Abstract:
The two-hybrid system is a genetic method to identify proteins that interact with a specific target protein, which is expressed in yeast as a hybrid with a DNA-binding domain. Use of this method entails transforming yeast, both with this DNA-binding domain hybrid and with a library of activation domain hybrids, followed by screening for transcriptional activation of a reporter gene. In addition to proteins that interact with the target protein, a number of false positives are identified. We provide possible explanations for these false positives along with rapid assays to eliminate them.
摘要:
双杂交系统是一种遗传方法,用于鉴定与特定靶蛋白相互作用的蛋白质,在酵母中作为具有DNA结合域的杂合物表达。使用这种方法需要转化酵母,具有这种DNA结合域杂交体和激活域杂交体的文库,然后筛选报告基因的转录激活。除了与靶蛋白相互作用的蛋白质,识别出许多假阳性。我们提供了这些假阳性的可能解释以及消除它们的快速测定。