Targeted Isolation of a Designated Region of the <i>Bacillus subtilis</i> Genome by Recombinational Transfer

DOI Web Site 被引用文献4件
  • Satoshi Tomita
    Division of Bioscience and Biotechnology, Department of Ecological Engineering, Toyohashi University of Technology, Toyohashi, Aichi 441-8580
  • Kenji Tsuge
    Mitsubishi Kagaku Institute of Life Sciences, Machida-shi, Tokyo 194-8511, Japan
  • Yo Kikuchi
    Division of Bioscience and Biotechnology, Department of Ecological Engineering, Toyohashi University of Technology, Toyohashi, Aichi 441-8580
  • Mitsuhiro Itaya
    Mitsubishi Kagaku Institute of Life Sciences, Machida-shi, Tokyo 194-8511, Japan

抄録

<jats:title>ABSTRACT</jats:title> <jats:p> A method for positional cloning of the <jats:italic>Bacillus subtilis</jats:italic> 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 <jats:italic>ppsABCDE</jats:italic> 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 <jats:italic>ppsABCDE</jats:italic> was constructed using a <jats:italic>B. subtilis</jats:italic> 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 <jats:italic>ppsABCDE</jats:italic> deletion mutant. </jats:p>

収録刊行物

被引用文献 (4)*注記

もっと見る

関連プロジェクト

もっと見る

キーワード

詳細情報 詳細情報について

問題の指摘

ページトップへ