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 journalArticle

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
Externally publishedYes

Fingerprint

Bacillus subtilis
genome
Genome
DNA
molecular cloning
Organism Cloning
enzyme
genetic vectors
Genetic Vectors
Restriction Mapping
gene
Essential Genes
enzymes
plasmid
antibiotics
plasmids
Plasmids
Anti-Bacterial Agents
Bacteria
mutants

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

Targeted Isolation of a Designated Region of the Bacillus subtilis Genome by Recombinational Transfer. / Tomita, Satoshi; Tsuge, Kenji; Kikuchi, Yo; Itaya, Mitsuhiro.

In: Applied and Environmental Microbiology, Vol. 70, No. 4, 04.2004, p. 2508-2513.

Research output: Contribution to journalArticle

Tomita, Satoshi ; Tsuge, Kenji ; Kikuchi, Yo ; Itaya, Mitsuhiro. / Targeted Isolation of a Designated Region of the Bacillus subtilis Genome by Recombinational Transfer. In: Applied and Environmental Microbiology. 2004 ; Vol. 70, No. 4. pp. 2508-2513.
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