Yeast Two Hybird System, DNA Binding Domain, Activation Domain, Gene Lying, Hybird Gene

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Home >> Biotechnology and Genomics >> Methods and Uses of Genomics and Proteomics Research >> Yeast Two Hybird System

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Yeast two-hybrid system.

This system is based on the knowledge that a transcription factor generally consists of a DNA binding domain (DBD), and an activation domain (AD), the former helping in DNA binding and the later facilitating activation of a gene lying downstream. If one wants to study interaction between two proteins X and Y, two hybrid gene constructs are prepared, one having a hybrid gene coding for fusion protein DBD + X along with a reporter gene and the other coding for the fusion protein AD + Y. Yeast cells are co-transformed with both hybrid constructs, so that if X and Y interact physically, this will bring DBD and AD in close proximity, thus permitting expression of reporter, gene.

In contrast, if X and Y do not interact, AD will not be available at DBD site and the reporter gene will not be expressed. This yeast two-hybrid- system is widely used for study of interaction between two specific proteins. The technique has been successfully used in preparing protein interaction maps for Helicobacter pylori (a bacterium) showing 1200 protein-protein interactions involvin about half of the proteins in the proteome of this organism and for S. cereviseae (budding yeast) showing 2240 protein-protein interactions.

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