Abstract
Cell-free protein synthesis is a suitable protein expression method for the high-throughput use because a PCR-amplified linear DNA fragment is utilized as a template for protein synthesis without any cloning procedures. We have developed a two-step PCR method for high-throughput and robust production of linear templates ready for cell-free protein synthesis. A high-throughput protein expression method has been established by combining the batch-mode cell-free protein synthesis with the two-step PCR, which is performed on multiwell plates, and is thus adapted for robotics. In this chapter, our two-step PCR method and the batch-mode cell-free protein synthesis are described.
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Acknowledgments
The author would like to thank Takashi Yabuki, Satoru Watanabe, Yoko Motoda, and Kazuharu Hanada for developing our two-step PCR method.
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Kigawa, T. (2010). Analysis of Protein Functions Through a Bacterial Cell-Free Protein Expression System. In: Endo, Y., Takai, K., Ueda, T. (eds) Cell-Free Protein Production. Methods in Molecular Biology, vol 607. Humana Press. https://doi.org/10.1007/978-1-60327-331-2_6
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DOI: https://doi.org/10.1007/978-1-60327-331-2_6
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