Detection of pathogenic bacteria by using zinc finger protein fused with firefly luciferase |
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Authors: | Koichi Abe Takenori Kumagai Chiaki Takahashi Aki Kezuka Yoshiyuki Murakami Yuko Osawa Hiroaki Motoki Takafumi Matsuo Michio Horiuchi Koji Sode Shizunobu Igimi Kazunori Ikebukuro |
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Affiliation: | Department of Biotechnology and Life Science, Tokyo University of Agriculture & Technology , 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan. |
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Abstract: | We constructed a novel bacterial genome detection system using zinc finger protein (ZF) fused with firefly luciferase (ZF-luciferase). Taking advantage of the direct recognition of double-stranded DNA (dsDNA) by ZF, we previously constructed bacterial genome detection systems that did not require dehybridization processes. To detect polymerase chain reaction (PCR) products rapidly and with a high sensitivity, we constructed two kinds of ZF-luciferase, Sp1-fused luciferase (Sp1-luciferase), and Zif268-fused luciferase (Zif268-luciferase). ZF-luciferase not only maintains luciferase activity but also shows dsDNA-binding ability and specificity. Furthermore, we succeeded in the detection of 10 copies of the genome of Legionella pneumophila and Escherichia coli O157. ZF-luciferase would be a useful tool for highly sensitive detection of pathogenic bacterial genome. |
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