Bxb1 integrase serves as a highly efficient DNA recombinase in rapid metabolite pathway assembly

文献类型： 外文期刊

第一作者： Wang, Xianwei

作者： Wang, Xianwei;Tang, Biao;Mao, Yayi;Lei, Xiaolai;Zhao, Guoping;Ding, Xiaoming;Tang, Biao;Zhao, Guoping;Tang, Biao;Ye, Yu;Zhao, Guoping;Ye, Yu;Zhao, Guoping

作者机构：

关键词： Bxb1 serine integrase;site-specific recombination;DNA assembly;BioBrick

期刊名称：ACTA BIOCHIMICA ET BIOPHYSICA SINICA （ 影响因子：3.848； 五年影响因子：3.67 ）

ISSN： 1672-9145

年卷期： 2017 年 49 卷 1 期

页码：

收录情况： SCI

摘要： Phage-encoded serine integrases are widely used in genetic engineering. They also have the potential to serve as efficient DNA assemblers, demonstrated by the method of site-specific recombination-based tandem assembly (SSRTA) that can combine biological parts into devices, pathways, and systems. Here, four serine integrases, phi BT1, TG1, phi Rv1, and Bxb1, were investigated to ascertain their in vitro DNA assembly activities. Bxb1 integrase displayed the highest efficiency to obtain final products. Thus, we conclude that Bxb1 integrase is an excellent choice for DNA assembly in vitro. Using this enzyme and its recognition sites, BioBrick standards were designed that are compatible with the SSRTA method for module addition. A rapid and efficient procedure was developed for the assembly of a multigene metabolic pathway in one step, directly from non-cutting plasmids containing the gene fragments. This technique is easy and convenient, and would be of interest to the synthetic biology community.

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