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J. Microbiol. Biotechnol. 2006; 16(3): 386-390

Published online March 28, 2006

Copyright © The Korean Society for Microbiology and Biotechnology.

Construction of a Reporter Strain Pseudomonas putida for the Detection of Oxidative Stress Caused by Environmental Pollutants

Lee, Yunho , Eunyoung Ahn 1, Sungsu Park 1, Eugene L. Madsen 2, Che Ok Jeon 3 and Woojun Park 4*

Division of Environmental Science and Ecological Engineering, Korea University, Seoul 136-701, Korea, 1Division of Nano Sciences, Ewha Women’s University, Seoul 120-750, Korea, 2Department of Microbiology, Cornell University, Ithaca, NY 14853, U.S.A., 3Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea, 4Division of Environmental Science and Ecological Engineering, Korea University, Seoul 136-701, Korea Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea

Abstract

A green fluorescent protein-based Pseudomonas putida reporter was successfully constructed and shown to be capable of detecting oxidative stress. In this whole-cell reporter, the promoter of the paraquat-inducible ferredoxin-$NADP^+$ reductase (fpr) was fused to a promoterless gfp gene on a broad-host-range promoter probe vector. Pseudomonas putida KT2440 harboring this reporter plasmid exhibited an increased level of gfp expression in the presence of redox-cycling agents (paraquat and menadione), hydrogen peroxide, and potential environmental pollutant chemicals such as toluene, paint thinner, gasoline, and diesel. Induction of fpr in the presence of these chemicals was confirmed using Northern blot analysis.

Keywords: Paraquat, menadione, paint thinner, hydrogen peroxide, green fluorescence protein