疟原虫红内期发育周期的全基因组核糖体分析。

Genome-Wide Ribosome Profiling of the Plasmodium falciparum Intraerythrocytic Developmental Cycle.

机构信息

Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA, USA.

Department of Biochemistry and Biophysics, University of California, San Francisco, CA, USA.

出版信息

Methods Mol Biol. 2021;2252:57-87. doi: 10.1007/978-1-0716-1150-0_3.

DOI:10.1007/978-1-0716-1150-0_3

PMID:33765271

Abstract

Monitoring whole-genome translation and mRNA ribosome occupancy in vivo using ribosome profiling has proven to be a powerful tool for discovery of gene expression regulation, mechanisms of translation, and new open reading frames, in a wide range of different cell types in different organisms. Here we describe its application to the malaria parasite, Plasmodium falciparum. We present methods for intact polysome purification from parasite cultures, polysome digestion, monosome purification, ribosome footprint nucleic acid extraction, and Illumina library preparation.

摘要

利用核糖体分析技术监测全基因组翻译和 mRNA 核糖体占据情况，已被证明是在不同生物的不同细胞类型中发现基因表达调控、翻译机制和新开放阅读框的有力工具。本文将介绍该技术在疟原虫（Plasmodium falciparum）中的应用。我们提供了从寄生虫培养物中纯化完整多核糖体、多核糖体消化、单核糖体纯化、核糖体足迹核酸提取和 Illumina 文库制备的方法。