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Bacterial Regulatory RNA pp 351–370Cite as

Identification of Small RNA–Protein Partners in Plant Symbiotic Bacteria

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1737))

Abstract

The identification of the protein partners of bacterial small noncoding RNAs (sRNAs) is essential to understand the mechanistic principles and functions of riboregulation in prokaryotic cells. Here, we describe an optimized affinity chromatography protocol that enables purification of in vivo formed sRNA–protein complexes in Sinorhizobium meliloti, a genetically tractable nitrogen-fixing plant symbiotic bacterium. The procedure requires the tagging of the desired sRNA with the MS2 aptamer, which is affinity-captured by the MS2-MBP protein conjugated to an amylose resin. As proof of principle, we show recovery of the RNA chaperone Hfq associated to the strictly Hfq-dependent AbcR2 trans-sRNA. This method can be applied for the investigation of sRNA–protein interactions on a broad range of genetically tractable α-proteobacteria.

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Authors and Affiliations

  1. Grupo de Ecología Genética de la Rizosfera, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas (CSIC), Granada, Spain

    Marta Robledo, Natalia I. García-Tomsig & José I. Jiménez-Zurdo

  2. Faculty of Health and Medical Sciences, Department of Microbial and Cellular Sciences, School of Biosciences and Medicine, University of Surrey, Guildford, UK

    Ana M. Matia-González

Authors
  1. Marta Robledo
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  2. Ana M. Matia-González
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  3. Natalia I. García-Tomsig
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  4. José I. Jiménez-Zurdo
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Corresponding author

Correspondence to José I. Jiménez-Zurdo .

Editor information

Editors and Affiliations

  1. Université Paris Diderot & LLB/CEA/CNRS, Gif-sur-Yvette, France

    Véronique Arluison

  2. Laboratorio de Bioquímica, Microbiología e Interacciones, Biológicas en el Suelo, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes-CONICET, Bernal, Argentina

    Claudio Valverde

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Robledo, M., Matia-González, A.M., García-Tomsig, N.I., Jiménez-Zurdo, J.I. (2018). Identification of Small RNA–Protein Partners in Plant Symbiotic Bacteria. In: Arluison, V., Valverde, C. (eds) Bacterial Regulatory RNA. Methods in Molecular Biology, vol 1737. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7634-8_20

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  • DOI: https://doi.org/10.1007/978-1-4939-7634-8_20

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7633-1

  • Online ISBN: 978-1-4939-7634-8

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