诱导酿酒酵母中从头生成脂滴的方案。

Protocol to induce de novo lipid droplets in Saccharomyces cerevisiae.

机构信息

Lipid Metabolism Laboratory, Department of Biotechnology, All India Institute of Medical Sciences (AIIMS), New Delhi 110029, India.

出版信息

STAR Protoc. 2023 Dec 15;4(4):102732. doi: 10.1016/j.xpro.2023.102732. Epub 2023 Nov 22.

DOI:10.1016/j.xpro.2023.102732

PMID:37995191

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10687286/

Abstract

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.

摘要

内质网中脂滴（LD）生物发生的机制尚不完全清楚。在这里，我们提供了一个在酿酒酵母中诱导含有三酰基甘油（TAG）或甾醇酯（SE）的新形成 LD 的方案。我们通过进行基因敲除并引入半乳糖诱导启动子来生成条件性酵母突变体，从而描述了产生含有 TAG 或 SE 的 LD 的步骤。我们详细介绍了通过荧光显微镜可视化新形成的液滴来生成荧光 LD 标记蛋白的策略。