Far rapid synthesis of giant DNA in the Bacillus subtilis genome by a conjugation transfer system

Mitsuhiro Itaya, Mitsuru Sato, Miki Hasegawa, Nobuaki Kono, Masaru Tomita, Shinya Kaneko

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1 Citation (Scopus)

Abstract

Bacillus subtilis offers a platform for giant DNA synthesis, which is mediated by the connection of overlapping DNA segments called domino DNA, in the cloning locus of the host. The domino method was successfully used to produce DNA fragments as large as 3500 kbp. However, domino DNA is limited to <100 kbp because of size restrictions regarding the transformation (TF) of B. subtilis competent cells. A novel conjugal transfer (CT) method was designed to eliminate the TF size limit. The CT method enables rapid and efficient domino reactions in addition to the transfer of giant DNA molecules of up to 875 kbp to another B. subtilis genome within 4 hours. The combined use of the TF and CT should enable significantly rapid giant DNA production.

Original languageEnglish
Article number8792
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - 2018 Dec 1

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Bacilli
Genes
DNA
Cloning
Molecules

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Far rapid synthesis of giant DNA in the Bacillus subtilis genome by a conjugation transfer system. / Itaya, Mitsuhiro; Sato, Mitsuru; Hasegawa, Miki; Kono, Nobuaki; Tomita, Masaru; Kaneko, Shinya.

In: Scientific Reports, Vol. 8, No. 1, 8792, 01.12.2018.

Research output: Contribution to journalArticle

Itaya, Mitsuhiro ; Sato, Mitsuru ; Hasegawa, Miki ; Kono, Nobuaki ; Tomita, Masaru ; Kaneko, Shinya. / Far rapid synthesis of giant DNA in the Bacillus subtilis genome by a conjugation transfer system. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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