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Nucleic Acids Research, 2003, Vol. 31, No. 18 e112
© 2003 Oxford University Press

Conversion of sub-megasized DNA to desired structures using a novel Bacillus subtilis genome vector

Shinya Kaneko, Kenji Tsuge, Takashi Takeuchi and Mitsuhiro Itaya*

Mitsubishi Kagaku Institute of Life Science, 11 Minamiooya, Machida, Tokyo 194-8511, Japan

*To whom correspondence should be addressed. Tel: +81 42 724 6254; Fax: +81 42 724 6316; Email: ita{at}libra.ls.m-kagaku.co.jp

A novel genome vector using the 4215 kb Bacillus subtilis genome provides for precise target cloning and processing of the cloned DNA to the desired structure. Each process highly dependent on homologous recombination in the host B.subtilis is distinguished from the other cloning systems. A 120 kb mouse jumonji (jmj) genomic gene was processed in the genome vector to give a series of truncated sub-megasized DNA. One of these truncated segments containing the first intron was copied in a plasmid by a recombinational transfer method developed for B.subtilis. DNA manipulation previously considered difficult is argued with respect to DNA size and accuracy.


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