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

设计了一种自然转化水平基因转移系统，作为一种无需纯化就能将质粒DNA导入目标细胞的新型克隆工具。本研究确定了供、受体细胞的各培养条件。利用强毒噬菌体诱导供体细胞溶菌具有重要的临床应用价值。结果表明，作为核酸酶抑制剂的金羧酸和作为核酸外切酶促进剂的金羧酸通过适当的混合可以有效地促进DNA的转移。在这种条件下，发现DNA转移扩展到成功的大小增加到100 kb。在试管中对大于100 kb的DNA进行无损处理变得越来越困难。所设计的转移系统称为Culture Mix Method (CMM)，可以简单方便地应用各种质粒dna，这将有助于开发用于合成生物学中基因组合成的动员巨型dna的实验工具。

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.