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Dissecting iron uptake and homeostasis in Nitrosomonas europaea.

Methods in enzymology (2010-12-28)
Luis A Sayavedra-Soto, Neeraja Vajrala, Daniel J Arp
ANOTACE

The chemolithoautotroph Nitrosomonas europaea oxidizes about 25 mol of NH(3) for each mole of CO(2) that is converted to biomass using an array of heme and nonheme Fe-containing proteins. Hence mechanisms of efficient iron (Fe) uptake and homeostasis are particularly important for this Betaproteobacterium. Among nitrifiers, N.europaea has been the most studied to date. Characteristics that make N.europaea a suitable model to study Fe uptake and homeostasis are as follows: (a) its sequenced genome, (b) its capability to grow relatively well in 0.2 μM Fe in the absence of heterologous siderophores, and (c) its amenability to mutagenesis. In this chapter, we describe the methodology we use in our laboratory to dissect Fe uptake and homeostasis in the ammonia oxidizer N. europaea.

MATERIÁLY
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Sigma-Aldrich
Iron(III) sulfate hydrate, BioReagent, suitable for plant cell culture
Sigma-Aldrich
Ferrous sulfate chelate solution, 100 ×, suitable for plant cell culture