A New Restriction Endonuclease from Clostridium thermocellum

Clostridium thermocellum으로 부터 새로운 type I I 제한효소 Cth I 의 분리

  • Choi, K.D. (Department of Biological Science and Engineering, Korea Advanced Institute of Science and Technology) ;
  • Kim, Kitae (Department of Biological Science and Engineering, Korea Advanced Institute of Science and Technology) ;
  • Yoo, Ook-Joon (Department of Biological Science and Engineering, Korea Advanced Institute of Science and Technology)
  • Published : 1987.10.01

Abstract

The isolation and characterization of type II restriction endonuclease from Clostridium thermocellum ATCC 27405 were described. This enzyme (Cth I endonuclease) is an isoschizomer of Bcl I endonuclease recognizing 5'-TGATCA-3'. Cth I endonuclease requires MG$^{2+}$ ion for its activity and is maximally active at PH 1.5 to 10.5 in the Presence of 0 to 10mM NaCl. Cth I endonuclease is heat stable and has an optimum temperature of 6$0^{\circ}C$. The activity of Cth I enzyme is sensitive to dam methylation.

새로운 제한효소인 Cth I을 Clostridium thermocellum ATCC 27405로부터 분리하여, 그 효소의 생화학적 특성을 연구하였다. 이 효소는 Bcl I endonuclease의 isoschizomer로서 5'-TGATCA-3'를 인식한다. Cth I endonuclease는 섭씨 60도의 높은 온도에서 잘 작용하며, 10mM까지의 저농도의 NaCl과 MG$^{2+}$ 이온의 존재하에서 pH7.5와 10.5사이에서 최적의 활성을 보여주었다. Cth I endonuclease의 활성은 DNA 기질이 dam methylation 되었을 때 저해를 받았다.

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