Table 1 Bacterial strains and plasmids used in this study^a

S. oneidensis strains

MR-1

Manganese-reducing strain from Lake Oneida, N.Y. sediments

[1]

OMP35-1

omp35 mutant derived from MR-1; omp35::Km^r

This work

OMCA-1

omcA mutant derived from MR-1; omcA::Km^r

[8]

OMCB-1

omcB mutant derived from MR-1; omcB::Km^r

[8]

ETRA-153

etrA mutant derived from MR-1; etrA::Km^r

[17]

CMA-1

menaquinone-minus mutant derived from MR-1

[6]

E. coli strains

JM109

recA1 F' traD36 lacI^qΔ(lacZ)M15 proA⁺B⁺/ e14^-(McrA^-) Δ(lac-proAB) thi gyrA96 (Nal^r) endA1 hsdR17 (r_K^- m_K⁺) relA1 supE44

[61]

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-1

[62]

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

[62]

TOP10

F^- mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZ ΔM15ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (Sm^r) endA1nupG; used as host for plasmids derived from pCR2.1-TOPO

Invitrogen

Plasmids^b

pBCSK

3.4-kb vector; pUC ori; Cm^r; used for complementation

Stratagene

pBComp218F & R

pBCSK with the MR-1 omp35 open reading frame plus associated 5' and 3' regions; 218 bp upstream sequence in the forward (F) and reverse (R) orientation

This work

pBComp411F & R

pBCSK with the MR-1 omp35 open reading frame plus associated 5' and 3' regions; 411 bp upstream sequence in F and R orientation

This work

pCR2.1-TOPO

3.9-kb vector for cloning PCR products; Km^r Ap^r

Invitrogen

pUT/mini-Tn5Km

Ap^r; Tn5-based delivery plasmid; used as source of Km^rgene

[63]

pTOPO/omp35

pCR2.1-TOPO with the MR-1 omp35 open reading frame plus associated 5' and 3' regions; 5.6 kb

This work

pTOPO/omp35:Km

pTOPO/omp35 with the Km^r gene replacing bases 458–850 of omp35; 7.3 kb

This work

pEP185.2

4.28-kb mobilizable suicide vector derived from pEP184; oriR6K mobRP4 Cm^r

[64]

pDSEPomp35

Km^r-interrupted omp35 gene cloned into the Xho I site of pEP185.2; Cm^r Km^r; 7.7 kb; used for gene replacement to generate strain OMP35-1

This work