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

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• 作者：You-Ren Hsu ; Yen-Wen Kang ; Jung-Ying Fang ; Geng-Yen Lee ; Jen-Inn Chyi ; Chung-ke Chang ; Chih-Cheng Huang ; Chen-Pin Hsu ; Tai-huang Huang ; Yu-Fen Huang ; Yuh-Chang Sun ; Chia-Hsien Hsu ; Chih-Chen Chen ; Sheng-Shian Li ; J. Andrew Yeh ; Da-Jeng Yao ; Fan Ren ; Yu-Lin Wang
• 关键词：SARS ; GaN ; HEMTs ; Sensors ; Dissociation constants ; Binding sites
• 刊名：Sensors and Actuators B: Chemical
• 出版年：31 March, 2014
• 年：2014
• 卷：193
• 期：Complete
• 页码：334-339
• 全文大小：1339 K

文摘

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.