TABLE 1.
Strains, plasmids, and primers used
| Strain, plasmid, or primer | Description | Reference or sourcea |
|---|---|---|
| Strains | ||
| Escherichia coli strains | ||
| W3110 | K-12 F− IN(rrnD-rrnE) | Rosen |
| AW3110 | W3110 Δars::cam | 4 |
| MC1061 | K-12 F−araD139? galU galK hsdR rpsL | Persson |
| BH5262 | K-12 araD139? galU galK hsdR rpsL argH1 trxA7004 ilvC::Tn10 gshA | 14 |
| BH2012 | K-12 araD139? galU galK hsdR rpsL metA46 argH1 trxA7004 ilvC::Tn5 | 14 |
| Thiobacillus ferrooxidans strains | ||
| ATCC 33020 | Wild type | ATCC |
| ATCC 23270 | Wild type | ATCC |
| ATCC 19859 | Wild type | ATCC |
| ATCC 13598 | Wild type | ATCC |
| Thiobacillus caldus MNG | Wild type | This laboratory |
| Thiobacillus thiooxidans ATCC 19377 | Wild type | ATCC |
| Leptospirillum ferrooxidans DSM 2705 | Wild type | DSM |
| Plasmids | ||
| pBluescript(SK) | AmprlacZ′, ColE1 replicon, cloning vector | Stratagene |
| pUCBM21 | AmprlacZ′, ColE1 replicon, cloning vector | Stratagene |
| pACYC184 | Tcr Cmr, p15A replicon, cloning vector | 6 |
| pTfars1a | Apr (from T. ferrooxidans plasmid library, 7-kb Sau3A1 fragment cloned into pEcoR251 digested with BglII) | Library of R. S. Ramesar |
| pTfars1b | Apr (6.7-kb HindIII-BglII fragment of pTfars1a cloned into pBluescript) | This study |
| pTfarsCRBH | Apr (5.3-kb HindIII-StuI fragment of pTfars1b cloned into pBluescript cut with EcoRV and HindIII) | This study |
| pTfarsCRB | Apr (EcoRV-XbaI fragment of pTfars1b, which contained the 5′ end of arsB and all of arsH, was replaced with the EcoRV-XbaI fragment of pBB08, which contained only the 5′ end of arsH) | This study |
| pTfarsBH | Apr (KpnI deletion of pTfarsCRBH) | This study |
| pTfarsB | Apr (PstI deletion of pTfarsBH) | This study |
| pTfarsH | Apr (PCR product obtained with primers ARSHF and ARSHR, cloned into pBluescript) | This study |
| pTfarsC | Apr (PCR product obtained with primers ARSCF and ARSR, cloned into pBluescript) | This study |
| pTfarsR | Apr (0.55-kb SphI fragment of pTfarsCRBH, cloned into pUCBM21) | This study |
| pBB08 | Apr (0.8-kb EcoRV-PstI fragment of pTfars1b, cloned into pBluescript) | This study |
| pTfarsCRBH-Cm | Cmr (6.7-kb HindIII-BglII fragment of pTfars1a, cloned into the tet gene of pACYC184) | This study |
| pTrx6 | Apr (contains T. ferrooxidans ATCC 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.