Abstract
This chapter contains protocols for high-throughput protein production based on the cell-free system prepared from eukaryote wheat embryos.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Blaschke, U. K., Silberstein, J., and Muir, T. W. (2000) Protein engineering by expressed protein ligation. Methods Enzymol. 328, 478–496.
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.
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.
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.
Kurland, C. G. (1982) Translational accuracy in vitro. Cell 28, 201–202.
Pavlov, M. Y. and Ehrenberg, M. (1996) Mutants of EF-Tu defective in binding aminoacyl-tRNA. Arch. Biochem. Biophys. 328, 9–16.
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.
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.
Barbieri, L., Battelli, M. G., and Stirpe, F. (1993) Ribosome-inactivating proteins from plants. Biochim. Biophys. Acta 1154, 237–282.
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.
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.
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.
Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Gurevich, V. V. (1996) Use of bacteriophage RNA polymerase in RNA synthesis. Methods Enzymol. 275, 382–397.
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.
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.
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.
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
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
Download citation
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