Isolation and analysis of bacterial ribosomes through sucrose gradient ultracentrifugation

Ricardo F. dos Santos, Cátia Bárria, Cecília M. Arraiano, José M. Andrade

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Citation (Scopus)

Abstract

Ribosomes are large macromolecular complexes responsible for the translation process. During the course of ribosome biogenesis and protein synthesis, extra-ribosomal factors interact with the ribosome or its subunits to assist in these vital processes. Here we describe a method to isolate and analyze not only bacterial ribosomes but also their associated factors, providing insights into translation regulation. This detailed protocol allows the separation and monitoring of the ribosomal species and their interacting partners along a sucrose density gradient. Simultaneously, fractionation of the gradient allows for the recovery of 70S ribosomes and its subunits enabling a wide range of downstream applications. This protocol can be easily adapted to ribosome-related studies in other species or for separating other macromolecular complexes.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages299-310
Number of pages12
DOIs
Publication statusPublished - 2020

Publication series

NameMethods in Molecular Biology
Volume2106
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Ribosome isolation
  • Ribosome profiles
  • Ribosomes
  • Sucrose gradient
  • Ultracentrifugation
  • UV analysis

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