Unnatural amino acid mutagenesis in eukaryotes

描述（由申请人提供）：拟议的研究旨在扩大非天然氨基酸诱变在真核细胞中的应用。为此，将正交tRNA/合成酶对引入酵母宿主中。然后使用专门设计的遗传选择筛选tRNA合成酶活性位点变体对感兴趣的非天然氨基酸的新特异性。一旦确定，新的对将用于在酵母细胞中特异性地递送非天然氨基酸位点。之后，甚至有可能将成功的配对转移到哺乳动物细胞系中。该项目的成功将为膜结合受体的一系列令人兴奋的结构/功能研究以及具有新化学或光谱特性的蛋白质工程打开大门。

DESCRIPTION (provided by applicant): The proposed research seeks to expand the power of unnatural amino acid mutagenesis to use in eukaryotic cells. In order to do this, an orthogonal tRNA/synthetase pair will be introduced into a yeast host. tRNA synthetase active site variants will then be screened for novel specificities to interesting unnatural amino acids using a specially designed genetic selection. Once identified, the new pair will be used to deliver unnatural amino acids site specifically in yeast cells. Later, it might be possible to even move successful pairs into mammalian cell lines. The success of this project will open the door to a whole array of exciting structure/function studies of membrane bound receptors and the engineering of proteins with novel chemical or spectroscopic properties.