A genetic strategy to eliminate self-activator baits prior to high-throughput yeast two-hybrid screens
Document Type
Journal ArticlePublication Date
1999-11-24Keywords
DNA, FungalFungal Proteins
Gene Library
Genes, Fungal
Saccharomyces cerevisiae
Selection (Genetics)
Trans-Activators
*Two-Hybrid System Techniques
Genetics and Genomics
Metadata
Show full item recordAbstract
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.Source
Genome Res. 1999 Nov;9(11):1128-34. Link to article on publisher's siteDOI
10.1101/gr.9.11.1128Permanent Link to this Item
http://hdl.handle.net/20.500.14038/44116PubMed ID
10568752Notes
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 PubMedae974a485f413a2113503eed53cd6c53
10.1101/gr.9.11.1128