Designed horizontal transfer of stable giant DNA released from Escherichia coli

Shinya Kaneko, Mitsuhiro Itaya

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

DNA in the environment is a source to mediate horizontal gene transfer (HGT). Present molecular cloning methods are based on this HGT principle. However, DNA in the extracellular environment, particularly with high molecular-weight, is thought to be prone to shearing or digestion by nucleases. Here we discovered that extracellular plasmid DNA released from lysed Escherichia coli remained intact and stable. Furthermore, it was demonstrated that plasmids up to 100 kb in size were taken up by co-present competent Bacillus subtilis cells. The detailed kinetics of the process together with sensitivity to added DNase I indicated that plasmid DNA released from lysed E. coli into the culture medium was stable enough for quantitative efficacy in the transformation of B. subtilis. Our results will be useful for the development of methods to transfer giant DNAs from general host E. coli without their biochemical purification.

Original languageEnglish
Pages (from-to)819-822
Number of pages4
JournalJournal of biochemistry
Volume147
Issue number6
DOIs
Publication statusPublished - 2010 Jun 1

Keywords

  • DNase I sensitivity
  • cell lysis
  • natural genetic transformation
  • phage induction
  • plasmid transfer

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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