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Helicobacter pylori Strain 51의 보유한 Type II 제한효소 (Hpy51-I)의 특성
조명제,박정욱,전병삼,박정원,변은영,이선경,박예형,송재영,이우곤,백승철,최여정,정선애,최미영,Cho, Myung-Je,Park, Jeong-Uck,Jeon, Beong-Sam,Pack, Jeong-Won,Byun, Eun-Young,Lee, Sun-Kyung,Park, Ye-Hyoung,Song, Jae-Young,Lee, Woo-Kon,Baik, Seun 대한미생물학회 2001 Journal of Bacteriology and Virology Vol.31 No.3
This study describes the purification and characterization of type II restriction endonuclease of Helicobacter pylori in order to understand the DNA restriction and modification of H pylori. H pylori cell extract was subjected to polyethyleneimine treatment, salt precipitation, heparine-sepharose column chromatography, and fast protein liquid chromatogrphy (FPLC) using Resource Q column and Mono Q column to purify the type II restriction endonuclease. Hpy51-I was characterized to recognize the sequneces 5'-GT(G/C)AC-3', yielding 5-base 5' protruding ends. The restriction sequence was identical to that of Tsp 45 I. The enzyme exhibited its maximal activity in the presence of $10{\sim}20\;mM$ NaCl, but was inhibited completely in the presence of more than 80 mM NaCl. The enzyme showed its maximal activity in the presence of $1{\sim}10\;mM\;MgCl_2$. The optimal pH and temperature for enzyme activity was pH 9.0 and $37^{\circ}C$, respectively. $MnCl_2$ could not substitute for $MgCl_2$ in reaction mixture. And addition of ${\beta}$-mercaptoethanol and bovine serum albumin in reaction mixture led to loss of enzyme activity of Hpy51-I. The whole cell extract of H. pylori strain 51 was confirmed to carry the enzyme activity for methylation of Hpy51-I-recognised sequence. Hpy51-I digested genomic DNAs of enteric bacteria to less than I kb while it could not cut the genomic DNAs of H. pylori isolates. In this study, the type II restriction enzyme (Hpy51-I) of H. pylori was identified and characterized its biochemical properties, demonstrating that Hpy51-I might be one of the barriers for preventing the introduction of foreign DNAs into H. pylori.
Helicobacter pylori Strain 51이 보유한 Type Ⅱ
조명제(Myung-Je Cho),박정욱(Jeong-Uck Park),전병삼(Beong-Sam Jeon),박정원(Jeong-Won Pack),변은영(Eun-Young Byun),이선경(Sun-Kyung Lee),박예형(Ye-Hyoung Park),송재영(Jae-Young Song),이우곤(Woo-Kon Lee),백승철(Seung-Chul Baik),최여정(Y 대한미생물학회 2001 Journal of Bacteriology and Virology Vol.31 No.3