Molecular Cloning and Partial DNA Sequencing of the Collagenase Gene of <i>Vibrio alginolyticus</i>

J-STAGE
  • FUKUSHIMA Jun
    Department of Bacteriology, Yokohama City University School of Medicine
  • TAKEUCHI Hiroaki
    Department of Bacteriology, Yokohama City University School of Medicine
  • TANAKA Eiichi
    Department of Bacteriology, Yokohama City University School of Medicine
  • HAMAJIMA Kenji
    Department of Bacteriology, Yokohama City University School of Medicine
  • SATO Yayoi
    Department of Bacteriology, Yokohama City University School of Medicine
  • KAWAMOTO Susumu
    Department of Bacteriology, Yokohama City University School of Medicine
  • MORIHARA Kazuyuki
    Institute for Applied Life Science and Department of Enzymology, University of East Asia
  • KEIL Borivoj
    Unit for Chemical Protein, Pasteur Institute
  • OKUDA Kenji
    Department of Bacteriology, Yokohama City University School of Medicine

抄録

DNA fragments Vibrio alginolyticus chemovar iophagus, at least 7kb in length, were ligated to Escherichia coli expression vectors. Three clones of Escherichia coli HB101 (pLCO-1, pLCO-2, pLCO-3) were obtained by the colony immuno-blotting method using anti-collagenase antibody. In Escherichia coli, all these genes produced collagenase antigens which were detected with Western blotting. The amino acid sequence of chemically purified collagenase fragments was also analyzed. An approximately 2.5kb DNA fragment of the pLCO-1 clone was sequenced, and we found that portions of the deduced amino acid sequence were completely identical with the corresponding amino acid sequence of the chemically analyzed fragments. Therefore, it is highly probable that the gene studied in the present experiment is truly a collagenase structural gene.

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