Ginkgo Bioworks, Boston, MA, USA.
Methods Mol Biol. 2021;2307:139-145. doi: 10.1007/978-1-0716-1414-3_9.
Homologous recombination is required to specifically target DNA to a desired genomic locus. Non-homologous end joining is the predominant form of recombination in Yarrowia lipolytica. Transformation of this organism with linear DNA therefore results in random integration of the introduced DNA into the genome. In this protocol, hydroxyurea-mediated cell cycle arrest is applied to significantly increase the rate of homologous recombination during transformation and enhance targeted integration.
同源重组是将 DNA 特异性靶向所需基因组位点的必要条件。非同源末端连接是 Yarrowia lipolytica 中主要的重组形式。因此，用线性 DNA 转化该生物体导致引入的 DNA 随机整合到基因组中。在本方案中，应用羟基脲介导的细胞周期阻滞来显著提高转化过程中同源重组的速率，并增强靶向整合。
