Simultaneous analysis of bacterioferritin gene expression and intracellular iron status in Pseudomonas putida KT2440 by using a rapid dual luciferase reporter assay.
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 (bfralpha, bfrbeta, 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 bfralpha (PP0482) and bfrbeta (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.
Chen
Shicheng
S
Department of Soil Science, University of Wisconsin, Madison, 53706-1299, USA.
Bleam
William F
WF
Hickey
William J
WJ
D001426:Bacterial Proteins / Q000096:biosynthesis*; D003573:Cytochrome b Group / Q000096:biosynthesis*; D005293:Ferritins / Q000096:biosynthesis*; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D007501:Iron / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D016958:Pseudomonas putida / Q000235:genetics* / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D001426:Bacterial Proteins; D003573:Cytochrome b Group; D005293:Ferritins; C018032:bacterioferritin; D007501:Iron; D008156:Luciferases