
  Thiobacillus caldusMNGWild typeThis laboratory
  Thiobacillus thiooxidans ATCC 19377Wild typeATCC
  Leptospirillum ferrooxidans DSM 2705Wild typeDSM
Plasmids
 pBluescript(SK)Ampr lacZ′, ColE1 replicon, cloning vectorStratagene
 pUCBM21Ampr lacZ′, ColE1 replicon, cloning vectorStratagene
 pACYC184TcrCmr, p15A replicon, cloning vector 6
 pTfars1aApr (from T. ferrooxidansplasmid library, 7-kb Sau3A1 fragment cloned into pEcoR251 digested with BglII)Library of R. S. Ramesar
 pTfars1bApr (6.7-kbHindIII-BglII fragment of pTfars1a cloned into pBluescript)This study
 pTfarsCRBHApr(5.3-kb HindIII-StuI fragment of pTfars1b cloned into pBluescript cut with EcoRV andHindIII)This study
 pTfarsCRBApr (EcoRV-XbaI fragment of pTfars1b, which contained the 5′ end of arsB and all of arsH, was replaced with theEcoRV-XbaI fragment of pBB08, which contained only the 5′ end of arsH)This study
 pTfarsBHApr (KpnI deletion of pTfarsCRBH)This study
 pTfarsBApr(PstI deletion of pTfarsBH)This study
 pTfarsHApr (PCR product obtained with primers ARSHF and ARSHR, cloned into pBluescript)This study
 pTfarsCApr (PCR product obtained with primers ARSCF and ARSR, cloned into pBluescript)This study
 pTfarsRApr (0.55-kb SphI fragment of pTfarsCRBH, cloned into pUCBM21)This study
 pBB08Apr (0.8-kb EcoRV-PstI fragment of pTfars1b, cloned into pBluescript)This study
 pTfarsCRBH-CmCmr (6.7-kbHindIII-BglII fragment of pTfars1a, cloned into the tet gene of pACYC184)This study
 pTrx6Apr (contains T. ferrooxidansATCC 33020 trxA gene, cloned into pBluescript) 19
Primers
 BBARSB(BamHI) 5′-GCGGATCCAGGGTGACGAGAAATATGGC-3′This study
 BBARSC(BamHI) 5′-GCGGATCCGGGGTTTTCATCACTGG-3′This study
 ARSHF(EcoRI) 5′-GCGAATTCTGGTGGCTGCCGCTGGCTTG-3′This study
 ARSHR(HindIII) 5′-GAAAGCTTGCGTACCCCCAACCTCATGCC3′This study
 BBARSCF(HindIII) 5′-GCAAGCTTCGGTGAAACCCGCTCTCCCT-3′This study
 BBARSCR(EcoRI) 5′-GCGAATTCTGTGCGCGCCTTGATGGGTGGC-3′This study
  • a Rosen, B. Rosen, Wayne State University, Detroit, Mich.; Persson, Britt Persson, University of Umea, Umea, Sweden; ATCC, American Type Culture Collection, Rockville, Md.; DSM, Deutsche Sammlung von Mikroorganismen, Braunschweig, Germany; Stratagene, Stratagene, La Jolla, Calif.