IDENTIFICATION OF FALSE POSITIVES IN YEAST TWO-HYBRID DATASETS BY DUAL-REPORTER

点击翻译按钮获取中文摘要

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are attempting to modify the yeast two-hybrid assay such that it can distinguish between bona fide interactions and technical false positives, that is, those positives that do not represent an actual interaction between proteins in the context of the yeast assay. This modification consists of a second reporter protein, in addition to the reconstituted transcriptional activator, that yields a fluorescent signal. The overall idea is to select for positives in a first step based on their ability to activate a reporter gene (HIS3) and allow growth in the absence of histidine, and to screen in a second step for the reconstitution of the signal resulting from the orthogonal reporter.