Patent ID: 6348315
Filing Date: 2002-02-19
Classification: C07K,C12N

Abstract:
A method for identifying a nucleic acid molecule encoding a peptide or polypeptide that interacts with a target molecule comprising the following steps:(a) transcribing a population of DNA molecules devoid of stop codons in the correct reading frame into the corresponding population of mRNA molecules in the presence of a reducing agent; (b) removing said reducing agent from said population of mRNA molecules; (c) translating said population of mRNA molecules in an in vitro prokaryotic translation system under conditions that allow the formation of polysomes, wherein said translation system either comprises antisense oligonucleotides complementary to the tag-coding sequence of ssrA-RNA or is free of ssrA-RNA; (d) bringing the polysomes so formed into contact with target molecules under conditions that allow the interaction of peptides or polypeptides encoded by said mRNA molecules and displayed by said polysomes with said target molecules; (e) separating polysomes displaying peptides or polypeptides that interact with said target molecules front polysomes displaying no such peptides or polypeptides; and (f) identifying the nucleic acid molecule encoding a polypeptide or peptide displayed in a polysome that interacts with said target molecules.