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Molecular Cloning and Partial DNA Sequencing of the Collagenase Gene of Vibrio alginolyticus
Author(s) -
Fukushima Jun,
Takeuchi Hiroaki,
Tanaka Eiichi,
Hamajima Kenji,
Sato Yayoi,
Kawamoto Susumu,
Morihara Kazuyuki,
Keil Borivoj,
Okuda Kenji
Publication year - 1990
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1990.tb01519.x
Subject(s) - vibrio alginolyticus , biology , escherichia coli , microbiology and biotechnology , peptide sequence , collagenase , gene , clone (java method) , nucleic acid sequence , amino acid , dna , molecular cloning , vibrio , biochemistry , genetics , bacteria , enzyme
DNA fragments Vibrio alginolyticus chemovar iophagus , at least 7 kb 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 immunoblotting 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 2.5 kb 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.