Abstract：
The expansion of the genetic code has enabled the incorporation of noncanonical amino acids (ncAAs) into a defined site of proteins. By introducing such a unique handle into the protein of interest (POI), bioorthogonal reactions can be utilized in live cells to monitor or manipulate the interaction, translocation, function, and modification of the POI. Here, we describe a basic protocol outlining the necessary steps to incorporate a ncAA into a POI in mammalian cells.
摘要：
遗传密码的扩展使得非规范氨基酸（ncAA）能够掺入蛋白质的定义位点。通过将这种独特的手柄引入感兴趣的蛋白质（POI），生物正交反应可用于活细胞中监测或操纵相互作用，易位,函数,和修改的POI。这里,我们描述了一个基本方案，概述了将ncAA掺入哺乳动物细胞POI的必要步骤。
