具有羟乙基团的二氮杂冠醚稀土金属配合物剪切DNA的研究.docVIP

毕业设计（论文） 题目具有羟乙基团的二氮杂冠醚稀 土金属配合物剪切研究 二级学院 化学化工学院 专 业 化学工程与工艺 班 级 学生姓名 学号 才旨导教师 评阅教师 时 间 为了获得能高效稳定地切割DXA的模拟核酸酶，本文根据文献合成了带有羟乙基 支链的1,4, 10, 13-四氧杂-7, 16-二氮杂环十八烷配体(L),将其作为化学核酸酶的 模拟模型，用于质粒超螺旋pUC19DNA的剪切反应；并采用紫外-可见光谱法研究 了稀土金属离子M (Cc31, La3)与配体L形成的稀土金属配合物ML (CcL, LaL)的形 成。通过琼脂糖凝胶电泳法研究Y不同浓度、不同PH、不同反应时间条件K金属配 合物ML (CeL, LaL)对DNA剪切效果的影响，并进一步研究了其作用机理。 实验结果表明，稀土金属离子M (Ce\ La3+)与氮杂冠醚配体形成1:1的金属配 合物ML (CeL, LaL)；琼脂糖凝胶电泳实验表明金属配合物ML (CeL, LaL)能有效地 切割PUC19DNA,且都随着反应时间的增加剪切效果逐渐增强。镧离子配合物(LaL) 在pH值为6. 03左右，浓度为1. 0X10-5 mol/L时对pUC19 DNA的剪切效果最好；铈 离子配合物(CeL)在pH值为7. 04左右，浓度为1. OX 10_5 mol/L时对pUC19 DNA剪 切效果最好。 关键词：二氮杂冠醚；稀土金属离子；镧系金属配合物；剪切DNA Abstract In order to obtain analog nucleases which can cleave DNA stably and efficiently，the complex l，4，10，13- tetraoxa-7,16-diazabicyclo octadecyl ligand (L)with hydroxyethyl was synthesized accoding to the literature，it was used to be the simulation model of metal complexes as chemical nucleases to study the cleavage of supercoiled plasmid pUC19 DNA;Also, the formation of the complex (CeL，LaL ) with rare earth metal ions M (Ce3 +，La3 +) and azamacrocyclic ligand was studied by employing UV-visible spectroscopy.By using agarose gel electrophoresis, we studied the different effects of metal complexes reacted with supercoiled plasmid pUC19 DNA under the different condition of concentrations, PH，reaction time and whether added inhibitors.further more，we explored the reacting maganism of metal complexes cleavage DNA. The experimental results show that rare earth metal ions M (Ce3 \ La3 +) conbined with the aza-crown ether 1:1 to form the metal complexes ML (CeL，LaL) .The experimental results of agarose gel electrophoresis show that metal complexes ML (CeL， LaL) can both accelerated the breakage of pUC19 DNA from supercoiled form (form I) to lined form (form III) effectively，and with the reaction time increasing，the shear effect was enhanced gradually. Lanthanum ion complex (LaL)performed excellent cleavage ability at a pH of about 6.03, a concentration