Our Matchmaker Gold Yeast One-Hybrid Library Screening System provides a simple and efficient method for identifying and characterizing novel protein-DNA interactions. All Matchmaker Gold Systems use Aureobasidin A resistance (AbA^r) as a stringent, highly selective, and easy-to-use reporter. This novel reporter produces very low screening backgrounds, since the Aureobasidin A antibiotic (AbA) efficiently kills yeast lacking AbA^r expression.

The Matchmaker Gold Yeast One-Hybrid Library Screening System

In the Matchmaker Gold Yeast One-Hybrid Library Screening System, tandem repeats of your DNA target/bait sequence are cloned into the pAbAi reporter vector. To generate your bait-specific reporter strain, the pAbAi vector is then integrated into the genome of Y1HGold yeast using homologous recombination. This strain provides a host for library screening.

One-Step library construction and screening

A cDNA library of potential DNA-binding proteins, which are ultimately expressed as fusions to the yeast GAL4 transcription activation domain (GAL4 AD prey proteins), is constructed directly in the Y1HGold[Bait-AbAi] reporter strain using SMART technology and homologous recombination. When a prey protein binds to the bait sequence, its associated GAL4 AD activates AbA^r expression, allowing the cell to grow on medium containing AbA. In library screens, the plasmids encoding the library-derived prey proteins can be easily rescued from the surviving yeast clones and subjected to further analysis.