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Yeast Two-Hybrid (Y2H) is an in vivo protein-protein interaction screening technology based on the modular structure of transcription factors. The system splits the GAL4 transcription factor into a DNA-Binding Domain (BD) and a Transcription Activation Domain (AD), which are fused to a Bait protein and a Prey protein, respectively; when the Bait and Prey physically interact within the yeast nucleus, the BD and AD are brought into proximity and reconstitute a functional transcription factor, activating downstream reporter genes (e.g., HIS3, ADE2, LacZ). Protein interaction is thus determined through auxotrophic complementation or colorimetric assays. This technique requires no protein purification and enables high-throughput screening of unknown interaction partners at the whole-genome cDNA library scale, serving as a classical tool for discovering novel protein-protein interactions.