Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Purification of Festriction Endonuclease,SdiI, from Streptomyces diastatochromogenes

Streptomyces diastatochromogens로부터 제한효소 SdiI의 분리정제

  • Bae, Mu (Department of Biological Science. College of Natural Scienc. Ewha Woman's University) ;
  • Song, Eun-Suk (Department of Biological Science. College of Natural Scienc. Ewha Woman's University)
  • 배무 (이화여자대학교 자연과학대학 생물과학과, 서울대학교 분자미생물학연구센터) ;
  • 송은숙 (이화여자대학교 자연과학대학 생물과학과, 서울대학교 분자미생물학연구센터)
  • Published : 1994.01.01
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Abstract

About thirty bacterial strains of actinomycete isolated from the soil were examined for the presence of restriction endonuclease activity. Streptomyces diastatochromogenes, which was identified previously, was found to contain restriction endonuclease activity. The purification of this enzyme, SdiI, was carried out via streptomycin sulfate precipitation and ammonium sulfate fractionation followed by hydroxylapatite column chromatography. Sephacryl S-200 HR column chromatography and second hydroxylapatite column chromatography. SDS-polyacrylamide gel electrophoresis of the active protein (purified from various column chromatography) resulted in 35,000 Da protein.

토양에서 분리한 방선균에서 제한효소 활성을 조사하여 그 가운데 Streptomyces diastatochromogenes로부터 제한효소 활성을 확인하였다. 이 균주의 제한효소 SdiI을 균체 15g으로부터 streptomycin sulfate침전, ammonium sulfate 침전, hydroxylapatite colume chromatography, Sephacryl S-200 HR column chromatography, hydroxylapatite column chromatography의 단계를 거쳐 분리하였고, SDS-polyacrylamide gel-electrophoresis에 의하면 소단위체 분자량은 약 35,000Da으로 추정되었다.

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