Yeast One-hybrid Library Construction to study DNA-protein interactions (CAT#: STEM-MB-0017-WXH)

Introduction

Yeast one-hybrid technology is a technical method for studying the interaction between proteins and specific DNA sequences. The initiation of transcription of eukaryotic genes requires the participation of transcription factors, which usually consist of a DNA-specific binding domain and one or more activation domains that interact with other regulatory proteins, namely the DNA-binding domain (DNA-binding domain, BD) and transcriptional activation domain (activationdomain, AD). The yeast GAL4 protein used in the yeast one-hybrid system is a typical transcription factor. The DNA-binding domain of GAL4 is near the carboxy terminus and contains several zinc finger structures that activate the upstream activation site (UAS) of yeast galactosidase. And the transcription activation domain can interact with RNA polymerase or transcription factor TFIID to increase the activity of RNA polymerase. In this process, the DNA-binding domain and the transcriptional activation domain can function completely independently. Accordingly, we can replace the DNA-binding domain of GAL4 with a library protein-coding gene. As long as the expressed protein can interact with the target gene, it can also activate RNA polymerase through the transcription activation domain and initiate the transcription of the downstream reporter gene.