Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77005 USA.
Department of Molecular and Cellular Medicine, Texas A&M University, College Station, TX, 77843 USA.
Biol Proced Online. 2011 Aug 15;13:6. doi: 10.1186/1480-9222-13-6.
Although yeast two-hybrid experiments are commonly used to identify protein interactions, the frequent occurrence of false negatives and false positives hampers data interpretation. Using both yeast one-hybrid and two-hybrid experiments, we have identified potential sources of these problems: the media preparation protocol and the source of the yeast nitrogen base may not only impact signal range but also effect whether a result appears positive or negative. While altering media preparation may optimize signal differences for individual experiments, media preparation must be reported in detail to replicate studies and accurately compare results from different experiments.
虽然酵母双杂交实验常用于鉴定蛋白质相互作用,但频繁出现的假阴性和假阳性结果会妨碍数据解释。通过使用酵母单杂交和双杂交实验,我们已经确定了这些问题的潜在来源:培养基的制备方案和酵母氮源的来源不仅会影响信号范围,还会影响结果是阳性还是阴性。虽然改变培养基的制备方法可能会优化各个实验的信号差异,但必须详细报告培养基的制备方法,以复制研究并准确比较来自不同实验的结果。