Signal enhancement by a multi-layered substrate for mutagen detection using an SOS response-induced green fluorescent protein in genetically modified Escherichia coli

Hiroki Etoh, Mitsuru Yasuda, Takuo Akimoto

研究成果: Article査読

5 被引用数 (Scopus)

抄録

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 Al 2 O 3 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. 2011

本文言語English
ページ(範囲)1179-1183
ページ数5
ジャーナルanalytical sciences
27
12
DOI
出版ステータスPublished - 2011
外部発表はい

ASJC Scopus subject areas

  • Analytical Chemistry

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