Research Center of Biological Information, College of Science, National University of Defense Technology, Changsha, Hunan 410073, China.
Beijing Institute of Pharmaceutical Chemistry, Beijing 102205, China.
Sci Rep. 2017 Mar 16;7:44648. doi: 10.1038/srep44648.
Here, an efficient cloning strategy for large DNA fragments and for simultaneous assembly of multiple DNA fragments assembly is presented. This strategy is named OEPR (based on Overlap Extension PCR and Recombination in vivo). OEPR cloning is a seamless, restriction- and ligation-independent method. The method takes advantage of both homologous recombination enzymes in E. coli and overlap PCR. Using OEPR cloning, a long fragment (1-6 kb) or multiple fragments (2-4 fragments) can be easily constructed and simultaneously assembled into a target vector.
这里提出了一种用于大片段 DNA 克隆和多个 DNA 片段同时组装的有效克隆策略。该策略命名为 OEPR（基于重叠延伸 PCR 和体内重组）。OEPR 克隆是一种无缝、无限制酶切和连接的方法。该方法利用了大肠杆菌中的同源重组酶和重叠 PCR。使用 OEPR 克隆，可以轻松构建长片段（1-6kb）或多个片段（2-4 个片段），并同时将它们组装到目标载体中。
