Yeast two-hybrid system is an important tool for studying protein function
The document describes the CytoTrap two-hybrid system, a method for detecting protein-protein interactions in the cytoplasm of yeast cells rather than the nucleus. This system overcomes limitations of conventional two-hybrid systems by utilizing a temperature-sensitive yeast strain (cdc25H) and the human Sos gene product to detect interactions that require posttranslational modifications or occur in the cytoplasm.
| Yeast Strain | cdc25H |
|---|---|
| Growth Temperature (Non-selective) | 25°C |
| Selection Temperature | 37°C |
| Carbon Source for Induction | Galactose |
| Carbon Source for Repression | Glucose |
It allows detection of interactions in the cytoplasm, enabling the study of proteins requiring posttranslational modifications or those that interact with transcriptional activators/inhibitors.
Positive interactions are selected by growing the yeast cells at 37°C, where only cells rescued by a protein-protein interaction can survive.
The pMyr vector is used for cDNA library construction, and the pSos vector is used for expressing the bait protein.