UMMS Affiliation

Program in Gene Function and Expression; Program in Molecular Medicine

Publication Date

1999-11-24

Document Type

Article

Subjects

DNA, Fungal; Fungal Proteins; Gene Library; Genes, Fungal; Saccharomyces cerevisiae; Selection (Genetics); Trans-Activators; *Two-Hybrid System Techniques

Disciplines

Genetics and Genomics

Abstract

Large-scale sequencing projects have predicted high numbers of gene products for which no functional information is yet available. Hence, large-scale projects, such as gene knockouts, gene expression profiles, and protein-interaction mapping, are currently under way to initiate the understanding of the function of these gene products. The high-throughput strategies that are currently being developed to generate protein-interaction maps include automated versions of the yeast two-hybrid system. These strategies rely on the large-scale construction of DNA-binding domain/protein-of-interest hybrid constructs (DB-X baits). An inherent problem of large-scale two-hybrid systems is that a high percentage of cloned sequences encode polypeptides that, when fused to DB, can activate transcription in the absence of any two-hybrid-interacting partner protein. Here, we describe and validate a genetic strategy that efficiently eliminates such self-activator baits prior to screening procedures. The strategy is based on a negative-growth selection and is compatible with high-throughput settings.

DOI of Published Version

10.1101/gr.9.11.1128

Source

Genome Res. 1999 Nov;9(11):1128-34. Link to article on publisher's site

Journal/Book/Conference Title

Genome research

Comments

At the time of publication, Albertha J. Marian Walhout was not yet affiliated with the University of Massachusetts Medical School.

Related Resources

Link to Article in PubMed

PubMed ID

10568752

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