SYSTEMS FOR IN VIVO SITE-DIRECTED MUTAGENESIS USING OLIGONUCLEOTIDES

This disclosure provides several methods to generate nucleic acid mutations in vivo, for instance in such a way that no heterologous sequence is retained after the mutagenesis is complete. The methods employ integrative recombinan t oligonucleotides (IROs). Specific examples of the described mutagenesis methods enable site-specific point mutations, deletions, and insertions. Als o provided are methods that enable multiple rounds of mutation and random mutagenesis in a localized region. The described methods are applicable to a ny organism that has a homologous recombination system.