Experimental basis for a stable plasmid, pLS30, to shuttle between Bacillus subtilis species by conjugational transfer

Nagayoshi Sakaya, Shinya Kaneko, Satoko Matsunaga, Mitsuhiro Itaya

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The use of Bacillus subtilis 168 as the initial host for molecular cloning and subsequent delivery of the engineered DNA to other Bacillus hosts appears attractive, and would lead to an efficient DNA manipulation system. However, methods of delivery to other Bacillus species are limited due to their inability to develop natural competence. An alternative, unexplored conjugational transfer method drew our attention and a B. subtilis native plasmid, pLS30, isolated from B. subtilis (natto) strain IAM1168 was characterized for this aim. The nucleotide sequence (6,610 bp) contained the mob gene and its recognition sequence, oriT, that features pLS30 as a mobile plasmid between Bacillus species on conjugational transfer. Plasmid pLS3001, a chimera with a pBR322-based plasmid prepared in Escherichia coli to confer an antibiotic resistance marker, showed apparent mobilizing activity in the pLS20-mediated conjugational transfer system recently established. The rep gene and associated palT1-like sequence common to all other pLS plasmids previously sequenced indicated that pLS30 is a typical rolling circle replicating (RCR) type plasmid. Due to the significant stability of pLS30 in IAM1168, application of a mobile plasmid would allow quick propagation to Bacillus species.

Original languageEnglish
Pages (from-to)557-561
Number of pages5
JournalJournal of biochemistry
Volume139
Issue number3
DOIs
Publication statusPublished - 2006 Mar 1

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Keywords

  • Bacillus natto
  • Conjugational transfer
  • Plasmid
  • Rolling circle replication
  • Transformation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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