Abstract
To date a number of cereal genomes are fully sequenced and more are near completion. The information within these genomes will be of most use to scientists when every gene has been functionally characterized leading to the complete annotation of these genomes. This chapter describes how functional characterization of plant proteins can be achieved via in vitro or in vivo methods. The first section of this chapter describes the use of Escherichia coli as a host for expression of plant genes, followed by purification and in vitro characterization of the resultant enzyme. The second section of this chapter details the methods involved in transient gene expression in Zea mays leaf protoplasts for in vivo functional characterization of protein localization.
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Acknowledgements
I would like to thank Dr. Maria Shumskaya for her guidance in the procedures of protoplast isolation and transformation. Many thanks to Dr. Abby Cuttriss and Dr. Rémi Zallot for their comments and thanks to Prof. Eleanore Wurtzel and Dr. Maria Shumskaya for use of their protoplast images.
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Bradbury, L.M.T. (2014). Functional Analysis by Protein Biochemistry. In: Henry, R., Furtado, A. (eds) Cereal Genomics. Methods in Molecular Biology, vol 1099. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-715-0_13
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DOI: https://doi.org/10.1007/978-1-62703-715-0_13
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