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Nucleic Acids Symposium Series 2006 50(1):277-278; doi:10.1093/nass/nrl138
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© 2006 Oxford University Press

Efficient incorporation of a nonnatural amino acid into a protein in an insect cell-free translation system

Yasunori Tokuda, Masumi Taki and Masahiko Sisido

Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan

Recently, we have succeeded in incorporating various nonnatural amino acids into proteins by using four-base codon-anticodon pairs in Spodoptera frugiperda 21 (Sf21) insect cell-free protein synthesizing system. Here, the reaction was conducted under various conditions in order to optimize the incorporation efficiency. The optimal concentration of aminoacyl-tRNA, reaction temperature, and reaction time were 2 nM, 25 °C, and 1.5 hr, respectively.


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