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Screening arrayed libraries with DNA and protein baits to identify interacting proteins

A simple straightforward procedure for high-throughput screenings of arrayed libraries with DNA or protein baits is described.

CBGP (UPM-INIA) researchers, of the group “Studying gibberellin signalling to improve seed germination and plant growth under stress”, provide detailed information and guidelines to improve the detection of binary interactions involving DNA and proteins. Most cellular functions in living organisms rely on intricate molecular interactions. In particular, interactions between nucleic acids and proteins 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 gene 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 labour 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.


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Original Paper:

Sánchez-Montesino, R; Oñate-Sánchez, L. 2018. "Screening arrayed libraries with DNA and protein baits to identify interacting proteins", p. 131-149. In L. Oñate-Sánchez (ed.), Two-Hybrid Systems: Methods and Protocols. Springer New York, New York, NY. DOI: 10.1007/978-1-4939-7871-7_9".

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