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© Carmen Buchrieser, Marie-Christine Prevost
Legionella pneumophila et son flagelle, bactérie responsable de pneumopathie aigue grave. Bactérie de l'environnement , l'émergence récente de cette maladie s'explique par son affinité pour les systèmes modernes d'alimentation en eau comme les tours de refroidissement. Image colorisée.
Publication : Methods in molecular biology (Clifton, N.J.)

Co-immunoprecipitation: protein-RNA and protein-DNA interaction

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Methods in molecular biology (Clifton, N.J.) - 01 Jan 2013

Sahr T, Buchrieser C

Link to Pubmed [PMID] – 23150422

Methods Mol. Biol. 2013;954:583-93

Transcriptional and posttranscriptional regulators play a critical role in allowing a bacterium to adapt to the diverse environments and conditions it encounters. In order to characterize the role of these regulators the identification of their specific interaction partners is of utmost importance. Co-immunoprecipitation (IP) is based on antigen/antibody complex formation to purify a protein of interest from the rest of the samples together with its interaction partner. This method allows us to study direct interaction of a regulator with its specific binding partners like protein-RNA, protein-DNA, or protein-protein interactions. IP typically requires careful optimization and troubleshooting depending on the varying physicochemical characteristics of the protein of interest. In this chapter we present a starting point and the basic guidelines to obtain the best possible results from an IP experiment with subsequent use of new-generation sequencing techniques to detect mRNA or ncRNA targets (RIPseq) and protein-DNA interactions (ChIPseq).

http://www.ncbi.nlm.nih.gov/pubmed/23150422