Grow'n'Glow ACE1 Yeast Two-Hybrid System

The ACE1 Two-Hybrid System is a yeast two-hybrid system that efficiently identifies protein-protein interactions in vivo.

Yeast two-hybrid systems depend on the ability of specific separated DNA binding and transactivation domains to reconstitute an active transcription factor complex when interacting proteins are fused to these domains. Clones that express interacting proteins are consequently detected through the activity of reporter genes.

The ACE1 Two-Hybrid System identifies protein-protein interactions by combining the advantages of the copper-inducible transcription factor ACE1 and the copper resistance-mediating CUP1 gene, with the proven green fluorescence protein (GFP) as the reporter. With this combination, the interactions of two proteins fused to the DNA-binding and transactivation domains of ACE1 can be detected by growth of the cells on copper-containing media and the copper-dependent expression of the strongly-fluorescent GFPuv, which causes those colonies to fluoresce bright green under UV light.

The use of both a growth selection and a strong fluorescence signal creates a fast, highly sensitive and more reliable method of monitoring protein interactions in living cells. The copper concentration can be changed to select different protein-protein interaction strengths.