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Nucleic Acids Research Advance Access originally published online on May 23, 2008
Nucleic Acids Research 2008 36(11):e68; doi:10.1093/nar/gkn274
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Nucleic Acids Research, 2008, Vol. 36, No. 11 e68
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

The simple and rapid detection of specific PCR products from bacterial genomes using Zn finger proteins

Yuko Osawa1, Kazunori Ikebukuro1,*, Hiroaki Motoki2, Takafumi Matsuo2, Michio Horiuchi2 and Koji Sode1

1Department of Biotechnology and Life Science, Tokyo University of Agriculture & Technology, 2-24-16 Naka-cho, Koganei, 184-8588 Tokyo and 2SYSTEM INSTRUMENTS Co., Ltd, 776-2 Komiya-cho, Hachioji, 192-0031 Tokyo, Japan

*To whom correspondence should be addressed. Tel: +81 42 388 7030; Fax: +81 42 388 7030; Email: ikebu{at}cc.tuat.ac.jp

Received October 3, 2007. Revised April 16, 2008. Accepted April 25, 2008.

A novel method of rapid and specific detection of polymerase chain reaction (PCR) products from bacterial genomes using Zn finger proteins was developed. Zn finger proteins are DNA-binding proteins that can sequence specifically recognize PCR products. Since Zn finger proteins can directly detect PCR products without undergoing dehybridization, unlike probe DNA, and can double check the specific PCR amplification and sequence specificity of the PCR products, this novel method would be quick and highly accurate. In this study, we tried to detect Legionella pneumophila using Sp1. It was found that a 49 bp L. pneumophila-specific region containing the Sp1 recognition site is located on the flhA gene of the L. pneumophila genome. We succeeded in specifically detecting PCR products amplified from L. pneumophila in the presence of other bacterial genomes by ELISA, and demonstrated that Sp1 enables the discrimination of L. pneumophila-specific PCR products from others. By fluorescence depolarization measurement, these specific PCR products could be detected within 1 min. These results indicate that the rapid and simple detection of PCR products specific to L. pneumophila using a Zn finger protein was achieved. This methodology can be applied to the detection of other bacteria using various Zn finger proteins that have already been reported.


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