Weenink Tim, Ellis Tom
Centre for Synthetic Biology and Innovation, Imperial College London, London, UK.
Methods Mol Biol. 2013;1073:51-60. doi: 10.1007/978-1-62703-625-2_6.
Large numbers of well-described components are essential for advanced synthetic biology and model-guided design of pathways and regulatory networks. Here a method is presented for the creation of libraries of novel control elements. From these libraries, parts with well-defined properties can be selected and used in construction of finely tuned synthetic systems. The example of the PFY1 promoter in S. cerevisiae is used to describe library creation using degenerate synthetic oligos and the circular polymerase extension cloning (CPEC) method. Additionally the workflow of screening the raw library for functional parts is included to provide a full overview of the process of creating and characterizing a component library for synthetic biology.
大量已被充分描述的元件对于先进的合成生物学以及途径和调控网络的模型引导设计至关重要。本文介绍了一种创建新型控制元件文库的方法。从这些文库中，可以选择具有明确特性的元件，并用于构建精细调控的合成系统。以酿酒酵母中的PFY1启动子为例，描述了使用简并合成寡核苷酸和环化聚合酶延伸克隆（CPEC）方法创建文库的过程。此外，还包括对原始文库进行功能元件筛选的工作流程，以全面概述合成生物学中元件文库的创建和表征过程。
