TTHA氨基酸稀土配合物靶向切割DNA

发布时间：2018-03-21 14:01

  本文选题：TTHA衍生物　切入点：稀土配合物　出处：《内蒙古大学》2017年硕士论文　论文类型：学位论文

【摘要】：本文利用已有的文献和方法首次合成了一系列水溶性很好的三乙四胺六乙酸(TTHA)氨基酸衍生物稀土配合物,稀土元素为镧(La)、铈(Ce)、钕(Nd)、钆(Gd),所用氨基酸为苏氨酸(Thr)、缬氨酸(Val)和谷氨酸(Glu)。该系列稀土配合物通过红外光谱、元素分析和热分析进行实验表征。TTHA氨基酸衍生物稀土配合物与大肠杆菌DNA(pBR322)特异性识别片段相连,再与pBR322进行靶向切割。运用琼脂糖凝胶电泳和原子力显微镜(AFM)的检测手段来探究这一系列配合物对pBR322的作用情况。本文详细研究了 DNA的AFM样的制作过程,此外,通过设计合成靶向探针寡聚核苷酸(ODN)并与TTHA氨基酸衍生物稀土配合物相连,从而形成对pBR322 DNA具有靶向切割作用的稀土配合物,再与pBR322 DNA进行作用。用琼脂糖凝胶电泳从宏观方面讨论了 TTHA氨基酸衍生物稀土配合物对pBR322 DNA的切割作用。通过原子力显微镜在接触模式下对pBR322DNA成像,从微观上进行观测并分析pBR322 DNA构型构象的变化。结果表明在37℃和pH =8.0的生理条件下,合成的一系列稀土配合物能够靶向切割双螺旋DNA。
[Abstract]:In this paper, a series of rare earth complexes of triethylenetetramine hexaacetic acid (TTHAA) amino acid derivatives were synthesized for the first time by using existing literatures and methods. The rare earth elements are lanthanum, cerium, neodymium, neodymium, gadolinium, and amino acids such as threonine, Thyr, valine, valine, and glutamate. The rare earth complexes pass through infrared spectroscopy. Elemental analysis and thermal analysis showed that the rare earth complexes of the amino acid derivatives of TTHA were linked to the specific recognition fragment of Escherichia coli DNA pBR322. Then we used agarose gel electrophoresis and atomic force microscopy (AFM) to investigate the effect of the complexes on pBR322. The preparation of AFM sample of DNA was studied in detail. By designing and synthesizing oligonucleotide oligodeoxynucleotides (ODN) targeting probe and connecting with rare earth complexes of TTHA amino acid derivatives, rare earth complexes with targeted cleavage to pBR322 DNA were formed. The effect of rare earth complexes of TTHA amino acid derivatives on pBR322DNA was discussed macroscopically by agarose gel electrophoresis. PBR322DNA was imaged by atomic force microscope in contact mode. The conformation changes of pBR322 DNA were observed and analyzed microscopically. The results showed that a series of rare earth complexes could target to cut double helix DNAs at 37 鈩，

本文编号：1644162