Ying Bei-Wen, Akeno Yuya, Yomo Tetsuya
Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University, Osaka, Japan.
Methods Mol Biol. 2013;1073:157-68. doi: 10.1007/978-1-62703-625-2_13.
The construction of stable and functional synthetic circuits in bacteria is necessary in the areas of systems and synthetic biology. The common approach using plasmids to carry foreign genetic circuits offers convenience in genetic construction but is poor in genetic stability (e.g., variation in copy number). Genome recombination provides the stable genetic maintenance of synthetic circuits, but is often labor intensive and time consuming when the genetic circuits are complex and the DNA fragments become larger. The method introduced here is modified from that reported by Wanner's group and is available for integration of complex genetic circuits into the Escherichia coli chromosome.
在系统生物学和合成生物学领域，在细菌中构建稳定且功能正常的合成回路是必要的。使用质粒携带外源遗传回路的常见方法在基因构建方面提供了便利，但遗传稳定性较差（例如拷贝数变化）。基因组重组可实现合成回路的稳定遗传维持，但当遗传回路复杂且DNA片段变大时，通常 labor intensive 且耗时。此处介绍的方法是对Wanner团队报道的方法进行了改进，可用于将复杂遗传回路整合到大肠杆菌染色体中。 （注：原文中“labor intensive”直译为“劳动密集型”，结合语境意译为“操作繁琐、工作量大”更合适）
