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Trans-species Complementation Analysis to Study Function Conservation of Plant Endosomal Sorting Complex Required for Transport (ESCRT) Proteins

  • Francisca C. ReyesEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1209)

Abstract

ESCRT (Endosomal Sorting Complex Required for Transport) proteins are required for the sorting of biosynthetic and endocytic proteins at multivesicular bodies (MVBs). Here, I describe an assay to evaluate conservation of endosomal sorting functions of plant ESCRT proteins by trans-species complementation analysis. The assay is based on the imaging of a fluorescent biosynthetic MVB cargo, the carboxypeptidase S (CPS) fused to the green fluorescent protein GFP, in yeast ESCRT mutants expressing putative plant orthologues of the missing yeast ESCRT components.

Key words

Endosomal sorting Yeast ESCRT mutants MVB cargo Confocal microscopy 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Facultad Ciencias Biológicas, Centro de Biotecnología VegetalUniversidad Andrés BelloSantiagoChile

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