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Journal of Biochemistry 2005 137(2):121-124; doi:10.1093/jb/mvi023
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© 2005 The Japanese Biochemical Society

BIOTECHNOLOGY

Application of Quantitative Real-Time PCR for Monitoring the Process of Enrichment of Clones on In Vitro Protein Selection

Kenichi Horisawa, Nobuhide Doi, Hideaki Takashima and Hiroshi Yanagawa1

Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522

1 To whom correspondence should be addressed. Tel: +81 45 566 1775, Fax: +81 45 566 1440, E-mail: hyana{at}bio.keio.ac.jp

In vitro selection of proteins from cDNA libraries using display technologies, such as the in vitro virus method, is a powerful means for the discovery of novel protein interactions. After iterative screening, selected proteins are usually identified and evaluated by cloning and sequencing analysis. Previously we applied real-time PCR for evaluation of the sequences obtained on in vitro virus screening. Here, we have presented additional data regarding monitoring of the process of enrichment of selected clones in each round of selection and elimination of false positives by real-time PCR, and have also discussed the utility of the novel method. This approach should also be applicable to other display technologies.


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