功能化石墨烯的制备及其共振能量转移在生化分析中的应用研究

发布时间：2018-01-11 05:32

  本文关键词：功能化石墨烯的制备及其共振能量转移在生化分析中的应用研究　出处：《西南大学》2012年硕士论文　论文类型：学位论文

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【摘要】：石墨烯自从2004年被发现以来,由于结构、机械、电学及化学特性在场效应晶体管、传感器、清洁能源设备和复合纳米材料等方面有着重要的应用。本文制备了氧化石墨烯,并在此基础上利用石墨烯作为共振能量转移受体,实现了对HIV病毒DNA的分析检测。同时,采用新的方法合成了还原型氧化石墨烯和荧光石墨烯,考察了其物理和化学性质,并进一步探讨了其作为共振能量转移供受体的性质。具体研究内容包括以下三个方面： (1)通过引入氧化石墨烯作为信噪比增强试剂,以人类免疫缺陷病毒1型(HIV-1)相关短DNA序列为例,开发了一种基于长程共振能量转移策略对DNA进行分析检测的简便方法。当体系中不存在靶物ssDNA时,探针ssDNA和Sybr GreenⅠ染料都会吸附到氧化石墨烯表面,此时由于染料和氧化石墨烯之间存在长程共振能量转移过程,所以Sybr GreenⅠ染料荧光被猝灭。当向体系中加入靶物ssDNA,此时靶物ssDNA和探针ssDNA会形成双链结构,由于DNA双链结构和氧化石墨烯较弱的π-π相互作用力,同时Sybr GreenⅠ染料嵌入DNA双链结构的凹槽,所以Sybr GreenⅠ染料的荧光得以恢复,据此高灵敏的检测DNA,检测限能达到0.31nM,并且能够检测碱基错配,该方法相对于其它检测特征DNA序列的方法,具有灵敏、简便、特异性高、抗干扰好的特点。 (2)采用氧化石墨烯固体作为碳源,叶酸作为还原剂,制备了叶酸功能化还原型氧化石墨烯。值得注意的是,叶酸不仅作为还原剂,同时还可作为稳定剂,所制备的还原型氧化石墨烯在水溶液中能够稳定存在4个月以上,进一步从紫外吸收光谱、X射线衍射、红外光谱、拉曼光谱、透射电镜、热稳定性和细胞毒性等方面进行考察,所合成材料表现出较好的还原型氧化石墨烯性质,同时将所制备得到的材料用作共振能量转移受体,和氧化石墨烯进行对比,考察了对荧光素的淬灭,叶酸功能化还原型氧化石墨烯对染料的猝灭效果更好。 (3)以表面活性剂PEG.600作为钝化试剂,开发了一种简单的水热法一步合成荧光石墨烯。由于表面活性剂PEG-600含有大量羟基官能团,所合成的荧光石墨烯能够在水溶液中稳定存在5个月以上,并且其具有较好的抗盐性和抗光漂白性,在pH为2.0-12.0的范围内,荧光强度稳定。
[Abstract]:Graphene has been discovered since 2004, because of structural, mechanical, electrical, and chemical properties of field-effect transistors, sensors. Clean energy equipment and composite nanomaterials have important applications. In this paper graphene oxide was prepared and used as a resonance energy transfer receptor. At the same time, the reduced graphene oxide and fluorescent graphene were synthesized by a new method, and their physical and chemical properties were investigated. The properties of the receptor as a donor of resonant energy transfer are further discussed. The specific contents of the study include the following three aspects: By introducing graphene oxide as a signal-to-noise ratio (SNR) enhancement reagent, the short DNA sequence associated with HIV-1 was used as an example. A simple method for the analysis and detection of DNA based on long range resonance energy transfer strategy is developed, when there is no target ssDNA in the system. The probe ssDNA and Sybr Green I dyes are adsorbed to the surface of graphene oxide, and there is a long range resonance energy transfer process between the dyes and graphene oxide. So the fluorescence of Sybr Green I dye was quenched. When the target DNA was added to the system, the target ssDNA and the probe ssDNA formed double-stranded structure. Because of the weak 蟺-蟺 interaction between DNA double stranded structure and graphene oxide, the Sybr Green 鈪，

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