Mapping Plastid Transcript Population by Circular Reverse Transcription Polymerase Chain Reaction

Florence Courtois; Livia Merendino

doi:10.1007/978-1-4939-8654-5_18

Mapping Plastid Transcript Population by Circular Reverse Transcription Polymerase Chain Reaction

Methods Mol Biol. 2018:1829:273-278. doi: 10.1007/978-1-4939-8654-5_18.

Authors

Florence Courtois¹, Livia Merendino^{2

3}

Affiliations

¹ Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Institut National Recherche Agronomique, Commissariat à l'Energie Atomique et aux Energies Alternatives, CEA Grenoble, UMR5168, Université Grenoble Alpes, Grenoble, France. florence.courtois@cea.fr.
² Centre National de la Recherche Scientifique, Institute of Plant Sciences Paris Saclay, Institut National de la Recherche Agronomique, Université Paris-Sud, Université Evry, Université Paris-Saclay, Paris Diderot, Sorbonne Paris-Cité, Gif sur Yvette Cedex, France.
³ Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Institut National Recherche Agronomique, Commissariat à l'Energie Atomique et aux Energies Alternatives, CEA Grenoble, UMR5168, Université Grenoble Alpes, Grenoble, France.

PMID: 29987728
DOI: 10.1007/978-1-4939-8654-5_18

Abstract

During evolution of photosynthetic organisms, the genetic information provided by the internalized cyanobacteria has been transferred to the nucleus. The small genome kept by the chloroplast, the so-called plastome, displays a complex organization, comprising operons under the control of multiples promoters. In addition, polycistronic transcripts undergo multiple processing events, thus generating a complex population of mRNAs from a single gene. This chapter describes a method to investigate the diversity of the mRNA population from a single gene by circular RT-PCR. The protocol provided here allows for the simultaneous mapping of both 5' and 3' ends of the same RNA molecule.

Keywords: Chloroplast; Circular PCR; Operon; RNA end mapping; Transcript analysis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Gene Expression Profiling* / methods
Plastids / genetics*
RNA, Plant
Reverse Transcriptase Polymerase Chain Reaction* / methods
Transcriptome*
Workflow

Substances

RNA, Plant