Cloning, expression, purification and characterization of an iron-dependent regulator protein from Thermobifida fusca.

Abstract

Iron-dependent regulators (IdeRs) control the transcription of a variety of genes associated with iron homeostasis in Gram-positive bacteria. In this study we report the cloning of a putative IdeR gene from the moderate thermophile Thermobifida fusca into the pET-21a(+) expression vector. The expressed protein, Tf-IdeR, was purified using immobilized metal affinity and size-exclusion chromatography, and yielded approximately 12-16 mg of protein per liter of culture. The purified Tf-IdeR protein binds the tox operator sequence in the presence of divalent metal ions. Two Tf-IdeR binding sites were identified in the T. fusca genome upstream of a putative enterobactin exporter and a putative ABC-type multidrug transporter.

DOI: 10.1016/j.pep.2013.09.010

5 Figures and Tables

Cite this paper

@article{Granger2013CloningEP, title={Cloning, expression, purification and characterization of an iron-dependent regulator protein from Thermobifida fusca.}, author={Joseph B Granger and Zeyu Lu and Jordan B Ferguson and Peter J Santa Maria and Walter R. P. Novak}, journal={Protein expression and purification}, year={2013}, volume={92 2}, pages={190-4} }