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Visualizing Secretory Cargo Transport in Budding Yeast.


ABSTRACT: Budding yeast is an excellent model organism for studying the dynamics of the Golgi apparatus. To characterize Golgi function, it is important to visualize secretory cargo as it traverses the secretory pathway. We describe a recently developed approach that generates fluorescent protein aggregates in the lumen of the yeast endoplasmic reticulum and allows the fluorescent cargo to be solubilized for transport through the Golgi by addition of a small-molecule ligand. We further describe how to generate a yeast strain expressing the regulatable secretory cargo, and we provide protocols for visualizing the cargo by 4D confocal microscopy and immunoblotting. © 2018 by John Wiley & Sons, Inc.

SUBMITTER: Casler JC 

PROVIDER: S-EPMC6506369 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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Visualizing Secretory Cargo Transport in Budding Yeast.

Casler Jason C JC   Glick Benjamin S BS  

Current protocols in cell biology 20181110 1


Budding yeast is an excellent model organism for studying the dynamics of the Golgi apparatus. To characterize Golgi function, it is important to visualize secretory cargo as it traverses the secretory pathway. We describe a recently developed approach that generates fluorescent protein aggregates in the lumen of the yeast endoplasmic reticulum and allows the fluorescent cargo to be solubilized for transport through the Golgi by addition of a small-molecule ligand. We further describe how to gen  ...[more]

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