Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Characterization of a Restriction Endonuclease, SdiI from Streptomyces diastatochromogenes

Streptomyces diastatochromogenes로부터 분리된 SdiI의 특성에 관한 연구

  • Bae, Moo (Department of Biological Science. College of Natural Scienc. Ewha Woman's University) ;
  • Song, Eun-Sook (Department of Biological Science. College of Natural Scienc. Ewha Woman's University) ;
  • Hwang, Hye-Yeon (Department of Microbiology, College of Natural Sciences, Seoul National University) ;
  • Yim, Jeong-Bin (Department of Microbiology, College of Natural Sciences, Seoul National University)
  • 배무 (이화여자대학교 자연과학대학 생물학과) ;
  • 송은숙 ;
  • 황혜연 (서울대학교 자연과학대학 미생물학과) ;
  • 임정빈 (서울대학교 자연과학대학 미생물학과)
  • Published : 1994.01.01
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Abstract

In catalytic properties of the restriction enonuclease, SdiI, which was purified from Streptomyces diastatochromogenes, this enzyme was active at wide range between pH 7.0 and 12.5, and up to $60^{\circ}C$ and 500 mM of NaCl concentration. It was stable between 20^{\circ}C$ and $60^{\circ}C$, and essentially requires $MgCl_2$ for endonuclease activity. The restriction map of lambda DNA which was obtained by double digestion with various enzymes suggested SdiI to be an isoschizomer of XhoI. From the determination of restriction site based on DNA sequencing method, recognition and cleavage specificity of SdiI was concluded as: 5‘-C${\downarrow}$TCGA G-3' 3'-G AGCT${\uparrow}$C-5'

토양분리 방선균 Streptomyces diastatochromogenes로부터 분리된 제한효소 SdiI은 넓은 범위의 pH(7.0~12.5)와 온도 ($-60^{\circ}C$)에서 활성을 보였으며, 고농도 (-500mM) NaCl이 있어도 작용하였다. 또한, $20~60^{\circ}C$ 온도에서 안정하며, 활성을 갖기 위해서는 $MgCl_2$를 필요로 하였다. Lambda DNA에 대한 지도 작성으로부터 XhoI의 isoschizomer로 추정되었으며, DNa 염기서열 분석 결과, 인식, 절단 서열은 다음과 같이 결정되었다. 5‘-C${\downarrow}$TCGA G-3' 3'-G AGCT${\uparrow}$C-5'

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