Jeno Gyuris

Interaction Trap/Two‐Hybrid System to Identify Interacting Proteins

The yeast two‐hybrid method (or interaction trap) is a powerful technique for detecting protein interactions. The procedure is performed using transcriptional activation of a dual reporter system in yeast to identify interactions between a protein of interest (the bait protein) and the candidate proteins for interaction. The method can be used to screen a protein library for interactions with a bait protein or to test for association between proteins that are expected to interact based on prior evidence. Interaction mating facilitates the screening of a library with multiple bait proteins. Curr. Protoc. Cell Biol. 53:17.3.1‐17.3.35.

UNIT 19.2 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins

The parameters and experimental conditions for this important system are constantly undergoing improvement. This newest version includes expanded tables describing interaction trap components and additional libraries compatible with the interaction trap system. It also features a new protocol on performing a hunt by interaction mating. Some of the commercial vendors selling yeast two‐hybrid reagents recommend using interaction mating to perform a hunt, so this procedure should be of great interest.

UNIT 17.3 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins

This unit presents protocols designed to detect interacting proteins. Using yeast as a “test tube” and transcriptional activation of a reporter system, interacting proteins can be identified. The system can also be used to test complex formation for proteins for which there exists a reason to expect interaction.