Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA.
Nat Methods. 2012 Jun;9(6):591-3. doi: 10.1038/nmeth.1971. Epub 2012 Apr 8.
Multiplex automated genome engineering (MAGE) uses short oligonucleotides to scarlessly modify genomes; however, insertions >10 bases are still inefficient but can be improved substantially by selection of highly modified chromosomes. Here we describe 'coselection' MAGE (CoS-MAGE) to optimize biosynthesis of aromatic amino acid derivatives by combinatorially inserting multiple T7 promoters simultaneously into 12 genomic operons. Promoter libraries can be quickly generated to study gain-of-function epistatic interactions in gene networks.
多重自动化基因组工程（MAGE）使用短寡核苷酸无痕修饰基因组；然而，插入长度 >10 个碱基仍然效率低下，但通过选择高度修饰的染色体可以大大改善。本文描述了“共选择”MAGE（CoS-MAGE），通过同时将多个 T7 启动子组合插入 12 个基因组操纵子中来优化芳香族氨基酸衍生物的生物合成。可以快速生成启动子文库，以研究基因网络中功能获得性上位性相互作用。
