高等学校化学学报

高等学校化学学报

• 研究论文 • 上一篇    下一篇

缬氨酸Schiff碱金属铜配合物对质粒DNA的切割作用

王常荣1, 吕健2, 高陆1, 王永梅2, 张金红1   

    1. 南开大学生命科学学院,
    2. 化学学院, 天津 300071
  • 收稿日期:2008-03-18 修回日期:1900-01-01 出版日期:2008-11-10 发布日期:2008-11-10
  • 通讯作者: 张金红

Plasmid DNA Cleavage by Valine-Schiff Base-Cu Complexes

WANG Chang-Rong1, LÜ Jian2, GAO Lu1, WANG Yong-Mei2, ZHANG Jin-Hong1*   

    1. College of Life Science,
    2. College of Chemistry, Nankai University, Tianjin 300071,China
  • Received:2008-03-18 Revised:1900-01-01 Online:2008-11-10 Published:2008-11-10
  • Contact: ZHANG Jin-Hong

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被引次数

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摘要： 缬氨酸Schiff碱金属铜配合物(PBP-L-Val-Cu)是新合成的一类非酶类切割工具, 合成了4种类型样品分别为L-CH3 Cu, D-CH3 Cu, L-Ph Cu和D-Ph Cu. 以质粒DNA(pUC18)为材料,分别对这4种类型化合物进行核酸切割效率的研究, 得出适合的反应体系后, 通过琼脂糖凝胶电泳对反应不同时间后核酸切割产物进行检测, 最终分别得到每种化合物将超螺旋型DNA切割成为开环型DNA和直线型DNA的切割效率, 经比较得出L-CH3 Cu型的切割效率是最快的. 将直线型DNA切割产物用琼脂糖凝胶回收试剂盒进行回收, 得到的直线型切割产物可以在T4连接酶的作用下重新连接起来. 利用酶切法对切口进行检测, 结果表明, 切割作用是具有特异性的. 另外, 该化合物对质粒pNQ216也具有切割活性.

关键词: 缬氨酸Schiff碱金属铜配合物, 质粒DNA, 特异性切割

Abstract: Valine-Schiff base-Cu(PBP-L-Val-Cu), which is a nonenzyme DNA-cleavage complex, contains four different forms: L-CH3, D-CH3, L-Ph and D-Ph. To obtain the most efficient activity, we test the cleavage activity of complexes under different conditions using pUC18 from E.Coli as DNA sample. Plasmid pUC18 was cleaved by the four certain complexes seperately and the products cleaved in variance of time were examined through agarose gel electrophoresis. In this way, we compare the cleavage ability of PBP-L-Val-Cu complexes in cleaving supercoiled DNA into nicked or linear DNA product, and then we find that L-CH3 has the highest activation in cleaving supercoiled plasmid DNA. Futhermore, we purificate the linear DNA product by TIANgel Midi Purification Kit, and this linear DNA extraction could be connected again by T4 DNA ligase. Finally, we incided the linear DNA extraction with two different kinds of restriction enzyme. The results show that this complex can cleave DNA in a site-specific way. Moreover, PBP-L-Val-Cu can also cleave other plasmid DNA, such as pNQ216 from Bacillus subtilis.

Key words: Valine-Schiff base-Cu complex, Plamid DNA, Site-specific cleavage

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