酵母双杂交技术鉴定 PDZ 相互作用。

Identification of PDZ Interactions by Yeast Two-Hybrid Technique.

机构信息

Centre de Recherche en Cancerologie de Marseille (CRCM), Equipe Zimmermann labellisée Ligue 2018 - 2019, Aix-Marseille Université, Inserm, CNRS, Institut Paoli-Calmettes, Marseille, France.

Department of Human Genetics, K. U. Leuven, Leuven, Belgium.

出版信息

Methods Mol Biol. 2021;2256:1-15. doi: 10.1007/978-1-0716-1166-1_1.

DOI:10.1007/978-1-0716-1166-1_1

PMID:34014513

Abstract

The yeast two-hybrid technique is a powerful method to detect direct protein-protein interactions. Due to its accessibility, speed, and versatility, this technique is easy to set up in any laboratory and suitable for small and large scale screenings. Here we describe the implementation of an array-based screening that allows for the probing of the entire human PDZ ORFeome (or hPDZome) by yeast two-hybrid technique. With this approach, one can rapidly identify the PDZ domains that are able to interact (up to K in the high μmolar range) with any candidate protein among a panel of 266 individual clones, thereby comprehensively identifying its PDZ interactome.

摘要

酵母双杂交技术是一种检测直接蛋白质-蛋白质相互作用的强大方法。由于其易于使用、快速和多功能性，该技术在任何实验室都很容易建立，适用于小规模和大规模筛选。在这里，我们描述了一种基于阵列的筛选方法，该方法允许通过酵母双杂交技术探测整个人类 PDZ ORFeome（或 hPDZome）。通过这种方法，人们可以快速识别能够与面板中的任何候选蛋白相互作用（高达 K 在高 μ molar 范围内）的 PDZ 结构域，从而全面识别其 PDZ 相互作用组。