Next-generation sequencing coupled with a cell-free display technology for high-throughput production of reliable interactome data.
Next-generation sequencing coupled with a cell-free display technology for high-throughput production of reliable interactome data.
复制标题
新一代测序与无细胞显示技术相结合，可高通量生产可靠的相互作用组数据。
DOI：
10.1038/srep00691
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发表时间：
2012
影响因子：
4.6
通讯作者：
Miyamoto-Sato, Etsuko
中科院分区：
文献类型：
--
作者：
Fujimori, Shigeo;Hirai, Naoya;Ohashi, Hiroyuki;Masuoka, Kazuyo;Nishikimi, Akihiko;Fukui, Yoshinori;Washio, Takanori;Oshikubo, Tomohiro;Yamashita, Tatsuhiro;Miyamoto-Sato, Etsuko
下一代测序技术（NGS）已被应用于各种组学研究，带来了许多生物学和医学发现。然而，来自测序检测的高通量蛋白质-蛋白质相互作用组数据集是稀缺的，因为蛋白质-蛋白质相互作用分析需要许多细胞操作来检查相互作用。高通量数据的低可靠性也是一个问题。在这里，我们描述了一种无细胞显示技术与NGS相结合，可以提高相互作用组数据集的覆盖率和可靠性。完全无细胞的方法提供了高通量和大的检测空间，在不使用克隆的情况下测试相互作用。NGS提供的定量信息减少了假阳性的数量。该方法适用于不仅与诱饵蛋白相互作用，而且与DNA、RNA和化合物相互作用的蛋白质的体外检测。因此，它可能成为探索蛋白质序列和相互作用组网络的大空间的通用方法。
Next-generation sequencing (NGS) has been applied to various kinds of omics studies, resulting in many biological and medical discoveries. However, high-throughput protein-protein interactome datasets derived from detection by sequencing are scarce, because protein-protein interaction analysis requires many cell manipulations to examine the interactions. The low reliability of the high-throughput data is also a problem. Here, we describe a cell-free display technology combined with NGS that can improve both the coverage and reliability of interactome datasets. The completely cell-free method gives a high-throughput and a large detection space, testing the interactions without using clones. The quantitative information provided by NGS reduces the number of false positives. The method is suitable for the in vitro detection of proteins that interact not only with the bait protein, but also with DNA, RNA and chemical compounds. Thus, it could become a universal approach for exploring the large space of protein sequences and interactome networks.