Abstract
A high recombination frequency and its ease of manipulation has made Saccharomyces cerevisiae a unique model eukaryotic organism to study homologous recombination. Indeed, the well-developed recombination machinery in S. cerevisiae facilitates the construction of mutant libraries for directed evolution experiments. In this context, in vivo overlap extension (IVOE) is a particularly attractive protocol that takes advantage of the eukaryotic apparatus to carry out combinatorial saturation mutagenesis, site-directed recombination or site-directed mutagenesis, avoiding ligation steps and additional PCR reactions that are common to standard in vitro protocols.
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Acknowledgments
This work was supported by the Spanish Ministry of Science and Innovation (projects CCG08-CSIC/PPQ-3706; PIE 200880I033) and EU project FP7-NMP4-SL-2009-229255.
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Alcalde, M. (2010). Mutagenesis Protocols in Saccharomyces cerevisiae by In Vivo Overlap Extension. In: Braman, J. (eds) In Vitro Mutagenesis Protocols. Methods in Molecular Biology, vol 634. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-652-8_1
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DOI: https://doi.org/10.1007/978-1-60761-652-8_1
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