公开(公告)号：US06448033B1

公开(公告)日：2002-09-10

申请号：US08590729

申请日：1996-01-24

申请人： Wieslaw Kudlicki ,  Gisela Kramer ,  Boyd Hardesty

发明人： Wieslaw Kudlicki ,  Gisela Kramer ,  Boyd Hardesty

IPC分类号： C12P2106

CPC分类号： C07K1/1077 ,  C12N9/10 ,  C12P21/02 ,  G01N33/533

摘要： The present invention provides a method of synthesizing a fluorescently labeled protein in a cell free protein synthesis system, comprising the steps of: (a) incubating a sample of ribosomes obtained from a cell-free extract with plasmid DNA containing a coding sequence for a protein of interest, said sample incubated in a coupled transcription/translation medium together with a aminoacyl tRNA having fluorescent label; (b) partially purifying said fluorescently labeled protein by separating newly synthesized, fluorescently-labeled protein from other fluorescent components within said sample; (c) measuring the amount of protein synthesized; (d) determining fluorescence of newly synthesized protein; and (e) determining the biological activity of the newly synthesized protein.

摘要翻译： 本发明提供了一种在无细胞蛋白质合成系统中合成荧光标记的蛋白质的方法，包括以下步骤：（a）将从无细胞提取物获得的核糖体样品与含有蛋白质编码序列的质粒DNA 所述样品与偶联的转录/翻译培养基一起孵育，同时具有荧光标记的氨酰基tRNA; （b）通过将新合成的荧光标记的蛋白质与所述样品中的其他荧光成分分离，部分纯化所述荧光标记的蛋白质; （c）测量合成的蛋白质的量; （d）确定新合成的蛋白质的荧光; 和（e）确定新合成的蛋白质的生物活性。