基于 SILAC 的果蝇胚胎定量蛋白质组学分析。

SILAC-Based Quantitative Proteomic Analysis of Drosophila Embryos.

机构信息

National Phenotypic Screening Centre, School of Life Sciences, University of Dundee, Dundee, UK.

Developmental Genetics Group, Institute for Biology, University of Kassel, Kassel, Germany.

出版信息

Methods Mol Biol. 2023;2603:187-198. doi: 10.1007/978-1-0716-2863-8_15.

DOI:10.1007/978-1-0716-2863-8_15

PMID:36370280

Abstract

The fruit fly Drosophila melanogaster represents a classic genetic model organism that is amenable to a plethora of comprehensive analyses including proteomics. SILAC-based quantitative proteomics is a powerful method to investigate the translational and posttranslational regulation ongoing in cells, tissues, organs, and whole organisms. Here we describe a protocol for routine SILAC labeling of Drosophila adults within one generation to produce embryos with a labeling efficiency of over 92%. In combination with genetic selection markers, this method permits the quantification of translational and posttranslational changes in embryos mutant for developmental and disease-related genes.

摘要

黑腹果蝇（Drosophila melanogaster）是一种经典的遗传模式生物，适用于包括蛋白质组学在内的多种综合分析。基于 SILAC 的定量蛋白质组学是一种强大的方法，可以研究细胞、组织、器官和整个生物体中正在进行的翻译和翻译后调控。在这里，我们描述了一种在一代内对成年果蝇进行常规 SILAC 标记的方案，可使标记效率超过 92%的胚胎。与遗传选择标记结合使用，该方法可定量检测发育和疾病相关基因的突变胚胎中的翻译和翻译后变化。