Kearsey Stephen E, Gregan Juraj
Department of Zoology, University of Oxford, Oxford, UK.
Methods Mol Biol. 2009;521:483-92. doi: 10.1007/978-1-60327-815-7_27.
Inactivating a specific protein in vivo can yield important information about its function. One strategy previously developed in Saccharomyces cerevisiae by the Varshavsky group involves fusing a degron, derived from mouse dihydrofolate reductase, to the N-terminus of the target protein, which thereby confers temperature-sensitive degradation at the restrictive temperature. We describe here the application of this technique in the fission yeast, Schizosaccharomyces pombe.
在体内使特定蛋白质失活可以产生有关其功能的重要信息。瓦尔沙夫斯基小组先前在酿酒酵母中开发的一种策略是将源自小鼠二氢叶酸还原酶的降解结构域融合到目标蛋白质的N端,从而在限制温度下赋予温度敏感型降解特性。我们在此描述了该技术在裂殖酵母粟酒裂殖酵母中的应用。