物理化学学报 >> 2006, Vol. 22 >> Issue (08): 917-920.doi: 10.1016/S1872-1508(06)60040-5

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基于SPHD及FRET技术的蛋白质传感器

郭素;廖玮;魏芳;钱民协;赵新生   

  1. 北京分子科学国家实验室, 分子动态与稳态结构国家重点实验室, 北京大学化学学院化学生物学系 北京100871
  • 收稿日期:2006-04-21 修回日期:2006-05-29 发布日期:2006-08-04
  • 通讯作者: 赵新生 E-mail:zhaoxs@pku.edu.cn

Protein Biosensors Based on Scanning Potential Hairpin Denaturation and Fluorescence Resonance Energy Transfer

GUO Su;LIAO Wei;WEI Fang;QIAN Min-Xie;ZHAO Xin-Sheng   

  1. Beijing National Laboratory for Molecular Sciences, State Key Laboratory for Structural Chemistry of Unstable and Stable Species,and Department of Chemical Biology, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R.China
  • Received:2006-04-21 Revised:2006-05-29 Published:2006-08-04
  • Contact: ZHAO Xin-Sheng E-mail:zhaoxs@pku.edu.cn

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摘要：

蛋白质传感器作为一种分析蛋白质与其他分子之间作用力的有效手段已经越来越显示出其优点, 但是蛋白质在基底表面的变性问题对于研究者来说仍然是一个挑战. 作者介绍了一种具有高灵敏度的蛋白质传感器, 即利用DNA引导固定(DDI)方法, 将DNA- 蛋白质复合物通过DNA双链特异性杂交固定在硅基底表面, 从而有效保持了蛋白质的活性, 并且引入荧光共振能量传递(FRET)技术以及电势扫描发卡去杂交(SPHD)技术降低非特异性吸附带来的荧光信号. 这种蛋白质传感器在蛋白质识别以及DNA杂交方面具有显著区分不同体系荧光信号的能力.

关键词: 蛋白质, 传感器, FRET, 电位控制, SPHD, 微阵列

Abstract:

The ability of detecting multiple analytes simultaneously is an important advantage of protein sensors. But the denaturation of proteins on surfaces is still a challenge. In this paper, a protein biosensor providing high selectivity is introduced. This approach is based on DNA-Directed Immobilization (DDI) method, in which protein-DNA conjugates are immobilized on surface through DNA hybridization. Nonspecific binding is reduced by applying fluorescence resonance energy transfer (FRET) and scanning potential hairpin denaturation (SPHD) technologies. Excellent discrimination on different recognition patterns is obtained.

Key words: Proteins, Biosensor, FRET, Potential control, SPHD, Microarray