Yeast Two Hybrid
Yeast Two Hybrid - Related Videos
05:20
Yeast Two-Hybrid Assay to Determine Protein Self-Association in Yeast Cells
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2025
This video demonstrates the yeast two-hybrid assay to detect specific protein self-interactions. In this assay, the protein of interest is tagged to two domains of a transcription factor that encodes the β-galactosidase enzyme. When the two proteins interact, this causes transcription and translation to form the enzyme, which is then detected using a biochemical assay.
07:56
Modified Yeast-Two-Hybrid System to Identify Proteins Interacting with the Growth Factor Progranulin
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Cited by 14 •
2012
We have modified the conventional yeast two-hybrid screening, an effective genetic tool in identifying protein interaction. This modification markedly shortens the process, reduces the workload, and most importantly, reduces the number of false positives. In addition, this approach is reproducible and reliable.
10:51
Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists
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Cited by 6 •
2013
Ketoconazole binds to and antagonizes Pregnane X Receptor (PXR) activation. Yeast high throughput screens of PXR mutants define a unique region for ketoconazole binding. This yeast-based genetic method discovers novel nuclear receptor interactions with ligands that associate with surface binding sites.
11:30
Unravelling the Function of a Bacterial Effector from a Non-cultivable Plant Pathogen Using a Yeast Two-hybrid Screen
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Cited by 8 •
2017
Bacterial effector proteins are important for establishing successful infections. This protocol describes the experimental identification of proteinaceous binding partners of a bacterial effector protein in its natural plant host. Identifying these effector interactions via yeast two-hybrid screens has become an important tool in unravelling molecular pathogenicity strategies.
04:12
Modified Yeast-One Hybrid Assay to Detect Heteromeric Protein Complex-DNA Interactions
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2025
In this video, we demonstrate the modified yeast one-hybrid assay, which detects multiple proteins interacting with target DNA. The transformed yeast cells contain the target DNA sequence upstream of the reporter gene. Additionally, they express transcription factor proteins. When the proteins form complexes and bind to the target DNA, the reporter gene is expressed and can be detected enzymatically.
FAQs
The assay converts a physical protein interaction into a transcriptional signal. One protein is linked to the DNA-binding domain and the other to the activation domain, which remain functionally separated when the proteins do not associate. Interaction reunites the domains, allowing transcription of the reporter gene and providing an indirect readout rather than a direct physical measurement.
The reporter gene is the detectable output of the assay. Its activation indicates that the two attached proteins brought the transcription-factor domains into functional proximity inside yeast. Consequently, researchers can score a candidate interaction through reporter activity, but the signal should be interpreted as evidence of interaction under assay conditions, not by itself as proof of behavior in the native cell.
Because the interaction is tested inside living yeast cells, the assay reports whether the two engineered protein constructs can produce a transcriptional response in a cellular setting rather than in an isolated mixture. However, yeast is not necessarily the proteins’ native cellular environment, so the result is informative for interaction studies but may need complementary validation.
Researchers select two proteins of interest, attach each to a different transcription-factor domain, and place the resulting constructs in living yeast. They then examine whether the reporter gene is activated. This workflow can test a suspected pair directly or support searches for binding partners, depending on the proteins selected for the experiment.
When one protein is used to search for previously unknown partners, the assay supports interaction discovery. When both proteins are selected in advance, reporter activation tests a suspected association. This distinction changes the question being asked, even though the underlying readout remains the same: transcription-factor domains become functionally reunited after the proteins interact.
Results from multiple protein-pair tests can help researchers identify binding partners and organize proteins into components of cellular signaling or regulatory networks. In this context, the assay provides interaction-level information that supports broader functional studies across biology. Interpreting those network relationships still benefits from complementary methods that assess interactions in native cellular contexts.