Transcriptional regulation improves the throughput of three-hybrid counter selections in Saccharomyces cerevisiae.

ABSTRACT: The yeast three-hybrid (Y3H) assay expands the fields of drug discovery and protein engineering by enabling the search of large variant libraries for targets that do not inherently produce a distinct, measurable phenotype. The Y3H assay links the DNA-binding and activation domains of a transcription factor via a chemically synthesized heterodimeric small molecule, thereby activating a downstream reporter gene. Although the Y3H assay has been successfully applied as a positive selection to discover novel drug targets and to evolve proteins with improved functions, its expansion into applications requiring a high-throughput, versatile selection against transcriptional activation has been hindered by its limited dynamic range as a counter selection. Here, we describe the development of a seco

SUBMITTER: Harton MD 

PROVIDER: S-EPMC8896811 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature