Abstract
Ribosome profiling (also known as Ribo-seq) provides a genome-wide, high-resolution, and quantitative accounting of mRNA segments that are occupied by ribosomes in vivo. The method has been used to address numerous questions in bacteria, yeast, and metazoa, but its application to questions in plant biology is just beginning. This chapter provides a detailed protocol for profiling ribosomes in plant leaf tissue. The method was developed and optimized with maize, but it has been used successfully with Arabidopsis and tobacco as well. The method captures ribosome footprints from the chloroplast and cytosol in the same preparation, but it is not optimal for detecting the footprints of mitochondrial ribosomes. The protocol is robust and simpler than many of the methods reported previously for ribosome profiling in plants.
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Acknowledgments
We are grateful to Roz Williams-Carrier (University of Oregon) for helpful input during development of this procedure, and Indrajit Kumar (Danforth Center) for helpful discussions on data analysis.
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Chotewutmontri, P., Stiffler, N., Watkins, K.P., Barkan, A. (2018). Ribosome Profiling in Maize. In: Lagrimini, L. (eds) Maize. Methods in Molecular Biology, vol 1676. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7315-6_10
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DOI: https://doi.org/10.1007/978-1-4939-7315-6_10
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