Generating Yeast Two-Hybrid Bait Strains
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Updated: Feb 8, 2026

Modified Yeast-Two-Hybrid System to Identify Proteins Interacting with the Growth Factor Progranulin
Published on: January 17, 2012
Abstract:
Generating DNA-binding domain (DB)-bait strains for Gateway-compatible yeast two-hybrid (Y2H) screens involves three steps. The first is to generate an Entry clone containing a DNA fragment encoding the protein of interest (e.g., an open reading frame, ORF). The second is to transfer this DNA fragment from the Entry clone to the Y2H Destination vector, pDEST32. The final step is to transform this construct into the Y2H yeast strain, MaV103. This protocol takes 24-37 d plus sequence confirmation, if necessary, to complete.
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