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The Study of Non-enzymatic Cleavage of Oligodeoxyribonucletide by Cerium Complex at Specific Site

Author: ZhouWenJun
Tutor: ShenHeBai
School: Shanghai Normal University
Course: Physical and chemical
Keywords: Artificial endonucleases DNA Oligodeoxynucleotide ODN Triplex DNA DNA sentinel off Surfactant 4 - methyl - 2 6 - bis (N N-di- carboxymethyl ) - methylamino phenol HXTA Binuclear complex Surface -enhanced Raman spectroscopy SERS Fluorescence spectra Fluorescence Spectrum Electrophoresis Silver electrode
CLC: O641.4
Type: Master's thesis
Year: 2003
Downloads: 105
Quote: 1
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Abstract


Artificial enzyme developed under physiological conditions can the efficient hydrolysis cut nucleic acid has a very important significance and application value. Reported at home and abroad have shown that the metal ions and their complexes of nucleic acid phosphodiester bond hydrolysis cut off the role, but their research also just in its infancy, there are many places worthy of further exploration. The paper system to study the oligodeoxynucleotide changes in the state of the electrode surface adsorption oligodeoxynucleotide cerium complexes artificial enzyme the sentinel hydrolysis of the DNA cut role. Many biochemical changes occur in the interface of the charge, so to study the adsorption behavior of the nucleic acids in the charged interface is very necessary. Firstly the length of the DNA fragment and 26 bp to 13 bp length of DNA fragments for the study, and 13-mers DNA in situ electrochemical surface-enhanced Raman spectroscopy and 26-mers DNA in the charged silver electrode adsorbed on the surface state. Determination of the 13-mers DNA and 26-mers DNA on the silver electrode surface-enhanced Raman spectroscopy to study the impact of changes in surface-enhanced Raman spectroscopy of potential changes in the DNA molecule, and speculated that both biological molecules in the silver electrode surface adsorption state with the potential to change the situation; compare the different chain lengths of DNA surface enhanced Raman spectroscopy with potential changes in response rate, and speculated that the reason. We design artificial enzymes consists of two parts, the recognition part, cutting part. Must first want to make to make artificial endonucleases can sentinel cut DNA, identify the target DNA and are firmly bound to the target DNA. We designed the artificial endonuclease recognition section for a DNA fragment complementary to the target DNA nucleotide. In order to study the role of target DNA, we used ion fluorescence probe method as a means of detection, this method has a high sensitivity, fast, simple, practical features. Experimental results show that the synthesis of artificial endonucleases capable of specifically binding to the target DNA. On this basis, the first time we used terbium ion fluorescence probe method to detect the formation of a three-stranded DNA. The well-known three-stranded DNA is the core of the anti-gene and antisense technology, which control gene expression and inhibition, the triple-stranded DNA is currently one of the hot spots. Our experimental results show that the terbium ion fluorescence probe method structure to detect the formation of the three-stranded DNA or not, at the same time, we also study the terbium ions with different types of three-stranded DNA (pyrimidine · purine, pyrimidine, purine · purine · pyrimidine) role the fluorescence spectra of pH level, the presence of metal ions on the three-stranded DNA formed. On the basis of the above studies, we ethylenediamine EDTA covalently connected oligodeoxynucleotide containing 10 bases of the 5 'end, and then in conjunction with the cerium ion, synthetic restriction artificial endonucleases Experimental results show that the restriction within artificial enzymes with identifiable DNA and sentinel hydrolysis to cut off the function of the target DNA, and the cutting mechanism is explained. Furthermore, we also synthesized the 4 - methyl-2, 6 - bis (N, N-di carboxymethyl) - methylamino phenol cerium binuclear complexes, and the cleaving action of the double-stranded DNA in the complexes initial exploration; micelles are ideal cell simulator pieces preliminary study of cerium ions in micellar system of DNA cut.

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