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Isolation and characterization of a newly identified type II restriction endonuclease from a local streptomyces sp. in Taiwan

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Abstract

Streptomyces chusanensis ZS-2, isolated from a soil sample in Chusan in Taiwan, was found to produce a new Type II restriction endonuclease. This restriction enzyme was designated as SchI. The purified enzyme was characterized as having a subunit mol wt of 28 kDa, and was apparently free from exonuclease activities. It cleaves the phosphodiester bond between the fourth C and the fifth G on the 5’-CCGCGG-3’ sequence of DNAs, leaving a 2-nucleotide protruding end at its 3’ site. This data suggests that SchI is an isoschizomer of SacII. In addition, based on the comparison between SchI and SacII regarding reaction parameters, it seems that SchI is a better choice of restriction enzyme for genetic analysis and mapping.

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Abbreviations

bp:

base pair(s)

conc:

concentration

EtBr:

ethidium bromide

FPLC:

fast protein liquid chromatography

kDa:

kilodalton(s)

PAGE:

polyacrylamide-gel electrophoresis

PEI:

polyethyleneimine

PMSF:

phenylmethylsulfonyl fluoride

SDS:

sodium dodecyl sulfate

U:

unit(s)

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Authors and Affiliations

  1. Departments of Biochemistry, National Defense Medical Center, Taipei, 100, Taiwan, ROC

    Kuang-Yu Hu, Jia-An Wuu & Ming-Ching Kao

  2. Departments of Microbiology, National Defense Medical Center, Taipei, 100, Taiwan, ROC

    Yu-Tien Liu

  3. Development Center for Biotechnology, Taipei, 100, Taiwan, ROC

    Shou-Hsiung Pai

Authors
  1. Kuang-Yu Hu
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  2. Jia-An Wuu
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  3. Ming-Ching Kao
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  4. Yu-Tien Liu
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  5. Shou-Hsiung Pai
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Hu, KY., Wuu, JA., Kao, MC. et al. Isolation and characterization of a newly identified type II restriction endonuclease from a local streptomyces sp. in Taiwan. Appl Biochem Biotechnol 73, 231–241 (1998). https://doi.org/10.1007/BF02785658

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  • DOI: https://doi.org/10.1007/BF02785658

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