第四部分+4-5+目基因获得-酵母双杂交.pdfVIP

第四部分 目的基因的获得 五.酵母双杂交系统分离目的基因 酵母双杂交系统 (Yeast Two-hybrid System) 用途： 快速筛选与一种已知蛋白结合的目的蛋 白，发现新基因 发现新基因 验证 已知蛋白间的相互作用及作用强度 验证 转录因子的组件结构特性 （a)酵母转录因子GCN4的结构域；（b）哺乳动物糖皮质激素受体蛋 白质的结构域。负责特定基因对糖皮质激素反应的转录因子 细胞因子Oct1－1对基因转录的激活作用 通过与具有激活域的单纯疱疹病毒VP16的作用激活基因转录 Yeast two-hybrid system: a genetic assay for detecting protein-protein interactions Regulation of gene expression in yeast transcription activator activation domain transcription machinery DNA binding domain UAS gene (upstream activation sequence) Yeast two-hybrid system: a genetic assay for detecting protein-protein interactions Regulation of gene expression in yeast Do these transcription activator two protein activation domain transcription machinery bind? DNA binding domain UAS gene Y (upstream activation sequence) X We start with yeast possessing a reporter gene (a gene making a product that is easy to detect). And now we introduce genes coding for two hybrid proteins reporter gene His, β-gal Yeast two-hybrid system: a genetic assay for detecting protein-protein interactions Regulation of gene expression in yeast transcription activator