Abstract
In vitro functional studies of eukaryotic kinases are often constrained by the availability of pure and enzymatically active kinase of interest. Though numerous proteins have been synthesized by cell-based systems, in vivo production of properly folded, eukaryotic proteins remains a challenging task. Current wheat-germ-based cell-free in vitro translation systems present a plausible alternative for protein synthesis since majority of eukaryotic proteins could be obtained in their native folded form with general protocols. The use of special in vitro translation vectors with ligation-independent cloning sites and cleavable affinity tags eliminates further bottlenecks of the protein producing procedure and makes this system a reasonable method for simultaneous generation of active kinases.
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This work was supported by Hungarian Scientific Research Fund and National Office for Research and Technology grant K69187 and K68160.
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Sonkoly, B., Bardóczy, V., Mészáros, T. (2011). Expression and Purification of Active Protein Kinases from Wheat Germ Extracts. In: Dissmeyer, N., Schnittger, A. (eds) Plant Kinases. Methods in Molecular Biology, vol 779. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-61779-264-9_3
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DOI: https://doi.org/10.1007/978-1-61779-264-9_3
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Publisher Name: Humana, Totowa, NJ
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Online ISBN: 978-1-61779-264-9
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