Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae

Abstract

Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• cell Biology
• model Organisms
• Molecular Biology
• protein Biochemistry