Screening Arrayed Libraries with DNA and Protein Baits to Identify Interacting Proteins

Sánchez Montesino, Rocío and Oñate Sanchez, Luis (2018). Screening Arrayed Libraries with DNA and Protein Baits to Identify Interacting Proteins. "Methods in Molecular Biology", v. 1794 ; pp. 131-149. ISSN 1064-3745. https://doi.org/10.1007/978-1-4939-7871-7_9.

Description

Title: Screening Arrayed Libraries with DNA and Protein Baits to Identify Interacting Proteins
Author/s:
  • Sánchez Montesino, Rocío
  • Oñate Sanchez, Luis
Item Type: Article
Título de Revista/Publicación: Methods in Molecular Biology
Date: 2018
ISSN: 1064-3745
Volume: 1794
Subjects:
Freetext Keywords: Arrayed libraries; DNA–protein interaction; High-throughput; One-hybrid system; Open reading frame; Protein–protein interaction; Transcription factors; Two-hybrid system; Yeast
Faculty: Centro de Investigación en Biotecnología y Genómica de Plantas (CBGP) (UPM)
Department: Otro
Creative Commons Licenses: Recognition - No derivative works - Non commercial

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Abstract

Molecular interactions are an integral part of the regulatory mechanisms controlling gene expression. The yeast one- and two-hybrid systems (Y1H/Y2H) have been widely used by many laboratories to detect DNA?protein (Y1H) and protein?protein interactions (Y2H). The development of efficient cloning systems have promoted the generation of large open reading frame (ORF) clone collections (libraries) for several organisms. Functional analyses of such large collections require the establishment of adequate protocols. Here, we describe a simple straightforward procedure for high-throughput screenings of arrayed libraries with DNA or protein baits that can be carried out by one person with minimal labor and not requiring robotics. The protocol can also be scaled up or down and is compatible with several library formats. Procedures to make yeast stocks for long-term storage (tube and microplate formats) are also provided.

Funding Projects

TypeCodeAcronymLeaderTitle
Government of SpainBIO2013-46076-RUnspecifiedUnspecifiedUnspecified
Government of SpainBIO2016-77840-RUnspecifiedUnspecifiedUnspecified

More information

Item ID: 52992
DC Identifier: http://oa.upm.es/52992/
OAI Identifier: oai:oa.upm.es:52992
DOI: 10.1007/978-1-4939-7871-7_9
Official URL: https://link.springer.com/protocol/10.1007/978-1-4939-7871-7_9
Deposited by: Memoria Investigacion
Deposited on: 30 Nov 2018 12:58
Last Modified: 01 Jul 2019 22:30
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