Yeast Two-Hybrid Library Screening to study protein-protein interactions (CAT#: STEM-MB-0016-WXH)

Introduction

Library screening is the process of identification of the clones carrying the gene of interest. Screening relies on a unique property of a clone in a library. Screening can be based on detecting the DNA sequence of the cloned gene, detecting a protein that the gene encodes, or the use of linked DNA markers.<br />Two-hybrid screening (originally known as yeast two-hybrid system or Y2H) is a molecular biology technique used to discover protein–protein interactions (PPIs) and protein–DNA interactions by testing for physical interactions (such as binding) between two proteins or a single protein and a DNA molecule, respectively.<br />The premise behind the test is the activation of downstream reporter gene(s) by the binding of a transcription factor onto an upstream activating sequence (UAS). For two-hybrid screening, the transcription factor is split into two separate fragments, called the DNA-binding domain (DBD or often also abbreviated as BD) and activating domain (AD). The BD is the domain responsible for binding to the UAS and the AD is the domain responsible for the activation of transcription. The Y2H is thus a protein-fragment complementation assay.