单晶硅基底薄膜中DNA与不同结构球状蛋白的相互作用

L. Butusov, I. Nagovitsyn, V. V. Kurilkin, Chudinova Gk
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引用次数: 3

摘要

用荧光光谱法研究了小牛胸腺DNA与兔免疫球蛋白(IgG)、高铁血红蛋白(MHB)、人血清白蛋白(HSA)等不同蛋白混合后在单晶硅衬底上的双组分薄膜。在340 ~ 380 nm范围内,用λex=260和280 nm对荧光光谱进行配准。采用旋涂法制备薄膜。沉积在底物上的溶液含有相同数量DNA的10-9 - 10-15 M的小浓度蛋白质。每种使用的蛋白质的荧光强度与浓度的形状依赖性明显不同。随着膜中蛋白质浓度的降低,DNA-HSA膜的综合荧光强度(与10-9 M浓度相比)分别为2.5倍和5倍(10-13和10-15 M HSA), DNA-IgG膜的综合荧光强度为4.6倍和15.9倍(10-11和10-13 M MHB), DNA-MHB膜的综合荧光强度为3.4倍(10-11 M MHB)。以单组分膜作为对照样品进行研究,其性质与dna -蛋白质系统的性质明显不同。dna -蛋白膜对不同结构、不同浓度蛋白质荧光特性的特异性,可作为开发生物传感器系统的基础。
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Interaction of DNA with Globular Proteins of Different Structures in Thin Films on Substrates of Monocrystalline Silicon
The two-component films of mixtures of DNA (from calf thymus) with different proteins: rabbit immunoglobulin (IgG), methemoglobin (MHB) and human serum albumin (HSA) were studied on single-crystal silicon substrates by the method of fluorescence spectroscopy. Registration of fluorescence spectra was performed by λex=260 and 280 nm in the range 340-380 nm. To prepare films the spincoating method was used. Solutions deposited on the substrate contained the small concentrations of proteins 10-9 - 10-15 M at the same quantity of DNA. Shape dependencies of the fluorescence intensity versus concentration differ markedly for each of the used proteins. The decreasing concentration of protein in the film is accompanied by a significant increase of the integrated fluorescence intensity (in comparison with the concentration of 10-9 M) for films of DNA-HSA in 2.5 and 5 times (10-13 and 10-15 M HSA), for films of DNA-IgG in 4.6 and 15.9 times (10-11 and 10-13 M IgG), for films of DNA-MHB in 3.4 times (10-11 M MHB). The single-component films of proteins was studied as control samples whose properties noticeably differ from the properties of DNA-protein systems. The specificity of the fluorescent characteristics of DNA-protein films for proteins of different structure and their different concentrations could be used as the basis for developing biosensor systems.
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