Ding Lin, Laor Dana, Weisman Ronit, Forsburg Susan L
Molecular and Computational Biology, University of Southern California, Los Angeles, CA, USA.
Yeast. 2014 Jul;31(7):253-64. doi: 10.1002/yea.3014. Epub 2014 Apr 29.
Genetic analysis of protein function requires a rapid means of inactivating the gene under study. Typically, this exploits temperature-sensitive mutations or promoter shut-off techniques. We report the adaptation to Schizosaccharomyces pombe of the anchor-away technique, originally designed in budding yeast by Laemmli lab. This method relies on a rapamycin-mediated interaction between the FRB- and FKBP12-binding domains to relocalize nuclear proteins of interest to the cytoplasm. We demonstrate a rapid nuclear depletion of abundant proteins as proof of principle.
蛋白质功能的遗传分析需要一种快速使所研究基因失活的方法。通常，这利用温度敏感突变或启动子关闭技术。我们报告了对锚定去除技术的改进，该技术最初由莱姆利实验室在芽殖酵母中设计。此方法依赖于雷帕霉素介导的FRB和FKBP12结合域之间的相互作用，以将感兴趣的核蛋白重新定位到细胞质中。我们证明了大量蛋白质的快速核内耗竭作为原理验证。
