CAS Key Laboratory of Quantitative Engineering Biology, Guangdong Provincial Key Laboratory of Synthetic Genomics and Shenzhen Key Laboratory of Synthetic Genomics, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
Methods Mol Biol. 2021;2196:167-180. doi: 10.1007/978-1-0716-0868-5_13.
Budding yeast, as a eukaryotic model organism, has well-defined genetic information and a highly efficient recombination system, making it a good host to produce exogenous chemicals. Since most metabolic pathways require multiple genes to function in coordination, it is usually laborious and time-consuming to construct a working pathway. To facilitate the construction and optimization of multicomponent exogenous pathways in yeast, we recently developed a method called YeastFab Assembly, which includes three steps: (1) make standard and reusable genetic parts, (2) construct transcription units from characterized parts, and (3) assemble a complete pathway. Here we describe a detailed protocol of this method.
budding yeast 作为一种真核模式生物，具有明确的遗传信息和高效的重组系统，是生产外源化学物质的良好宿主。由于大多数代谢途径需要多个基因协调作用，因此构建一个工作途径通常既费力又耗时。为了方便在酵母中构建和优化多组分外源途径，我们最近开发了一种称为 YeastFab 组装的方法，该方法包括三个步骤：（1）制作标准且可重复使用的遗传元件，（2）从已鉴定的元件构建转录单元，以及（3）组装完整的途径。在此，我们详细描述了该方法的步骤。
