Yeast Two-hybrid (Y2H) Library Construction to study protein-protein interactions (CAT#: STEM-MB-0013-WXH)

Introduction

The yeast library extracts the total RNA of the species, then isolates the mRNA, and obtains cDNA by reverse transcription. Afterwards, we connect these cDNAs to the AD vector to obtain an AD library, and then transfer it into yeast. In theory, the yeast library can express all proteins of the studied species.
The principle of Yeast two hybrid technology: Yeast transcription factor GAL4 contains two interacting but spatially separated domains. One is the DNA-binding domain (BD), and the other is the transcription activating domain (AD). When the two domains are close enough, the downstream reporter gene can be activated. Therefore, the two proteins to be studied can be linked to vectors containing AD and BD, and then transfected into yeast. If these two proteins interact with each other, it will bring AD and BD to meet spatially, thereby activating the downstream reporter gene.