Patent ID: 6448033
Filing Date: 2002-09-10
Classification: C07K,C12N,C12P,G01N

Abstract:
A method of synthesizing a fluorescently labeled protein in a prokaryotic, cell-free protein synthesis system, wherein said fluorescent label is on the N-terminus of said protein 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 an 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.