Protein Localization and Interaction Studies in Plants: Toward Defining Complete Proteomes by Visualization

Michael M. Goodin

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

7 Scopus citations

Abstract

Protein interaction and localization studies in plants are a fundamental component of achieving mechanistic understanding of virus:plant interactions at the systems level. Many such studies are conducted using transient expression assays in leaves of Nicotiana benthamiana, the most widely used experimental plant host in virology, examined by laser-scanning confocal microscopy. This chapter provides a workflow for protein interaction and localization experiments, with particular attention to the many control and supporting assays that may also need to be performed. Basic principles of microscopy are introduced to aid researchers in the early stages of adding imaging techniques to their experimental repertoire. Three major types of imaging-based experiments are discussed in detail: (i) protein localization using autofluorescent proteins, (ii) colocalization studies, and (iii) bimolecular fluorescence complementation, with emphasis on judicious interpretation of the data obtained from these approaches. In addition to establishing a general framework for protein localization experiments in plants, the need for proteome-scale localization projects is discussed, with emphasis on nuclear-localized proteins.

Original languageEnglish
Title of host publicationAdvances in Virus Research
EditorsMargaret Kielian, Thomas C. Mettenleiter, Marilyn J. Roossinck
Pages117-144
Number of pages28
DOIs
StatePublished - Jan 1 2018

Publication series

NameAdvances in Virus Research
Volume100
ISSN (Print)0065-3527
ISSN (Electronic)1557-8399

Bibliographical note

Publisher Copyright:
© 2018 Elsevier Inc.

Keywords

  • Colocalization
  • Confocal microscopy
  • Nicotiana benthamiana
  • Protein interaction

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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