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Applied and Environmental Microbiology, February 2009, p. 866-868, Vol. 75, No. 3
0099-2240/09/$08.00+0     doi:10.1128/AEM.01823-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Simultaneous Analysis of Bacterioferritin Gene Expression and Intracellular Iron Status in Pseudomonas putida KT2440 by Using a Rapid Dual Luciferase Reporter Assay{triangledown} ,{dagger}

Shicheng Chen, William F. Bleam, and William J. Hickey*

Department of Soil Science, University of Wisconsin—Madison, Madison, Wisconsin

Received 6 August 2008/ Accepted 22 November 2008

A dual luciferase reporter (DLR) system utilizing firefly and Renilla luciferases was developed and tested in a model rhizobacterium, Pseudomonas putida KT2440. The DLR was applied to simultaneously analyze expression of three putative bacterioferritin genes (bfr{alpha}, bfrβ, and bfr) and assess the cellular iron status of strain KT2440 by monitoring expression of the Fur-regulated fepA-fes promoter. The DLR proved to be reproducible and sensitive. Expression of bfr{alpha} (PP0482) and bfrβ (PP1082) was consistent with expectations for bacterioferritin and varied directly with the iron level. However, expression of bfr (PP4856) was inversely related to the iron concentration and it was thus more likely to encode a Dps-like protein rather than a bacterioferritin.

* Corresponding author. Mailing address: Department of Soil Science, University of Wisconsin, 1525 Observatory Dr., Madison, WI 53706-1299. Phone: (608) 262-9018. Fax: (608) 265-2595. E-mail: wjhickey{at}wisc.edu

{triangledown} Published ahead of print on 1 December 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, February 2009, p. 866-868, Vol. 75, No. 3
0099-2240/09/$08.00+0     doi:10.1128/AEM.01823-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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