双杂交筛选
蛋白质表达
酵母
表达式(计算机科学)
计算生物学
控制(管理)
生物
合成生物学
计算机科学
遗传学
基因
程序设计语言
人工智能
作者
Erez Feuer,Gil Zimran,Michal Shpilman,Heng Song
标识
DOI:10.1021/acssynbio.2c00192
摘要
The yeast two-hybrid (Y2H) assay is widely used for protein-protein interaction characterization due to its simplicity and accessibility. However, it may mask changes in affinity caused by mutations or ligand activation due to signal saturation. To overcome this drawback, we modified the Y2H system to have tunable protein expression by introducing a fluorescent reporter and a pair of synthetic inducible transcription factors to regulate the expression of interacting components. We found that the application of inducers allowed us to adjust the concentrations of interacting proteins to avoid saturation and observe interactions otherwise masked in the canonical Y2H assay, such as the abscisic acid-mediated increase in affinity of monomeric abscisic acid receptors to the coreceptor. When applied in future studies, our modified system may provide a more accurate characterization of protein-protein interactions.
科研通智能强力驱动
Strongly Powered by AbleSci AI