Purification and Characterization of A Thermotolerable Restriction Endonuclease from Streptomyces violochromogenes D2-5

  • Yun, Mi-Sub (Department of Biological Science, College of natural Science, Ewha Womans University) ;
  • Hwang, Hye-Yeon (Department of Microbiology and Research Center for Molecular Microbiology Seoul National University) ;
  • Bae, Moo (Department of Biological Science, College of natural Science, Ewha Womans University)
  • Published : 1995.10.01

Abstract

A thermotolerable restriction endonuclease. Svil, found in Streptomyces violochromogenes D2-5 was purified. For the purification, streptomycin sulfate and ammonium sulfate precipitation was used. Ph osphocellulose P-ll, DEAE-Cellulose and Sephacryl-S200 HR colum chromatography were also performed. The purified enzyme was found to be homogeneous and the molecular weight of the enzyme estimated by polyacrylamide gel electrophoresis containing 0.1$%$ SDS was about 32, 000 daltons. The recognition sequence and cleavage site of the enzyme were determined to be $5^1$-$TT\downarrow CGAA$-$3^1$ which is the same sequence as that of Asull. Unlike Asull, however, the Svil shows high thermal stability.

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