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

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 &...

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Autores principales: Itaya, Mitsuhiro, Sato, Mitsuru, Hasegawa, Miki, Kono, Nobuaki, Tomita, Masaru, Kaneko, Shinya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5993740/
https://www.ncbi.nlm.nih.gov/pubmed/29884789
http://dx.doi.org/10.1038/s41598-018-26987-0
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author Itaya, Mitsuhiro
Sato, Mitsuru
Hasegawa, Miki
Kono, Nobuaki
Tomita, Masaru
Kaneko, Shinya
author_facet Itaya, Mitsuhiro
Sato, Mitsuru
Hasegawa, Miki
Kono, Nobuaki
Tomita, Masaru
Kaneko, Shinya
author_sort Itaya, Mitsuhiro
collection PubMed
description 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.
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spelling pubmed-59937402018-07-05 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 Sci Rep Article 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. Nature Publishing Group UK 2018-06-08 /pmc/articles/PMC5993740/ /pubmed/29884789 http://dx.doi.org/10.1038/s41598-018-26987-0 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
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
title Far rapid synthesis of giant DNA in the Bacillus subtilis genome by a conjugation transfer system
title_full Far rapid synthesis of giant DNA in the Bacillus subtilis genome by a conjugation transfer system
title_fullStr Far rapid synthesis of giant DNA in the Bacillus subtilis genome by a conjugation transfer system
title_full_unstemmed Far rapid synthesis of giant DNA in the Bacillus subtilis genome by a conjugation transfer system
title_short Far rapid synthesis of giant DNA in the Bacillus subtilis genome by a conjugation transfer system
title_sort far rapid synthesis of giant dna in the bacillus subtilis genome by a conjugation transfer system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5993740/
https://www.ncbi.nlm.nih.gov/pubmed/29884789
http://dx.doi.org/10.1038/s41598-018-26987-0
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