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Cell-Free Protein Synthesis Systems with Extracts from Cultured Human Cells

  • Satoshi Mikami
  • Tominari Kobayashi
  • Hiroaki Imataka
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 607)

Abstract

Cell-free protein synthesis systems have been established with extracts from cultured human cells, HeLa, and hybridoma cells. The former cell line is used to prepare extracts for robust translation, whereas the extract from the latter is primarily employed for expression of glycoproteins. Productivity of both systems can be enhanced by addition of K3L and GADD34, factors that diminish phosphorylation of eIF2α. The coupled transcription/translation system is also available as a convenient tool, particularly for the production of large recombinant proteins.

Key words

Human cells Translation initiation Glycosylation Hybridoma HeLa cells GADD34 K3L Coupled transcription/translation 

References

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    Kobayashi T, Mikami S, Yokoyama S, Imataka H. (2007) An improved cell-free system for picornavirus synthesis. J. Virol. Methods 142, 182–188.CrossRefPubMedGoogle Scholar
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    Mikami S, Kobayashi T, Yokoyama S, Imataka H. (2006) A hybridoma-based in vitro translation system that efficiently synthesizes glycoproteins. J. Biotechnol. 127, 65–78.CrossRefPubMedGoogle Scholar
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    Mikami S, Masutani M, Sonenberg N, Yokoyama S, Imataka H. (2006) An efficient mammalian cell-free translation system supplemented with translation factors. Protein Expr. Purif. 46, 348–357.CrossRefPubMedGoogle Scholar
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    Mikami S, Kobayashi T, Masutani M, Yokoyama S, Imataka H. (2008) A human cell-derived in vitro coupled transcription/translation system optimized for production of recombinant proteins. Protein Expr. Purif. 62, 190–198.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Satoshi Mikami
    • 1
  • Tominari Kobayashi
    • 1
  • Hiroaki Imataka
    • 2
  1. 1.RIKEN Systems and Structural Biology CenterYokohamaJapan
  2. 2.Department of Materials Science and Chemistry, Graduate School of EngineeringUniversity of HyogoHyogoJapan

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