采用DATEL方法进行DNA组装。

DNA Assembly with the DATEL Method.

作者信息

Kang Zhen, Ding Wenwen, Jin Peng, Du Guocheng, Chen Jian

机构信息

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.

Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, Jiangsu, China.

出版信息

Methods Mol Biol. 2018;1772:421-428. doi: 10.1007/978-1-4939-7795-6_24.

DOI:10.1007/978-1-4939-7795-6_24

PMID:29754243

Abstract

Simple and reliable DNA assembly methods have become a critical technique in synthetic biology. Here, we present a protocol of the recently developed DATEL (scarless and sequence-independent DNA assembly method using thermostable exonuclease and ligase) method for the construction of genetic circuits and biological pathways from multiple DNA parts in one tube. DATEL is expected to be an applicable choice for both manual and automated high-throughput assembly of DNA fragments, which will greatly facilitate the rapid progress of synthetic biology and metabolic engineering.

摘要

简单可靠的DNA组装方法已成为合成生物学中的一项关键技术。在此，我们展示了一种最近开发的DATEL（使用热稳定核酸外切酶和连接酶的无痕且序列无关的DNA组装方法）方法的方案，用于在一管中从多个DNA片段构建遗传电路和生物途径。DATEL有望成为DNA片段手动和自动高通量组装的适用选择，这将极大地促进合成生物学和代谢工程的快速发展。