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

Satoshi Tomita, Kenji Tsuge, Yo Kikuchi, Mitsuhiro Itaya

研究成果: Article査読

15 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)2508-2513
ページ数6
ジャーナルApplied and Environmental Microbiology
70
4
DOI
出版ステータスPublished - 2004 4
外部発表はい

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

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