TwoHybridExperimenten
Two-Hybrid Experiments are a set of molecular biology methods designed to detect protein–protein interactions in vivo by reconstituting a functional transcription factor from two separate domains. The most widely used version is the yeast two-hybrid (Y2H) assay, introduced by Fields and Song in 1989, which popularized the concept. The principle is that a protein of interest, called the bait, is fused to a DNA-binding domain, while a second protein, called the prey, is fused to a transcriptional activation domain. If bait and prey interact, the two domains are brought together and drive expression of a reporter gene in the host cell.
Common reporters include HIS3, ADE2, or lacZ in yeast, providing growth selection or colorimetric readouts. Yeast
Typical workflow involves cloning the two proteins into compatible bait and prey vectors, introducing them into
Limitations include false positives caused by auto-activation or non-specific interactions and false negatives due to improper