Selection of Modified Ribosomes Using Novel Puromycins

项目摘要 这项研究的长期目标涉及修饰肽基转移酶（PTC）中心， 细菌核糖体，使非常不寻常的氨基酸不能识别的野生型， 型核糖体。近年来，我们描述了修饰核糖体的选择和它们的用途 在将D-氨基酸、β-氨基酸、二肽和二肽模拟盒掺入 proteins.在拟议的四年研究期间，工作重点将是选择 能够将磷酸化和糖基化的氨基酸引入特定蛋白质的核糖体。 这种方法的效用将通过分析蛋白质的功能改变来探索， 已经引入了磷酸盐和碳水化合物，并且通过表达磷酸化的DNA， E.杆菌

Project Summary The long term goals of this research involve modification of the peptidyltransferase (PTC) center in bacterial ribosomes to enable the incorporation of very unusual amino acids not recognized by wild- type ribosomes. In recent years, we have described the selection of modified ribosomes and their use in incorporating D-amino acids, beta amino acids, dipeptides and dipeptidomimetric cassettes into proteins. During the four years of proposed research, the focus of efforts will be on the selection of ribosomes capable of introducing phosphorylated and glycosylated amino acids into specific proteins. The utility of this approach will be explored by analyzing the altered function of proteins into which phosphates and carbohydrates have been introduced, and by expressing a phosphorylated DNA binding protein in E. coli.