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Plant Immunity pp 181-194 | Cite as

Purification of Effector–Target Protein Complexes via Transient Expression in Nicotiana benthamiana

  • Joe Win
  • Sophien Kamoun
  • Alexandra M. E. JonesEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 712)

Abstract

Effectors of plant pathogens play important roles in not only pathogenesis but also plant immunity. Plant pathogens use these effectors to manipulate host cells for colonization, and their activities likely influence the evolution of plant immune responses. Analyses of genome sequences revealed that oomycete pathogens, such as Phytophthora spp., possess hundreds of RXLR effectors that are thought to be delivered into the host cells and hence function inside the cells by interacting with the host protein complexes. This article describes a co-immunoprecipitation protocol aimed at identifying putative target complexes of the effectors by transiently overexpressing the tagged effectors in planta. The identification of the eluted protein complexes was achieved by LC-MS/MS mass spectrometry and peptide spectrum matching.

Key words

Effectors Co-immunoprecipitation Oomycetes Phytophthora Mass spectrometry Target complexes Plant–microbe interaction 

Notes

Acknowledgments

We thank John Lindbo (UC Davis) for providing us with the TRBO vector, and Liliya Serazetdinova for technical assistance. This work was supported by the Gatsby Charitable Foundation.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Joe Win
    • 1
  • Sophien Kamoun
    • 1
  • Alexandra M. E. Jones
    • 1
    Email author
  1. 1.The Sainsbury LaboratoryJohn Innes CentreNorwichUK

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