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Identification of c-di-GMP-Responsive Riboswitches

descriptionPublicationkeyboard_double_arrow_right Part of book or chapter of book , Article 01 Jan 2017 France Publisher:Springer New York

Authors: Johann Peltier; Johann Peltier; Olga Soutourina; Olga Soutourina; Olga Soutourina;

doi: 10.1007/978-1-4939-7240-1_29

pmid: 28889309

Identification of c-di-GMP-Responsive Riboswitches

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Abstract

Bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) is an important signaling molecule for community behavior control, cell morphogenesis, and virulence in bacteria. In addition to protein effectors, this second messenger binds RNA molecules that act as riboswitches to control target gene expression. In this chapter, we describe a method for experimental validation of the functionality of c-di-GMP-responsive riboswitches and the analysis of c-di-GMP control of target gene expression by qRT-PCR and Northern blot. This procedure can be used for the studies of in silico-predicted riboswitch candidates, as well as a targeted experimental approach for exploring the data from next-generation sequencing. The examples on the analysis of type I and type II c-di-GMP-responsive riboswitches in Clostridium difficile are provided to illustrate the application of the method.

Country

France

Related Organizations

CEA LETI
France
University of Paris-Saclay
France
University of Paris
France
Institut Pasteur
France
Institute of Integrative Biology of the Cell
France

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Keywords

Transcription, Genetic, Clostridioides difficile, qRT-PCR, c-di-GMP, Northern blot, Blotting, Northern, Second Messenger Systems, Workflow, [SDV] Life Sciences [q-bio], RNA, Bacterial, Riboswitches, Riboswitch, Transcription Termination, Genetic, Nucleic Acid Conformation, Target gene, Premature termination of transcription, Cyclic GMP

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