Stable mutants of restriction-deficient/modification-proficient Bacillus subtilis 168: Hub strains for giant DNA engineering

Mitsuhiro Itaya; Mitsuru Sato; Satoru Watanabe; Hirofumi Yoshikawa; Masaru Tomita; Rintaro Sato

doi:10.1093/jb/mvz031

Stable mutants of restriction-deficient/modification-proficient Bacillus subtilis 168: Hub strains for giant DNA engineering

Mitsuhiro Itaya, Mitsuru Sato, Satoru Watanabe, Hirofumi Yoshikawa, Masaru Tomita, Rintaro Sato

Faculty of Environment and Information Studies

Research output: Contribution to journal › Article › peer-review

Abstract

Bacillus subtilis 168 has been explored as a platform for the synthesis and transmission of large DNA. Two inherent DNA incorporation systems, natural transformation and pLS20-based conjugation transfer, enable rapid handling of target DNA. Both systems are affected by the Bsu restriction-modification system that recognizes and cleaves unmethylated XhoI sites, limiting the choice of target DNA. We constructed B. subtilis 168 with stable mutation for restriction-deficient and modification-proficient (r^-m⁺). It was demonstrated that the r^-m⁺ strains can incorporate and transfer synthesized DNA with multiple XhoI sites. These should be of value as hub strains to integrate and disseminate giant DNA between B. subtilis 168 derivatives.

Original language	English
Pages (from-to)	231-236
Number of pages	6
Journal	Journal of biochemistry
Volume	25
Issue number	1
DOIs	https://doi.org/10.1093/jb/mvz031
Publication status	Published - 2020 Jan 1

Keywords

Bacillus subtilis
DNA synthesis
conjugation transfer
genome engineering
natural genetic transformation
restriction-modification

ASJC Scopus subject areas

Biochemistry
Molecular Biology

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10.1093/jb/mvz031

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abstract = "Bacillus subtilis 168 has been explored as a platform for the synthesis and transmission of large DNA. Two inherent DNA incorporation systems, natural transformation and pLS20-based conjugation transfer, enable rapid handling of target DNA. Both systems are affected by the Bsu restriction-modification system that recognizes and cleaves unmethylated XhoI sites, limiting the choice of target DNA. We constructed B. subtilis 168 with stable mutation for restriction-deficient and modification-proficient (r-m+). It was demonstrated that the r-m+ strains can incorporate and transfer synthesized DNA with multiple XhoI sites. These should be of value as hub strains to integrate and disseminate giant DNA between B. subtilis 168 derivatives.",

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author = "Mitsuhiro Itaya and Mitsuru Sato and Satoru Watanabe and Hirofumi Yoshikawa and Masaru Tomita and Rintaro Sato",

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T2 - Hub strains for giant DNA engineering

AU - Itaya, Mitsuhiro

AU - Sato, Mitsuru

AU - Watanabe, Satoru

AU - Yoshikawa, Hirofumi

AU - Tomita, Masaru

AU - Sato, Rintaro

PY - 2020/1/1

Y1 - 2020/1/1

N2 - Bacillus subtilis 168 has been explored as a platform for the synthesis and transmission of large DNA. Two inherent DNA incorporation systems, natural transformation and pLS20-based conjugation transfer, enable rapid handling of target DNA. Both systems are affected by the Bsu restriction-modification system that recognizes and cleaves unmethylated XhoI sites, limiting the choice of target DNA. We constructed B. subtilis 168 with stable mutation for restriction-deficient and modification-proficient (r-m+). It was demonstrated that the r-m+ strains can incorporate and transfer synthesized DNA with multiple XhoI sites. These should be of value as hub strains to integrate and disseminate giant DNA between B. subtilis 168 derivatives.

AB - Bacillus subtilis 168 has been explored as a platform for the synthesis and transmission of large DNA. Two inherent DNA incorporation systems, natural transformation and pLS20-based conjugation transfer, enable rapid handling of target DNA. Both systems are affected by the Bsu restriction-modification system that recognizes and cleaves unmethylated XhoI sites, limiting the choice of target DNA. We constructed B. subtilis 168 with stable mutation for restriction-deficient and modification-proficient (r-m+). It was demonstrated that the r-m+ strains can incorporate and transfer synthesized DNA with multiple XhoI sites. These should be of value as hub strains to integrate and disseminate giant DNA between B. subtilis 168 derivatives.

KW - Bacillus subtilis

KW - DNA synthesis

KW - conjugation transfer

KW - genome engineering

KW - natural genetic transformation

KW - restriction-modification

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Stable mutants of restriction-deficient/modification-proficient Bacillus subtilis 168: Hub strains for giant DNA engineering

Abstract

Keywords

ASJC Scopus subject areas

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