Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae

The mechanisms underlying lipid droplet (LD) biogenesis in the endoplasmic reticulum are not completely understood. Here, we present a protocol for inducing de novo LDs containing either triacylglycerol (TAG) or sterol esters (SE) in Saccharomyces cerevisiae. We describe steps for generating conditi...

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Detalles Bibliográficos
Autores principales: Khatri, Amit, Choudhary, Vineet
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10687286/
https://www.ncbi.nlm.nih.gov/pubmed/37995191
http://dx.doi.org/10.1016/j.xpro.2023.102732
Descripción
Sumario:The mechanisms underlying lipid droplet (LD) biogenesis in the endoplasmic reticulum are not completely understood. Here, we present a protocol for inducing de novo LDs containing either triacylglycerol (TAG) or sterol esters (SE) in Saccharomyces cerevisiae. We describe steps for generating conditional yeast mutants by performing gene knockout and introducing galactose-inducible promoter to produce TAG- or SE-containing LDs. We detail the strategy to generate fluorescent LD marker protein to visualize newly formed droplets by fluorescence microscopy.

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