DNA双受阻胺缺陷的Forster共振能量转移和Laser荧光分析
Forster Resonance Energy Transfer and Laser Fluorescent Analysis of Defects in DNA Double Helix
摘要：
实时激光诱导荧光光谱用于DNA双螺旋缺陷的微分析。该方法基于嵌入剂-供体对（吖啶橙作为供体，溴化乙锭作为受体）中的福斯特共振能量转移（FRET）。过渡金属离子（例如Cu（II）、Cu（I）、Ag（I）、银纳米颗粒（AgNP））、光效应和热效应被用来引起DNA中的双螺旋缺陷。在背景电解质溶液（0.01 M NaNO_3）中实验估计了FRET半径，其值为3.9 ± 0.3 nm，与理论计算值Ro = 3.5 ± 0.3 nm符合良好。以相对单位估计暴露于Cu（II）、Cu（I）、Ag（I）离子、AgNPs冲击以及激光照射（λ = 457 nm）和温度的DNA位点的浓度，其适用于嵌入。FRET方法允许估计双螺旋区域的浓度，具有高质量的稳定性，适用于在DNA受到应力效应后嵌入DNA。它提供了比较1）不同来源的DNA; 2）具有不同损伤程度的DNA; 3）处于各种功能状态的DNA的机会。
英文摘要：
Real time laser induced fluorescence spectroscopy usage for microanalysis of DNA double helix defects is shown. The method is based on Forster resonance energy transfer (FRET) in intercalator-donor pair (acridine orange as a donor and ethidium bromide as an acceptor). Transition metal ions such as Cu(II), Cu(I), Ag(I), silver nanoparticles (AgNPs), photo- and thermo effects were used to cause double helix defects in DNA. FRET radii were experimentally estimated in background electrolyte solution (0.01 M NaNO3) and proved to be 3.9 +- 0.3 nm and the data are in satisfactory agreement with the theoretically calculated value Ro = 3.5 +- 0.3 nm. Concentration of DNA sites, exposed to Cu(II), Cu(I), Ag(I) ions, AgNPs impact as well as laser irradiation (λ = 457 nm) and temperature, which are applicable for intercalation, were estimated in relative units. FRET method allows to estimate the concentration of double helix areas with high quality stability applicable for intercalation in DNA after it was subjected to stress effect. It gives the opportunity to compare DNAs of 1) different origin; 2) with various damage degrees; 3) being in various functional state.
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