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Investigation of self-assembled protein dimers through an artificial ion channel for DNA sensing

  • Article
  • Analytical Chemistry
  • Published:
Chinese Science Bulletin

Abstract

Artificial nanopores have become promising tools for sensing DNA. Here, we report a new technique for sensing DNA through a conical-shaped nanopore embedded in track-etched polyethylene terephthalate (PET) membrane. Two different streptavidin-conjugated monovalent DNA probes were prepared that can bind to two distinct segments (at either end) of the target DNA. The size of target DNA-linked to the two streptavidin-conjugated monovalent DNA probes is double that of the individual probes. By precisely controlling the tip diameter of the conical nanopore embedded in the PET polymer, events due to the translocation of the streptavidin-conjugated monovalent DNA probes through the nanopore can be filtered and purposely undetected, whereas the current pulses due to the translocation of the target DNA-induced self-assembled complexes can be detected. The two streptavidin-conjugated DNA probes cannot be linked by multi-mismatched DNA. Therefore, multi-mismatched (non-specific) DNA will not induce any current pulse signatures. The current pulse signatures for the self-assembled complex can be used to confirm the presence of the target DNA. The size-dependent detection of self-assembled complexes on the molecular level shows strong promise for the detection of biomolecules without interference from the probes.

摘要

近些年来人工纳米孔材料的设计和开发越来越受到人们的广泛关注,并且有希望应用到DNA的检测中。本文报道了了一个新技术利用锥形纳米孔来测定小片段的DNA。本方法利用目标DNA和与链霉亲和素结合的DNA探针自组装形成蛋白质二聚体,通过电阻脉冲法实现对二聚体的特异性检测。本方法还可以通过改变自组装的对象实现对其他小分子的测定。

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (21190040 and 21275137). The authors would like to thank their supervisor, Charles R. Martin, at the University of Florida. The authors also thank Helmholtzzentrum für Schwerionenforschung GmbH (GSI) for providing track-etched membranes.

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Authors and Affiliations

  1. College of Sciences, Northeastern University, Shenyang, 110819, China

    Siqi Zhang, Ting Sun & Jiahai Wang

  2. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China

    Siqi Zhang, Erkang Wang & Jiahai Wang

Authors
  1. Siqi Zhang
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  2. Ting Sun
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  3. Erkang Wang
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  4. Jiahai Wang
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Correspondence to Jiahai Wang.

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SPECIAL TOPIC: Nanopore Analysis

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Zhang, S., Sun, T., Wang, E. et al. Investigation of self-assembled protein dimers through an artificial ion channel for DNA sensing. Chin. Sci. Bull. 59, 4946–4952 (2014). https://doi.org/10.1007/s11434-014-0626-6

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  • DOI: https://doi.org/10.1007/s11434-014-0626-6

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