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Updated: Nov 30, 2025

An Assay for Quantifying Protein-RNA Binding in Bacteria
Published on: June 12, 2019
Protein Pulldown Assays to Monitor the Composition of the Bacterial RNA Degradosome
Steven W Hardwick1, Ben F Luisi1, Marilis V Marques2
1Department of Biochemistry, University of Cambridge, Cambridge, UK.
Abstract:
The method of co-immunoprecipitation (co-IP or pulldown) enables the identification of proteins interacting in macromolecular assemblies, through the purification of a key protein by affinity chromatography using specific antibodies immobilized on a matrix. The advantages of using epitope-tagged proteins include the ability to use commercially available antibodies for affinity purifications, and typically they do not disrupt the structure of the protein complexes. Here we describe the utilization of an epitope-tagged version of Caulobacter crescentus RNase E in order to determine the composition of the RNA degradosome under different growth conditions. Several proteins that interact with the RNA degradosome were identified.

