Targeted Isolation of a Designated Region of the Bacillus subtilis Genome by Recombinational Transfer

Satoshi Tomita, Kenji Tsuge, Yo Kikuchi, Mitsuhiro Itaya

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

A method for positional cloning of the Bacillus subtilis genome was developed. The method requires a set of two small DNA fragments that flank the region to be copied. A 38-kb segment that carries genes ppsABCDE encoding five enzymes for antibiotic plipastatin synthesis and another genome locus as large as 100 kb including one essential gene were examined for positional cloning. The positional cloning vector for ppsABCDE was constructed using a B. subtilis low-copy-number plasmid that faithfully copied the precise length of the 38-kb DNA in vivo via the recombinational transfer system of this bacterium. Structure of the copied DNA was confirmed by restriction enzyme analyses. Furthermore, the unaltered structure of the 38-kb DNA was demonstrated by complementation of a ppsABCDE deletion mutant.

Original languageEnglish
Pages (from-to)2508-2513
Number of pages6
JournalApplied and Environmental Microbiology
Volume70
Issue number4
DOIs
Publication statusPublished - 2004 Apr

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

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