Yeast One- and Two-Hybrid High-Throughput Screenings Using Arrayed Libraries

Sánchez Montesino, Rocío and Oñate Sanchez, Luis (2017). Yeast One- and Two-Hybrid High-Throughput Screenings Using Arrayed Libraries. "Methods in Molecular Biology", v. 1629 ; pp. 47-65. ISSN 1064-3745. https://doi.org/10.1007/978-1-4939-7125-1_5.

Description

Title: Yeast One- and Two-Hybrid High-Throughput Screenings Using Arrayed Libraries
Author/s:
  • Sánchez Montesino, Rocío
  • Oñate Sanchez, Luis
Item Type: Article
Título de Revista/Publicación: Methods in Molecular Biology
Date: 2017
ISSN: 1064-3745
Volume: 1629
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: E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM)
Department: Otro
Creative Commons Licenses: Recognition - No derivative works - Non commercial

Full text

[img]
Preview
PDF - Requires a PDF viewer, such as GSview, Xpdf or Adobe Acrobat Reader
Download (1MB) | Preview

Abstract

Since their original description more than 25 years ago, the yeast one- and two-hybrid systems (Y1H/Y2H) have been used by many laboratories to detect DNA?protein (Y1H) and protein?protein interactions (Y2H). These systems use yeast cells (Saccharomyces cerevisiae) as a eukaryotic ?test tube? and are amenable for most labs in the world. The development of highly efficient cloning methods has fostered the generation of large collections of open reading frames (ORFs) for several organisms that have been used for yeast screenings. Here, we describe a simple mating based method for high-throughput screenings of arrayed ORF libraries with DNA (Y1H) or protein (Y2H) baits not requiring robotics. One person can easily carry out this protocol in approximately 10 h of labor spread over 5 days. It can also be scaled down to test one-to-one (few) interactions, scaled up (i.e., robotization) and is compatible with several library formats (i.e., 96, 384-well microtiter plates).

Funding Projects

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

More information

Item ID: 53014
DC Identifier: http://oa.upm.es/53014/
OAI Identifier: oai:oa.upm.es:53014
DOI: 10.1007/978-1-4939-7125-1_5
Official URL: https://link.springer.com/protocol/10.1007%2F978-1-4939-7125-1_5
Deposited by: Memoria Investigacion
Deposited on: 19 Nov 2018 12:36
Last Modified: 23 Apr 2019 13:12
  • Logo InvestigaM (UPM)
  • Logo GEOUP4
  • Logo Open Access
  • Open Access
  • Logo Sherpa/Romeo
    Check whether the anglo-saxon journal in which you have published an article allows you to also publish it under open access.
  • Logo Dulcinea
    Check whether the spanish journal in which you have published an article allows you to also publish it under open access.
  • Logo de Recolecta
  • Logo del Observatorio I+D+i UPM
  • Logo de OpenCourseWare UPM