Isolation and Analysis of Bacterial Ribosomes Through Sucrose Gradient Ultracentrifugation.

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ID: 76111
2020
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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.
Reference Key
dos-santos2020isolationmethods Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Dos Santos, Ricardo F;Arraiano, Cecília M;Andrade, José M;
Journal methods in molecular biology (clifton, nj)
Year 2020
DOI
10.1007/978-1-0716-0231-7_19
URL
Keywords
ribosome isolation ribosome profiles ribosomes sucrose gradient uv analysis ultracentrifugation

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