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In Vivo Analysis of RNA Editing in Plastids

  • Stephanie Ruf
  • Ralph BockEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 718)

Abstract

mRNA editing in plastids (chloroplasts) of higher plants proceeds by cytidine-to-uridine conversion at highly specific sites. Editing sites are recognized by the interplay of cis-acting elements at the RNA level and site-specific trans-acting protein factors that are believed to bind to the cis-elements in a sequence-specific manner. The C-to-U editing enzyme, a presumptive cytidine deaminase acting on polynucleotides, is still unknown. The development of methods for the stable genetic transformation of the plastid genome in higher plants has facilitated the analysis of RNA editing in vivo. Plastid transformation has been extensively used to define the sequence requirements for editing site selection and to address questions about editing site evolution. This chapter describes the basic methods involved in the generation and analysis of plants with transgenic chloroplast genomes and summarizes the applications of plastid transformation in editing research.

Key words

RNA editing Plastid Chloroplast Nicotiana tabacum Plastid transformation Biolistic transformation Particle gun cis-Acting element Evolution 

Notes

Acknowledgments

Work on RNA editing and plastid transformation in the authors’ laboratory is supported by the Max Planck Society and by grants from the Deutsche Forschungsgemeinschaft (DFG), the Bundesministerium für Bildung und Forschung (BMBF), and the European Union (Framework Programs 6 and 7).

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Authors and Affiliations

  1. 1.Max-Planck-Institut für Molekulare PflanzenphysiologiePotsdam-GolmGermany
  2. 2.Max Planck Institute of Molecular Plant PhysiologyPotsdam-GolmGermany

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