Abstract
The wheat germ cell-free protein synthesis system has been used as a eukaryotic protein production system since it was first reported in 1964. Although initially the productivity of this system was not very high, it has now become one of the most versatile protein production systems, thanks to the enhancements made by several groups. In this chapter, we report a protein production method for plant receptor kinases using the wheat cell-free system. We describe a method for the preparation of a cell-free extract from wheat germ, the split-primer PCR method for preparation of transcription templates, and the bilayer cell-free protein synthesis method.
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References
Marcus A, Feely J (1964) Activation of protein synthesis in imbibition phase of seed germination. Proc Natl Acad Sci U S A 51:1075–1079
Spirin AS, Baranov VI, Ryabova LA, Ovodov SY, Alakhov YB (1988) A continuous cell-free translation system capable of producing polypeptide in high yield. Science 242:1162–1164
Kawarasaki Y, Kawai T, Nakano H, Yamane T (1995) A long-lived batch reaction system of cell-free protein synthesis. Anal Biochem 226:320–324
Nakano H, Tanaka T, Kawarasaki Y, Yamane T (1996) Highly productive cell-free protein synthesis system using condensed wheat-germ extract. J Biotechnol 46:275–282
Madin K, Sawasaki T, Ogasawara T, 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 U S A 97:559–564
Kamura N, Sawasaki T, Kasahara Y, Takai K, Endo Y (2005) Selection of 5′-untranslated sequences that enhance initiation of translation of a cell-free protein synthesis system from wheat embryos. Bioorg Med Chem Lett 15:5402–5406
Sawasaki T, Ogasawara T, Morishita R, Endo Y (2002) A cell-free protein synthesis system for high-throughput proteomics. Proc Natl Acad Sci U S A 99:14652–14657
Sawasaki T, Hasegawa Y, Tsuchimochi M, Kamura N, Ogasawara T, Kuroita T, Endo Y (2002) A bilayer cell-free protein synthesis system for high-throughput screening of gene products. FEBS Lett 514:102–105
Vinarov DA, Lytle BL, Peterson FC, Tyler EM, Volkman BF, Markley JL (2004) Cell-free protein production and labeling protocol for NMR-based structural proteomics. Nat Methods 1:149–153
Nemoto K, Takemori N, Seki M, Shinozaki K, Sawasaki T (2015) Members of the plant CRK superfamily are capable of trans- and autophosphorylation of tyrosine residues. J Biol Chem 290:16665–16677
Gish LA, Clark SE (2011) The RLK/Pelle family of kinases. Plant J 66:117–127
Morillo SA, Tax FE (2006) Functional analysis of receptor-like kinases in monocots and dicots. Curr Opin Plant Biol 9:460–469
The Arabidopsis Genome Initiative (2000) Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 408:796–815
Dardick C, Chen J, Richter T, Ouyang S, Ronald P (2007) The rice kinase database. A phylogenetic database for the rice kinome. Plant Physiol 143:579–586
Shiu SH, Bleecker AB (2003) Expansion of the receptor-like kinase/Pelle gene family and receptor-like proteins in Arabidopsis. Plant Physiol 132:530–543
Nemoto K, Seto T, Takahashi H, Nozawa A, Seki M, Shinozaki K, Endo Y, Sawasaki T (2011) Autophosphorylation profiling of Arabidopsis protein kinases using the cell-free system. Phytochemistry 72:1136–1144
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Nozawa, A., Nemoto, K., Nomura, S., Yamanaka, S., Kido, K., Sawasaki, T. (2017). Cell-Free Synthesis of Plant Receptor Kinases. In: Aalen, R. (eds) Plant Receptor Kinases. Methods in Molecular Biology, vol 1621. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7063-6_4
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DOI: https://doi.org/10.1007/978-1-4939-7063-6_4
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7063-6
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