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Signal Enhancement by a Multi-layered Substrate for Mutagen Detection Using an SOS Response-induced Green Fluorescent Protein in Genetically Modified Escherichia coli

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Abstract

In this paper, we describe a method to enhance the fluorescence signal of mutagen detection using SOS response-induced green fluorescence protein (GFP) in genetically modified Escherichia coli using a multi-layered substrate. To generate E. coli that express SOS response-induced GFP, we constructed a plasmid carrying the RecA promoter located upstream of the GFP gene and used it to transform E. coli BL21. The transformed strain was incubated with mitomycin C (MMC), a typical mutagen, and then immobilized on a multi-layered substrate with Ag and a thin Al2O3 layer on a glass slide. Since the multi-layered substrate technique is an optical technique with potential to enhance the fluorescence of fluorophore placed on top of the substrate, the multi-layered substrate was expected to improve the fluorescence signal of mutagen detection. We obtained an average 14-fold fluorescence enhancement of MMC-induced GFP in the concentration range 1 to 1000 ng/ml. In addition, the lower detection limit of MMC was improved using this technique, and was estimated to be 1 ng/ml because of an enlargement of the difference between the blank and the signal of 1 ng/ml of MMC.

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Authors and Affiliations

  1. School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachiouji, Tokyo, 192-0982, Japan

    Hiroki Etoh, Mitsuru Yasuda & Takuo Akimoto

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  1. Hiroki Etoh
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  2. Mitsuru Yasuda
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  3. Takuo Akimoto
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Correspondence to Takuo Akimoto.

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Etoh, H., Yasuda, M. & Akimoto, T. Signal Enhancement by a Multi-layered Substrate for Mutagen Detection Using an SOS Response-induced Green Fluorescent Protein in Genetically Modified Escherichia coli. ANAL. SCI. 27, 1179–1183 (2011). https://doi.org/10.2116/analsci.27.1179

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  • DOI: https://doi.org/10.2116/analsci.27.1179

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