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非标记脱氧核酶构象改变荧光法检测铀酰离子

Spectrofluorimetry determination of UO22+ based on an unlabelled deoxyribozymes conformational switching

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【作者】 曹金秀王永生

【Author】 CAO Jin-xiu;WANG Yong-sheng;College of Public Health,University of South China;

【机构】 南华大学公共卫生学院

【摘要】 在p H 5.5 MES缓冲溶液中,当体系中不存在铀酰离子时,SYBR GreenⅠ(SG)能通过嵌入和小沟结合方式与脱氧核酶作用,导致荧光增强;当铀酰离子存在时,脱氧核酶的r A碱基处断裂,游离出单链,此时SG与单链DNA作用减弱,荧光信号降低,且体系的荧光强度变化值与铀酰离子浓度在1.73×10-9~4.40×10-8mol/L范围内呈良好线性关系,线性回归方程为ΔF=108.99C(×10-8mol/L)+79.22,相关系数r=0.990。根据空白管的标准偏差Sb和标准曲线的斜率k算出LOD为5.2×10-10mol/L。该方法简便、选择性好、灵敏度高。

【Abstract】 In MES buffer solution of p H 5. 5,SG can interact with uranyl specific deoxyribozymes through both intercalation and minor groove binding,resulting in fluorescence enhancement in the absence of UO22 +. The presence of UO22 +causes catalytic cleavage of the DNA substrate strand at the r A position and release of the ss DNA. This leads to the reduction of the interaction of ss DNA with SG,causing a decrease of the fluorescence intensity of the assay system. The decreased fluorescence intensity was proportional to the concentration of uranyl over the range of 1. 73 × 10- 9~ 4. 40 × 10- 8mol / L,and the linear regression equation was ΔF = 108. 99C( × 10- 8mol / L) + 79. 22 with a correlation coefficient of 0. 990.The limit of detection( LOD) was 5. 2 × 10- 10 mol / L. The proposed method was simple,selective and sensitive.

【关键词】 脱氧核酶Sybr GreenⅠ荧光
【Key words】 uranyldeoxyribozymesSybr GreenⅠfluorescence
【基金】 国家自然科学基金资助项目(21177052)
  • 【文献出处】 应用化工 ,Applied Chemical Industry , 编辑部邮箱 ,2015年03期
  • 【分类号】X832;O657.3
  • 【被引频次】3
  • 【下载频次】107
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