One-pot Isolation of Desired Human Genome Fragment by Using Biotinylated pcPNA/S1 Nuclease Combination
One-pot Isolation of Desired Human Genome Fragment by Using Biotinylated pcPNA/S1 Nuclease Combination
复制标题
使用生物素化 pcPNA/S1 核酸酶组合一锅分离所需的人类基因组片段
DOI：
10.1021/acs.biochem.8b00202
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发表时间：
2018
期刊：
影响因子：
2.9
通讯作者：
and Makoto Komiyama
中科院分区：
文献类型：
--
作者：
Arivazhagan Rajendran;Narumi Shigi;Jun Sumaoka;and Makoto Komiyama
人类基因组在预定位点的切割和特定片段的分离对于损伤/修饰位点的分析、蛋白质组学和基因治疗具有极大的意义。然而，用于人类基因组切割和特异性片段分离的方法是有限的。在这里，我们报告了一种新的一锅法，通过使用生物素化的pcPNA/S1核酸酶的组合和所需的片段的链霉亲和素包被的磁珠分离的DNA的位点特异性断裂。概念验证最初被证明用于质粒DNA的剪切和所需片段的分离。然后，我们的方法成功地应用于从细胞来源的人类基因组中分离片段。
Scission of the human genome at predetermined sites and isolation of a particular fragment are of great interest for the analysis of lesion/modification sites, in proteomics, and for gene therapy. However, methods for human genome scission and specific fragment isolation are limited. Here, we report a novel one-pot method for the site-specific scission of DNA by using a biotinylated pcPNA/S1 nuclease combination and isolation of a desired fragment by streptavidin-coated magnetic beads. The proof of concept was initially demonstrated for the clipping of plasmid DNA and isolation of the required fragment. Our method was then successfully applied for the isolation of a fragment from the cell-derived human genome.