Abstract
Orderly progression of the eukaryotic cell cycle is governed by a coordinated response to intrinsic and extracellular cues through activation of cyclin-dependent kinases (CDKs). It is therefore important to verify the kinase activity of distinct types of CDKs during the cell cycle. The immunoprecipitation-coupled kinase assay is a useful procedure to evaluate CDK activity in vivo. Although a specific antibody is usually required for immunoprecipitation, transgenic plant cells expressing tag- or marker protein-fused CDKs are also suitable for this purpose. In addition, the baculovirus expression system is a valuable tool for analyzing CDK activity in vitro, because activation of CDKs is regulated by posttranscriptional modification systems that are active in the insect host cells.
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Acknowledgments
The authors wish to thank Drs. Ko Kato, Kazuya Yoshida, and Atsuhiko Shinmyo for their helpful discussions and suggestions. The authors also wish to thank Dr. Ian Smith for critical reading of the manuscript. This research was supported by a Grant-in-Aid for Scientific Research from the Japanese Ministry of Education, Culture, Sports, Science and Technology and by a Grant-in-Aid for Creative Scientific Research.
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Harashima, H., Sekine, M. (2011). Measurement of Plant Cyclin-Dependent Kinase Activity Using Immunoprecipitation-Coupled and Affinity Purification-Based Kinase Assays and the Baculovirus Expression System. 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_4
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DOI: https://doi.org/10.1007/978-1-61779-264-9_4
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