Patent ID: 7202086
Filing Date: 2007-04-10
Classification: C12N

Abstract:
1. Process for mutagenesis of a target gene which comprises preparing a pool of N oligonucleotides overlapping or not with a sequence substantially complementary to at least one region of the target gene, then reacting said pool of oligonucleotides with said target gene under conditions allowing production of copies of the target gene carrying at least one mutation, wherein the target gene is carried by a double-strand circular plasmid, each oligonucleotide has a sequence complementary to a different region of the target gene and at least one mutation at the centre of the oligonucleotide sequence, the whole of said N oligonucleotides of the pool, with N higher than 5, covering all or part of the sequence of said target gene, and said pool of oligonucleotides is reacted with the target gene in the presence of a polymerase so as to generate a library of mutated genes where each mutation derived from a different oligonucleotide is present on average in less than ⅕ of the genes in the library.