Detection of Pathogenic Bacteria by Using Zinc Finger Protein Fused with Firefly Luciferase

详细信息    查看全文

• 作者：Koichi Abe ; Takenori Kumagai ; Chiaki Takahashi ; Aki Kezuka ; Yoshiyuki Murakami ; Yuko Osawa ; Hiroaki Motoki ; Takafumi Matsuo ; Michio Horiuchi ; Koji Sode ; Shizunobu Igimi ; Kazunori Ikebukuro
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：September 18, 2012
• 年：2012
• 卷：84
• 期：18
• 页码：8028-8032
• 全文大小：254K
• 年卷期：v.84,no.18(September 18, 2012)
• ISSN：1520-6882

文摘

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.