DNA synthesis by fragment assembly using extra-cellular DNA delivered by artificial controlled horizontal transfer
DNA synthesis by fragment assembly using extra-cellular DNA delivered by artificial controlled horizontal transfer
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DOI：
10.1093/jb/mvx085
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发表时间：
2018-04-01
影响因子：
2.7
通讯作者：
Itaya，Mitsuhiro
中科院分区：
文献类型：
--
作者：
Kaneko，Shinya;Fukushima，Hiromi;Itaya，Mitsuhiro
使用细胞外DNA在枯草芽孢杆菌细胞中合成DNA已成为可能。通常，实验室中的分子克隆技术需要在试管中将纯化的DNA引入宿主细胞。我们开发了一种使用稳定的胞外质粒 DNA 进行自然转化的细胞裂解技术，其中胞外质粒 DNA 从裂解的大肠杆菌中释放出来。然后使用稳定的细胞外 DNA 通过片段组装进行 DNA 合成，无需生化纯化。 B 中小鼠线粒体基因组的 DNA 合成。使用 fourE 说明枯草杆菌基因组。带有携带连续 DNA 片段的质粒 DNA 的大肠杆菌菌株。在自然环境中，未纯化的细胞外 DNA 有助于水平基因转移 (HGT) 过程中的基因传递。本研究中引入的技术模仿了 HGT，应该具有广泛的应用。
DNA synthesis in theBacillus subtiliscells has become possible using extra-cellular DNA. Generally, purified DNAs in a test tube have been required to introduce into the host cells for molecular cloning technology in the laboratory. We have developed a cell lysis technique for natural transformation using stable extra-cellular plasmid DNAs, in which the extra-cellular plasmid DNAs are released from lysedEscherichia colicells. DNA synthesis then proceeds by fragment assembly using the stable extracellular DNAs, without biochemical purification. DNA synthesis of the mouse mitochondrial genome inB. subtilisgenome was illustrated using fourE. colistrains with plasmid DNAs carrying contiguous DNA fragments. In the natural environment, unpurified extra-cellular DNAs contribute to the gene delivery during horizontal gene transfer (HGT). The technology introduced in the present study mimics HGT and should have a wide range of applications.