Institute for Advanced Biosciences, Keio University, Nipponkoku, Daihoji, Tsuruoka-shi, Yamagata, Japan.
Department of Bioscience, Tokyo University of Agriculture, Sakuragaoka, Setagaya-ku, Tokyo, Japan.
J Biochem. 2019 Sep 1;166(3):231-236. doi: 10.1093/jb/mvz031.
Bacillus subtilis 168 has been explored as a platform for the synthesis and transmission of large DNA. Two inherent DNA incorporation systems, natural transformation and pLS20-based conjugation transfer, enable rapid handling of target DNA. Both systems are affected by the Bsu restriction-modification system that recognizes and cleaves unmethylated XhoI sites, limiting the choice of target DNA. We constructed B. subtilis 168 with stable mutation for restriction-deficient and modification-proficient (r-m+). It was demonstrated that the r-m+ strains can incorporate and transfer synthesized DNA with multiple XhoI sites. These should be of value as hub strains to integrate and disseminate giant DNA between B. subtilis 168 derivatives.
枯草芽孢杆菌 168 已被探索用于大型 DNA 的合成和传递。两种固有的 DNA 整合系统，自然转化和基于 pLS20 的共轭转移，使目标 DNA 的快速处理成为可能。这两个系统都受到 Bsu 限制修饰系统的影响，该系统识别并切割未甲基化的 XhoI 位点，限制了目标 DNA 的选择。我们构建了枯草芽孢杆菌 168 稳定突变的限制缺陷和修饰高效（r-m+）菌株。结果表明，r-m+菌株可以整合和转移带有多个 XhoI 位点的合成 DNA。这些菌株应该作为中心菌株，在枯草芽孢杆菌 168 衍生物之间整合和传播巨型 DNA。
