Development of a Bacillus subtilis genome vector system that can transmit synthesized genomes

Cloning of small DNA segments has been established using Escherichia coli plasmids. The cloned DNA can be transferred to various cells using transformation. In contrast, cloning of large DNA segments of more than several hundred kilobase pairs has been limited to the Bacillus subtilis genome cloning system. The advantage of giant DNA cloned by B. subtilis is that all kinds of gene editing can be implemented by the high and strict natural transformation ability of the host. However, the following transfer step of giant synthesized and edited genomes to different cell systems requires a special system by avoiding exposure in liquid. The use of a conjugational plasmid pLS20 that was developed for 20 years improves the B. subtilis genome vector establishment process from scratch. The use of the unique B. subtilis genome vector system from synthesis to transmitting genomes is now being manipulated and summarized for the first time.