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.

通过自然转化设计的水平基因转移系统已被开发为新的克隆工具，可将质粒DNA引入无纯化而无需纯化。在这项研究中，已经决定了供体和受体细胞的每个培养条件。通过毒性噬菌体对供体细胞的细菌分解诱导对于普遍使用非常重要。此外，据表明，每种甲酸二羧酸作为核酸酶抑制剂和外切核酸酶I作为加速剂，可通过适当地混合DNA转移有效。在这种情况下，发现DNA转移扩展到成功的尺寸增加到100 kb。在试管中，大于100 kb的DNA的无损伤处理越来越困难。所设计的转移系统称为培养混合方法（CMM）可以简单且方便地应用各种质粒DNA，这应该促进实验工具的开发，以动员合成生物学中的基因组合成巨型DNA。

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

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

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