Restriction endonuclease in acetic acid bacteria. Part III. Purification, properties and recognition sequence of site-specific restriction endonuclease from Acetobacter liquefaciens AJ 2881.

DOI
  • SASAKI Joji
    Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Shizuoka University
  • YAMADA Yuzo
    Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Shizuoka University

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説明

A type II restriction endonuclease, designated as AliAJI, was purified from cells of Acetobacter liquefaciens AJ 2881 by combined column chromatography on heparin-Sepharose CL-6B, DEAE-Sepharose CL-6B and blue Sepharose CL-6B. The purified enzyme was homogeneous on polyacrylamide gel disc electrophoresis, and the enzyme preparation was free from other nuclease activities, as judged by constancy of lambda DNA-digest electrophoretic patterns after prolonged incubation for 24hr. The enzyme was optimally active at 37°C at pH7.5, required neither sodium chloride nor ammonium sulfate, both of which rather inhibited enzyme activity at high concentration (100 and 75mM, respectively), and cleaved lambda, φX174 RF, SV40, pBR322, M13 mp7 RF and Ad2 DNAs at 18, 1, 2, 1, 1 and 25 or more sites, respectively. The recognition sequence of the enzyme on DNA molecules was determined to be 5'-C-T-G-C-A-G-3', and the enzyme was found to cut between A and G in the sequence, being an isoschizomer of the endonuclease of Providencia stuartii 164 (PstI).

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詳細情報 詳細情報について

  • CRID
    1390001206465229952
  • NII論文ID
    130000027092
  • DOI
    10.1271/bbb1961.48.3027
  • COI
    1:CAS:528:DyaL2MXnsVKisQ%3D%3D
  • ISSN
    18811280
    00021369
  • 本文言語コード
    en
  • データソース種別
    • JaLC
    • Crossref
    • CiNii Articles
  • 抄録ライセンスフラグ
    使用不可

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