TABLE 1.
Bacteria and plasmids used in this study
| Bacterial strain or plasmid | Description | Reference(s) | Source |
|---|---|---|---|
| S. putrefaciens | |||
| MR-1 | Manganese-reducing strain from Lake Oneida, N.Y., sediments | 38 | Previous study |
| MR-1A | Spontaneous Rfr mutant of MR-1 | 32 | Previous study |
| MR-1B | Spontaneous Rfr mutant of MR-1 | This work | |
| CMTn-1 | Transposon mutant of MR-1A that is unable to use Fe(III), nitrate, and fumarate as terminal electron acceptors; cymA::TnphoA | 32, 34 | Previous study |
| MR1-CYMA | cymA mutant derived from MR-1; cymA::Kmr | This work | |
| E. coli | |||
| JM109 | recA1 F′ traD36 lacIq Δ(lacZ)M15 proA+B+/e14−(McrA−) Δ(lac-proAB) thi gyrA96 (Nalr) endA1 hsdR17 (rK− mK+) relA1 supE44 | 55 | Promega |
| S17-1 | C600::RP-4 2-Tc::Mu-Km::Tn7 hsdR hsdM+ recA thi pro; donor strain to mate pVK100-derived plasmids into MR-1 | 50 | M. L. P. Collins |
| S17-1λ pir | λ(pir) hsdR pro thi; chromosomally integrated RP4-2 Tc::Mu Km::Tn7; donor strain to mate pEP185.2-derived plasmids into MR-1 | 50 | V. L. Miller |
| TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (Smr) endA1 nupG | Invitrogen | |
| Plasmids | |||
| pVK100 | 23-kb broad-host-range cosmid; Tcr Kmr Tra+ | 19 | ATCC 37156 |
| pGEM-7Zf(+) | 3.00-kb cloning/sequencing vector; Apr | Promega | |
| pCR2.1-TOPO | 3.9-kb vector for cloning PCR products | Invitrogen | |
| pCMTN1-VK | pVK100 with the MR-1 cymA open reading frame plus associated 5′ and 3′ regions | 34 | Previous study |
| pUT/mini-Tn5Km | Apr; Tn5-based delivery plasmid; used as source of Kmr | 10 | D. Frank |
| pTOPO-cymA | pCR2.1-TOPO with the MR-1 cymA open reading frame plus associated 5′ and 3′ regions; 4.9 kb | This work | |
| pTOPO/cymA:Km | pTOPO-cymA with the Kmr gene replacing bases 932–1214 of cymA; 6.7 kb | This work | |
| pEP185.2 | 4.28-kb mobilizable suicide vector derived from pEP184; oriR6K mobRP4 Cmr | 18 | V. L. Miller |
| pDSEPcymAII | Kmr gene-interrupted cymA gene cloned into the SmaI site of pEP185.2; Cmr Kmr; 7.1 kb; used for gene replacement to generate strain MR1-CYMA | This work |