A Nobel cloning tool using the designed horizontal gene-transfer system by natural transformation

诺贝尔克隆工具，使用通过自然转化设计的水平基因转移系统

• 批准号: 23710239
• 负责人: KANEKO Shinya
• 金额: $ 2.91万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2013
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23710239/
• 关键词: 応用微生物; 核酸; ゲノム; バイオテクノロジー; 形質転換

A designed horizontal gene-transfer system by natural transformation has been developed as novel cloning tool to introduce plasmid DNA into the objective cells without purification. Each culture condition for donor and recipient cells has been decided in this study. The bacteriolysis induction of donor cells by virulent phage is very important for universal use. Furthermore it was revealed that each aurintricarboxylic acid as nuclease inhibitor and exonuclease I as accelerator is effective for the DNA transfer by mixing it appropriately. In this condition it was found that the DNA transfer was extended to successful size increase up to 100 kb. The damage-free handling of DNA larger than 100 kb in test tubes is increasingly difficult. The designed transfer system named as Culture Mix Method (CMM) can apply various plasmid DNAs simply and conveniently, which should facilitate development of experimental tools to mobilize giant DNAs for genomic syntheses in synthetic biology.

设计了一种水平基因转移系统，通过自然转化将质粒DNA导入目的细胞而无需纯化，是一种新型的克隆工具。本研究确定了供体和受体细胞的各种培养条件。用强毒噬菌体诱导供体细胞的溶菌作用对于普遍应用是非常重要的。此外，揭示了作为核酸酶抑制剂的每种金精三羧酸和作为加速剂的核酸外切酶I通过将其适当混合而对DNA转移有效。在这种条件下，发现DNA转移被扩展到成功的大小增加到100 kb。在试管中无损处理大于100 kb的DNA越来越困难。所设计的质粒转移系统CMM（CultureMixMethod）可以简单方便地应用于各种质粒DNA，为合成生物学中基因组合成的大规模DNA转移提供了实验工具。

项目成果

細胞外核酸による水平伝播システム(CMM)を用いたゲノム合成 A designed horizontal gene-transfer system by natural transformation, named as Culture Mix Method (CMM) for genomic syntheses

通过自然转化设计的水平基因转移系统，称为基因组合成的培养混合法（CMM）

• 发表时间: 2012
• 作者: 金子真也;板谷光泰
• 通讯作者: 板谷光泰

水平伝播を利用したクローニングシステム(CMM)の汎用性 A versatile designed horizontal gene-transfer system by natural transformation, named as Culture MixMethod (CMM) for genomic syntheses

一种通过自然转化设计的多功能水平基因转移系统，称为用于基因组合成的培养混合方法（CMM）

• 发表时间: 2014
• 作者: 金子真也;板谷光泰
• 通讯作者: 板谷光泰

細胞外核酸の水平伝播を介するゲノム合成 Genome synthesis via horizontal transfer of stable extra-cellular DNA

通过稳定的细胞外 DNA 水平转移进行基因组合成

• 发表时间: 2011
• 作者: 板谷光泰;金子真也
• 通讯作者: 金子真也