Abstract
The newly developed Transdirect in vitro translation system for mRNA templates utilizes an extract from cultured Spodoptera frugiperda 21 (Sf21) insect cells. An expression vector, pTD1, which includes a 5′-untranslated region (UTR) sequence from a baculovirus polyhedrin gene as a translational enhancer, designed to obtain maximum performance from the insect cell-free protein synthesis system. The combination of insect cell extract and the expression vector results in protein productivity of about 50 μg/mL of the translation reaction mixture. This is the highest protein productivity yet recorded among commercialized cell-free protein synthesis systems based on animal extracts.
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References
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Acknowledgements
We wish to thank Dr. Osamu Ohara and Dr. Takahiro Nagase, Kazusa DNA Research Institute, for valuable discussions. We would also like to thank Dr. Sumiharu Nagaoka, Kyoto Institute of Technology; Dr. Toshihiko Utsumi, University of Yamaguchi; and Dr. Susumu Tsunasawa, Shimadzu Corporation, for helpful discussions.
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Ezure, T., Suzuki, T., Ando, E. (2014). A Cell-Free Protein Synthesis System from Insect Cells. In: Alexandrov, K., Johnston, W. (eds) Cell-Free Protein Synthesis. Methods in Molecular Biology, vol 1118. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-782-2_20
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DOI: https://doi.org/10.1007/978-1-62703-782-2_20
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-62703-782-2
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