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Study on the Preparation and Application of a Novel Hs-MB Electrochemical Active-inactive Switch Molecular Beacon

Author: WangZuo
Tutor: ChengGuiFang；HePinGang；FangYuZhi
School: East China Normal University
Course: Analytical Chemistry
Keywords: DNA Electrochemistry hemin Molecule Beacon p53 Electro-active/inactive Single-base mismatch
CLC: R346
Type: Master's thesis
Year: 2011
Downloads: 24
Quote: 0
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Abstract


molecules are physically separated, disrupting the hemin dimer and resulting in a doubling of the electrochemical signal. In addition, our design has potential PCR amplification ability for real-world applications with modem simple and portable microelectronics.The dissertation includes three chapters:Chapter 1 PrefaceThe DNA probe based on DNA hybridization technique, with different labeling methods such as chemical luminescent, fluorescent or electrochemical, showing great advantages such as quick detection, easy handling, favorable selectivity. Research on these probes has been one of the most attractive fields in genome study. Taking accounting of the high specificity and selectivity, moleculat beacons have been studied widely. In this chapter, we made a brief introduction of molecular beacons and hemin. It involved the basic principles and relative application, which provided the theoretical principle and selected topic basis for the paper. At last the purpose and innovation of the papar were pointed out.Chapter 2 Study on the electrochemical behavior of hemin and preparation and characterization of a new electrical active-inactive switch molecular beaconIt is well known that hemin is a highly hydrophobic molecule, with poor solubility in neutral and alkaline aqueous buffers. A large number of studies have proved that the physical and chemical properties of aqueous hemin solutions could change in a manner in line with aggregation. In the case of protoporphyrin (the porphyrin moiety in hemin), the tendency to aggregation is quite drastic because of the additional hydrophobic properties of the peripheral vinyl groups. In this chapter, we proved hemin molecular exists clear electrochemical signal differences between the monomer and dimer of hemin in solution, thus, it can be used in our new electrical active-inactive switch molecular beacon. Meantime, all kinds of methods were used to characterize our new electrical active-inactive switch molecular beacon. Indicating that the our molecular possessed excellent ability, which showed potential application value in the field of Biochemistry.Chapter 3 Study on the application of our new Hs-MB electrochemical active-inactive switch molecular beaconP53 gene is an important tumor oncogene. It is reported that 50% human tumors related to p53 gene mutation or loss. So it is very important to analyze and detect p53 gene. In this chapter, a new electrochemical active-inactive switch molecular beacon was synthesized first. This kind of molecular beacon coupled with hemin was electrochemical inactivity, but after hybridized with complementary target sequence, electrochemistry signal was largely enhanced, then applying this molecular beacon in the analysis and detection of p53 target and its SNP gene. The results shown the relationship between the peak currents and the quantity of target. The fitting equation is y=8E-08x+2E-07(X was the concentration of target, M; Y was the peak current, A; R=0.9861), Also, a detection limit of 3nM can be obtained. HS-MB is as effective as conventional molecular beacon in signaling the presence of complementary target and discriminating targets that differ by a single nucleotide. This molecular beacon is fit for quantitative detection and the single-base mismatch recognition of target.

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