通过 Golden Gate 组装技术在多拷贝基因座上靶向整合复杂遗传元件。
Targeted Integration of Complex Genetic Elements at Multi-Copy Loci by Golden Gate Assembly.
机构信息
College of Life Sciences, Capital Normal University, Beijing, China.
出版信息
Methods Mol Biol. 2021;2196:143-151. doi: 10.1007/978-1-0716-0868-5_11.
Budding yeast Saccharomyces cerevisiae has become a model eukaryotic microorganism for targeted genomic manipulation due to its efficient homologous recombination. A few genomic loci, including rDNA, Delta, and Ty1, can be utilized to introduce variable copies of genetic elements into the yeast genome. Here we describe a method that combines in vitro Golden Gate Assembly to assemble one or a complex genetic element in an orderly manner and then integrate it into predetermined multi-copy loci through homologous recombination. Different transformants may contain different copy numbers, which allows the selection of desired levels of target gene expression.
出芽酵母酿酒酵母因其高效的同源重组而成为靶向基因组操作的模式真核微生物。一些基因组位点，包括 rDNA、Delta 和 Ty1，可以用于将遗传元件的可变拷贝引入酵母基因组。在这里，我们描述了一种方法，该方法结合体外 Golden Gate 组装，将一个或多个遗传元件有序地组装在一起，然后通过同源重组将其整合到预定的多拷贝位点中。不同的转化体可能含有不同的拷贝数，这允许选择所需的靶基因表达水平。
Zotero 插件