Investigation of C-terminal domain of SARS nucleocapsid protein-Duplex DNA interaction using transistors and binding-site models

SENSORS AND ACTUATORS B-CHEMICAL

卷:193

Article

Hsu, You-Ren¹  Kang, Yen-Wen¹  Fang, Jung-Ying¹  Lee, Geng-Yen²  Chyi, Jen-Inn²  Chang, Chung-ke³  Huang, Chih-Cheng¹  Hsu, Chen-Pin¹  Huang, Tai-huang³  Huang, Yu-Fen⁴  Sun, Yuh-Chang⁴  Hsu, Chia-Hsien⁵  Chen, Chih-Chen⁵  Li, Sheng-Shian¹  Yeh, J. Andrew¹  Yao, Da-Jeng¹  Ren, Fan⁶  Wang, Yu-Lin¹ 

[1] Natl Tsing Hua Univ, Inst Nanoengn & Microsyst, Hsinchu 300, Taiwan

[2] Natl Cent Univ, Dept Elect Engn, Tao Yuan 32001, Taiwan

[3] Acad Sinica, Inst Biomed Sci, Taipei 11529, Taiwan

[4] Natl Tsing Hua Univ, Dept Biomed Engn & Environm Sci, Hsinchu 300, Taiwan

[5] Natl Hlth Res Inst, Div Med Engn, Miaoli, Taiwan

[6] Univ Florida, Dept Chem Engn, Gainesville, FL 32611 USA

关键词: SARS;    GaN;    HEMTs;    Sensors;    Dissociation constants;    Binding sites;   

DOI  :  10.1016/j.snb.2013.11.087

来源: Elsevier

【 摘 要 】

AlGaN/GaN high electron mobility transistors (HEMTs) were used to sense the binding between double stranded DNA (dsDNA) and the severe acute respiratory syndrome coronavirus (SARS-CoV) nucleocapsid protein (N protein). The sensing signals were the drain current change of the HEMTs induced by the protein-dsDNA binding. Binding-site models using surface coverage ratios were utilized to analyze the signals from the HEMT-based sensors to extract the dissociation constants and predict the number of binding sites. Two dissociation constants, K-D1 = 0.0955 nM, K-D2 = 51.23 nM, were obtained by fitting the experimental results into the two-binding-site model. The result shows that this technique is more competitive than isotope-labeling electrophoretic mobility shift assay (EMSA). We demonstrated that AlGaN/GaN HEMTs were highly potential in constructing a semiconductor-based-sensor binding assay to extract the dissociation constants of nucleotide-protein interaction. (C) 2013 Elsevier B.V. All rights reserved.

【 授权许可】

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