Abstract
The Transdirect insect cell is a newly developed in vitro translation system for mRNA templates, which 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, was also developed to obtain maximum performance from the insect cell-free protein synthesis system. This combination of insect cell extract and expression vector results in protein productivity of about 50 μg/mL of the translation reaction mixture. This is the highest protein productivity yet noted among commercialized cell-free protein synthesis systems based on animal extracts.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ezure, T., Suzuki, T., Higashide, S., Shintani, E., Endo, K., Kobayashi, S., Shikata, M., Ito, M., Tanimizu, K., and Nishimura, O. (2006) Cell-free protein synthesis system prepared from insect cells by freeze-thawing. Biotechnol. Prog. 22, 1570–1577.
Suzuki, T., Ito, M., Ezure, T., Shikata, M., Ando, E., Utsumi, T., Tsunasawa, S., and Nishimura, O. (2006) N-Terminal protein modifications in an insect cell-free protein synthesis system and their identification by mass spectrometry. Proteomics 6, 4486–4495.
Suzuki, T., Ito, M., Ezure, T., Shikata, M., Ando, E., Utsumi, T., Tsunasawa, S., and Nishimura, O. (2007) Protein prenylation in an insect cell-free protein synthesis system and identification of products by mass spectrometry. Proteomics 7, 1942–1950.
Suzuki, T., Ito, M., Ezure, T., Kobayashi, S., Shikata, M., Tanimizu, K., and Nishimura, O. (2006) Performance of expression vector, pTD1, in insect cell-free translation system. J. Biosci. Bioeng. 102, 69–71.
Acknowledgments
We thank Dr. Osamu Ohara and Dr. Takahiro Nagase, Kazusa DNA Research Institute, for valuable discussions. We also thank Dr. Sumiharu Nagaoka, Kyoto Institute of Technology, Dr. Toshihiko Utsumi, University of Yamaguchi, and Dr. Susumu Tsunasawa, Shimadzu Corporation, for helpful discussions.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Ezure, T., Suzuki, T., Shikata, M., Ito, M., Ando, E. (2010). A Cell-Free Protein Synthesis System from Insect Cells. 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_4
Download citation
DOI: https://doi.org/10.1007/978-1-60327-331-2_4
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60327-330-5
Online ISBN: 978-1-60327-331-2
eBook Packages: Springer Protocols