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Nearly two decades ago, I started some genetic as well as genomic study using a eubacterium, Bacillus subtilis 168. Her ability drew my great attention, to take up DNA given outside and integrate it into her genome via homologous recombination. This research style later lead to development of a novel cloning vector. Most DNA cloning vectors can not handle a large number of genes, equivalent to giant DNA, at one time. Several works merged to provide the novel cloning system based on the B. subtilis genome as a cloning vehicle. The Bacillus GenoMe (BGM) vector derived from the 4.2-Mb genome of B. subtilsi168 was demonstrated to accommodate fairly large DNAs and is highlighted by the successful stable cloning of a whole
3.5-Mb genome of the nonpathogenic, unicellular photosynthetic bacterium Synechocystis (1) and any sequence-known DNA (2-3). Attempts to build a man-made synthetic genome will also be introduced.
(1) Itaya, M., Tsuge, K. Koizumi, M., and Fujita, K. Combining two genomes in one Cell: Stable cloning of the Synechosystis PCC6803 genome in the Bacillus subtilis 168 genome. Proc. Natl. Acad. Sci., USA., 102, 15971-15976 (2005)
(2) Tsuge, K.、Matsui, K., and Itaya, M. One step assembly of multiple DNA fragments with designed order and orientation in Bacillus subtilis plasmid. Nucleic Acids Res. 31: No. 21, e-133 (2003).
(3) Itaya, M., Fujita, K., Kuroki, A., and Tsuge, K. Bottom-up genome assembly using the Bacillus subtilis genome vector. Nature Methods 5, 41-43 (2008).