Abstract
Next-generation sequencing (NGS) has been applied to various kinds of omics studies, resulting in many biological and medical discoveries. However, high-throughput protein–protein interactome datasets derived from detection by sequencing are scarce, because protein–protein interaction analysis requires many cell manipulations to examine the interactions. The low reliability of the high-throughput data is also a problem. Here, we describe a cell-free display technology combined with NGS that can improve both the coverage and reliability of interactome datasets. This in vitro method is suitable for exploring the interactome networks of transcription factors.
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Acknowledgments
We thank Dr. Masamichi Ishizaka and Dr. Hiroshi Yanagawa for helpful discussions and Takara Bio Inc. for their technical support in preparing samples for 454 sequencing. This research was partially supported by a Grant in Aid for Scientific Research on Innovative Areas “Integrative Systems Understanding of Cancer for Advanced Diagnosis, Therapy and Prevention (No. 4201)” (Grant No. 23134510) of the Ministry of Education, Culture, Sports, Science, and Technology (MEXT), Japan (to S. F. and E. M. S), and a Female Researcher Science Grant from Shiseido Co., Ltd (to E. M. S.).
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Miyamoto-Sato, E. (2014). Next-Generation Sequencing Coupled with a Cell-Free Display Technology for Reliable Interactome of Translational Factors. In: Miyamoto-Sato, E., Ohashi, H., Sasaki, H., Nishikawa, Ji., Yanagawa, H. (eds) Transcription Factor Regulatory Networks. Methods in Molecular Biology, vol 1164. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0805-9_3
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DOI: https://doi.org/10.1007/978-1-4939-0805-9_3
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-0805-9
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