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The Wheat Germ Cell-Free Expression System

Methods for High-Throughput Materialization of Genetic Information

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Chemical Genomics

Abstract

This chapter contains protocols for high-throughput protein production based on the cell-free system prepared from eukaryote wheat embryos.

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References

  1. Blaschke, U. K., Silberstein, J., and Muir, T. W. (2000) Protein engineering by expressed protein ligation. Methods Enzymol. 328, 478–496.

    Article  PubMed  CAS  Google Scholar 

  2. Henrich, B., Lubitz, W., and Plapp, R. (1982) Lysis of Escherichia coli by induction of cloned phi X174 genes. Mol. Gen. Genet. 185, 493–497.

    Article  PubMed  CAS  Google Scholar 

  3. Golf, S. A. and Goldberg, A. L. (1987) An increased content of protease La, the lon gene product, increases protein degradation and blocks growth in Escherichia coli. J. Biol. Chem. 262, 4508–4515.

    Google Scholar 

  4. Chrunyk, B. A., Evans, J., Lillquist, J., Young, P., and Wetzel, R. (1993) Inclusion body formation and protein stability in sequence variants of interleukin-1 beta. J. Biol. Chem. 268, 18,053–18,061.

    PubMed  CAS  Google Scholar 

  5. Kurland, C. G. (1982) Translational accuracy in vitro. Cell 28, 201–202.

    Article  PubMed  CAS  Google Scholar 

  6. Pavlov, M. Y. and Ehrenberg, M. (1996) Mutants of EF-Tu defective in binding aminoacyl-tRNA. Arch. Biochem. Biophys. 328, 9–16.

    Article  PubMed  CAS  Google Scholar 

  7. Roberts, B. E. and Paterson, B. M. (1973) Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proc. Natl. Acad. Sci. USA 70, 2330–2334.

    Article  PubMed  CAS  Google Scholar 

  8. Wool, I. G., Glück, A., and Endo, Y. (1992) Ribotoxin recognition of ribosomal RNA and a proposal for the mechanism of translocation. Trends Biochem. Sci. 17, 266–269.

    Article  PubMed  CAS  Google Scholar 

  9. Barbieri, L., Battelli, M. G., and Stirpe, F. (1993) Ribosome-inactivating proteins from plants. Biochim. Biophys. Acta 1154, 237–282.

    PubMed  CAS  Google Scholar 

  10. Ogasawara, T., Sawasaki, T., Morishita, R., Ozawa, A., Madin, K., and Endo, Y. (1999) A new class of enzyme acting on damaged ribosomes: ribosomal RNA apurinic site specific lyase found in wheat germ. EMBO J. 18, 6522–6531.

    Article  PubMed  CAS  Google Scholar 

  11. Madin, K., Sawasaki, T., Ogasawara, T., and Endo, Y. (2000) A highly efficient and robust cell-free protein synthesis system prepared from wheat embryos: plants apparently contain a suicide system directed at ribosomes. Proc. Natl. Acad. Sci. USA 97, 559–564.

    Article  PubMed  CAS  Google Scholar 

  12. Sawasaki, T., Ogasawara, T., Morishita, R., and Endo, Y. (2002) A cell-free protein synthesis system for high-throughput proteomics. Proc. Natl. Acad. Sci. USA 99, 14,652–14,657.

    Article  PubMed  CAS  Google Scholar 

  13. Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  14. Gurevich, V. V. (1996) Use of bacteriophage RNA polymerase in RNA synthesis. Methods Enzymol. 275, 382–397.

    Article  PubMed  CAS  Google Scholar 

  15. Hanes, J. and Pluckthun, A. (1997) In vitro selection and evolution of functional proteins by using ribosome display. Proc. Natl. Acad. Sci. USA 94, 4937–4942.

    Article  PubMed  CAS  Google Scholar 

  16. Sawasaki, T., Hasegawa, Y., Tsuchimochi, M., et al. (2002) A bilayer cell-free protein synthesis system for high-throughput screening of gene products. FEBS Lett. 514, 102–105.

    Article  PubMed  CAS  Google Scholar 

  17. Spirin, A. S., Baranov, V. I., Ryabova, L. A., Ovodov, S. Y., and Alakhov, Y. B. (1988) A continuous cell-free translation system capable of producing polypeptides in high yield. Science 242, 1162–1164.

    Article  PubMed  CAS  Google Scholar 

  18. Endo, Y. and Sawasaki, T. (2004) High-throughput, genome-scale protein production method based on the wheat germ cell-free expression system. J. Struct. Funct. Genomics 5(1–2), 45–57.

    Article  PubMed  CAS  Google Scholar 

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© 2005 Humana Press Inc.

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Sawasaki, T. et al. (2005). The Wheat Germ Cell-Free Expression System. In: Zanders, E.D. (eds) Chemical Genomics. Methods in Molecular Biology™, vol 310. Humana Press. https://doi.org/10.1007/978-1-59259-948-6_10

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  • DOI: https://doi.org/10.1007/978-1-59259-948-6_10

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-399-2

  • Online ISBN: 978-1-59259-948-6

  • eBook Packages: Springer Protocols

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