高通量蛋白质-蛋白质相互作用筛选的液基酵母双杂交池法。

High-Throughput Protein-Protein Interactions Screening Using Pool-Based Liquid Yeast Two-Hybrid Pipeline.

机构信息

Institute of Plant Sciences Paris-Saclay (IPS2), Université Paris-Saclay, Université Paris-Cité, CNRS, INRAE, Université Evry, Gif sur Yvette, France.

出版信息

Methods Mol Biol. 2023;2690:161-177. doi: 10.1007/978-1-0716-3327-4_16.

DOI:10.1007/978-1-0716-3327-4_16

PMID:37450147

Abstract

Because of its adaptability to high-throughput approaches and a low operating cost, the yeast two-hybrid (Y2H) assay remains the most widely used one for high-throughput protein-protein interactions (PPI) mapping experiments. Here we provide a detailed protocol for a liquid culture-based high-throughput binary protein-protein Y2H screen pipeline of a pool of 50 proteins used as baits against a collection of ~12,000 Arabidopsis proteins encoded by sequence-verified open reading frames (ORFs).

摘要

由于其适应高通量方法和低运营成本的特点，酵母双杂交（Y2H）测定仍然是最广泛用于高通量蛋白质 - 蛋白质相互作用（PPI）作图实验的方法。在这里，我们提供了一个详细的基于液体培养的高通量二元蛋白质 - 蛋白质 Y2H 筛选管道的协议，该管道使用 50 个作为诱饵的蛋白质池对大约 12000 个由经过序列验证的开放阅读框（ORFs）编码的拟南芥蛋白质进行筛选。