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

Research output: Contribution to journalArticle


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 languageEnglish
Pages (from-to)231-236
Number of pages6
JournalJournal of biochemistry
Issue number3
Publication statusPublished - 2019 Sep 1



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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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