Alcalde Miguel
Instituto de Catálisis y Petroleoquímica (CSIC), Madrid, Spain.
Methods Mol Biol. 2010;634:3-14. doi: 10.1007/978-1-60761-652-8_1.
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.
高重组频率及其易于操作的特性，使酿酒酵母成为研究同源重组的独特真核模式生物。实际上，酿酒酵母中完善的重组机制有助于构建用于定向进化实验的突变文库。在这种情况下，体内重叠延伸（IVOE）是一种特别有吸引力的方案，它利用真核生物装置进行组合饱和诱变、位点定向重组或位点定向诱变，避免了标准体外方案中常见的连接步骤和额外的PCR反应。
