Table 1.

Bacterial strains and plasmids used in this study

Bacterial strain or plasmidDescriptionaReferenceSource
Strains
S. putrefaciens
  MR-1Manganese-reducing strain from sediments of Lake Oneida, N.Y.32Previous study
  ETRA-153 etrA mutant derived from MR-1;etrA::Kmr This work
  METR-1Original study: MR-1 butetrA::pACYC184 Tcr; this study: MR-1 containing freely replicating pACYC184 plus partial etrAsequence from MR-141K. H. Nealson
E. coli
  JM109 recA1 F′traD36 lacI qΔ(lacZ)M15proA + B +/e14(McrA) Δ(lac-proAB) thi gyrA96 (Nalr)endA1 hsdR17(rK mK +) relA1 supE44 49Promega
  S17-1λpir λ(pir) hsdR pro thi; chromosomally integrated RP4-2 Tc::Mu Km::Tn7; donor strain to mate pEP185.2-derived plasmids into MR-145V. L. Miller
  S17-1C600::RP-4 2-Tc::Mu-Km::Tn7 hsdR hsdM + recA thi pro; donor strain to mate pVK100-derived plasmids into MR-145M. L. P. Collins
  TOP10F mcrAΔ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL(Smr) endA1 nupG; used as host for plasmids derived from pCR2.1-TOPOInvitrogen
Plasmids
 pVK10023-kb broad-host-range cosmid; Tcr Kmr Tra+ 11ATCC 37156
 pVKetrApVK100 with the MR-1 etrA open reading frame plus associated 5′ and 3′ regionsThis work
 pCR2.1-TOPO3.9-kb vector for cloning PCR products; Kmr Apr Invitrogen
 pUT/mini-Tn5Km Apr; Tn5-based delivery plasmid; used as source of Kmr gene3D. Frank
 pTOPO/etrApCR2.1-TOPO with the MR-1 etrA open reading frame plus associated 5′ and 3′ regions; 5.4 kbThis work
 pTOPO/etrA:KmpTOPO-etrA with the Kmr gene replacing bases 255–501 of etrA; 7.3 kbThis work
 pEP185.24.28-kb mobilizable suicide vector derived from pEP184; oriR6K mobRP4 Cmr 10V. L. Miller
 pDSEPetrAKmr gene-interruptedetrA gene cloned into the SmaI site of pEP185.2; Cmr Kmr; 7.7 kb; used for gene replacement to generate strain ETRA-153This work
 pACYC1844.24-kb vector; Cmr Tcr 1New England Biolabs
 pCALc5.8-kb vector used for cloning and sequencing; Apr 47Stratagene
  • a Nalr, Smr, Apr, and Cmr, resistance to nalidixic acid, streptomycin, ampicillin, and chloramphenicol, respectively.