Mutagenesis protocols in Saccharomyces cerevisiae by in vivo overlap extension.
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.
Alcalde
Miguel
M
Instituto de Catálisis y Petroleoquímica (CSIC), Madrid, Spain. malcalde@icp.csic.es
D003470:Culture Media; D019020:Directed Molecular Evolution; D016296:Mutagenesis; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D003470:Culture Media