通过环状逆转录聚合酶链反应绘制质体转录本群体图谱

Mapping Plastid Transcript Population by Circular Reverse Transcription Polymerase Chain Reaction.

作者信息

Courtois Florence, Merendino Livia

机构信息

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.

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.

出版信息

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

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

PMID:29987728

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.

摘要

在光合生物的进化过程中，被内化的蓝细菌所提供的遗传信息已转移至细胞核。叶绿体保留的小基因组，即所谓的质体基因组，呈现出复杂的组织形式，包含受多个启动子控制的操纵子。此外，多顺反子转录本会经历多种加工事件，从而从单个基因产生复杂的mRNA群体。本章描述了一种通过环状RT-PCR研究单个基因mRNA群体多样性的方法。此处提供的方案允许同时定位同一RNA分子的5'和3'末端。