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

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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.