Patent ID: 6562594
Filing Date: 2003-05-13
Classification: C12N

Abstract:
A method for producing a set of progeny polypeptides from a template polypeptide, wherein the progeny polypeptides contain a non-stochastic range of single amino acid substitutions represented at each amino acid position, the method comprising:(a) subjecting a codon-containing template polynucleotide to polymerase-based amplification using a 64-fold degenerate oligonucleotide for each codon to be mutagenized, where each of said 64-fold degenerate oligonucleotides is comprised of a first homologous sequence and a degenerate N,N,N triplet sequence, so as to generate a set of progeny polynucleotides; (b) subjecting said set of progeny polynucleotides to clonal amplification such that polypeptides encoded by the progeny polynucleotides are expressed; (c) identifying a first progeny polypeptide having a first amino acid change; (d) identifying a second progeny polypeptide having a second amino acid change; and (e) creating a new progeny polypeptide having both the first amino acid change and the second amino acid change.