• Title/Summary/Keyword: EagBI

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Characterizations of Restriction Endonuclease EagBI from Enterobacter agglomerans CBNU45 (Enterobacter agglomerans CBNU45로부터 분리된 제한효소 EagBI 의 특성)

  • Choe, Yeong-Ju;Kim, Seong-Jae;Hwang, Hye-Yeon;Im, Jeong-Bin;Kim, Yeong-Chang
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.91-95
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    • 1994
  • EagBI is a type II restriction endonuclease from Enterobacter agglomerans strain CBNU45 isolated from soil. EagBI was partially purified by DEAE-cellulose, phosphocellulose P11 and hydroxylapatite column chromatography. EagBI recognizes and cleaves the sequence 5'-CGAT${\downarrow}$CG-3' and generates 2-base 3'-protruding cohesive ends. The optimal reaction conditions of EagBI are 10 mM Tris-HCl (pH 7.8), 6-10 mM $MgCl_2$, at 37 ${\circ}C$. The enzyme is maximally active in the absence of NaCl, able to cleave both $dam^-$ and $dam^+$ DNAs, and sensitive to heat treatment (at 65 ${\circ}C$ for 10 min). Therefore, although EagBI is an isoschizomer of PvuI, it is more useful than PvuI in respect of the NaCl requirement and heat-stability.

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