A simple method to provide a shuttling plasmid for delivery to other host ascertained by prolonged stability of extracellular plasmid DNA released from Escherichia coli K12 endA mutant, deficient in major endonuclease

Mitsuhiro Itaya, Yoko Kawata, Mitsuru Sato, Masaru Tomita, Kenji Nakahigashi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Escherichia coli lyses by lambda phage propagation. Circular plasmid DNA was present during E. coli lysis as an extracellular plasmid DNA (excpDNA) that was stable enough to transform coexisting competent Bacillus subtilis cells. Detailed investigations unveiled that excpDNA is transient in both quality and quantity, with stability lasting no more than several hours. A survey using E. coli lambda lysogens with various genetic backgrounds demonstrated that the loss of Endonuclease I (ΔendA::kan) conferred extraordinary stability upon excpDNA for as long as 48 h. Studies on endA mutants suggested that excpDNA remained localized in cell debris, in contrast to E. coli genome DNA, which diffused into medium at an early point in lysis. Lambda lysogens constructed on endA recA mutants are presented for potential pipelines in delivery to other competent proficient microbes.

Original languageEnglish
Pages (from-to)501-504
Number of pages4
JournalJournal of biochemistry
Volume152
Issue number6
DOIs
Publication statusPublished - 2012 Dec 1

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Keywords

  • B. subtilis
  • endA
  • extracellular nucleic acids
  • lambda lysogen
  • transformation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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