Unnatural amino acid mutagenesis in eukaryotes

真核生物中的非天然氨基酸诱变

• 批准号: 6692253
• 负责人: THOMAS ASHTON CROPP
• 金额: $ 4.16万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-16 至 2005-08-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6692253
• 关键词: active sites; affinity chromatography; aminoacid; aminoacid tRNA ligase; eukaryote; gene mutation; genetic library; mass spectrometry; mutant; polymerase chain reaction; postdoctoral investigator; tissue /cell culture; transfer RNA; yeasts; active sites; affinity chromatography; aminoacid; aminoacid tRNA ligase; eukaryote; gene mutation; genetic library; mass spectrometry; mutant; polymerase chain reaction; postdoctoral investigator; tissue /cell culture; transfer RNA; yeasts

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.

描述（由申请人提供）：拟议的研究旨在扩大在真核细胞中使用的非天然氨基酸诱变的能力。为此，将将正交tRNA/合成酶对引入酵母宿主。然后，将使用特殊设计的遗传选择，将tRNA合成酶的活性位点变体筛选为有趣的非天然氨基酸的新型特异性。一旦确定，新对将使用专门在酵母细胞中的非天然氨基酸位点。稍后，甚至有可能将成功的对将成对的成对移至哺乳动物细胞系。该项目的成功将为膜结合受体的一系列令人兴奋的结构/功能研究和具有新型化学或光谱特性的蛋白质工程打开大门。