- A-Z
- Endocytobiosis and ...
- Volume 22 (2012)
- High-throughput ana...
- Autor(in)
- Seitenbereich
-
006-013
- Schlagwort(e)
-
protein-protein-interaction, transcription factor, interactome, yeast-two-hybrid, Arabidopsis, gene family
- Zusammenfsg.
-
The yeast two hybrid system is a powerful tool to determine interactions between two proteins in vivo. Expression of two proteins of interest fused either with a transcription activation domain (bait) or a DNA-binding domain (prey) of a eukaryotic transcription factor encoded in so-called bait and prey gene constructs in a single yeast cell enables the expression of distinct reporter genes, when both domains of the fused recombinant proteins meet in close proximity. This spatial arrangement of the two domains of a specific transcription factor is ensured when both proteins of interest interact with each other in the yeast strain. Suitable yeast cells can be generated by transforming yeast cells with both plasmids or alternatively by mating yeasts of opposite mating types each possessing either the bait or the prey construct. The latter technique enables the investigation of a plethora of different bait/prey combinations by mating appropriate yeast cells. Here we describe a high-throughput approach used for the investigation of the bilateral interactomic network among the subunits of an Eukaryotic transcription factor (TF) and their orthologs of different species. In contrast to their homologous representatives in heterotrophic organisms, plant TFs often are encoded by multigene families. The yeast two hybrid method enables a complete qualitative evaluation of potential interactions among multiple representatives of protein families.