Analysis of Prevacuolar Compartment-Mediated Vacuolar Proteins Transport View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2014

AUTHORS

Caiji Gao , Yi Cai , Xiaohong Zhuang , Liwen Jiang

ABSTRACT

Transient expression using protoplasts is a quick and powerful tool for studying protein trafficking and subcellular localization in plant cells. Prevacuolar compartments (PVCs) or multivesicular bodies (MVBs) are intermediate compartments that mediate protein transport between late Golgi or trans-Golgi network (TGN) and vacuole. Both wortmannin treatment and ARA7(Q69L) expression can induce PVC homotypic fusion and PVC enlargement in plant cells. Here, we describe detailed protocols to use transient expression of protoplasts derived from Arabidopsis suspension culture cells for studying protein trafficking and localization. Using three GFP-tagged vacuolar cargo proteins and RFP-tagged PVC membrane marker as examples, we illustrate the major tools and methods, including wortmannin treatment, ARA7(Q69L) expression and immunoblot analysis, to analyze PVC-mediated vacuolar protein transport in plant cells. More... »

PAGES

119-29

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-4939-1420-3_9

DOI

http://dx.doi.org/10.1007/978-1-4939-1420-3_9

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1019973782

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/25117279


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