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使用在大肠杆菌中表达的嗅觉受体蛋白的压电热敏传感器。

Piezoelectric biosensor using olfactory receptor protein expressed in Escherichia coli.

作者信息

Sung Jong Hwan, Ko Hwi Jin, Park Tai Hyun

机构信息

School of Chemical and Biological Engineering, Seoul National University, Gwanak-Gu Sillim-Dong San 56-1, Seoul 151-744, Republic of Korea.

出版信息

Biosens Bioelectron. 2006 Apr 15;21(10):1981-6. doi: 10.1016/j.bios.2005.10.002. Epub 2005 Nov 16.

Abstract

An olfactory receptor protein of C. elegans, ODR-10, was expressed in Escherichia coli as a fusion protein, with GST and 6x His-tag. The expression of the target protein was analyzed by SDS-PAGE and Western blot, and was confirmed to be expressed at the membrane fraction of the host E. coli. The surface of a quartz crystal microbalance (QCM) was coated with crude membrane extracts, containing the expressed receptor protein, and the interaction between the olfactory receptor and various odorant molecules examined. Compared with other odorants, diacetyl (2,3-butanedione), known as a natural ligand for the ODR-10 receptor, interacted most strongly with the expressed protein. Various concentrations of diacetyl were applied to the expressed ODR-10 receptor, and the response of the QCM showed a linear relationship to the logarithmic value of the odorant concentration. This piezoelectric biosensor system, using olfactory receptor proteins expressed in E. coli, can be used in diagnostics, toxic chemical detection and the quality control of food.

摘要

秀丽隐杆线虫的一种嗅觉受体蛋白ODR-10,作为融合蛋白在大肠杆菌中表达,带有谷胱甘肽S-转移酶(GST)和6x组氨酸标签。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹法(Western blot)分析目标蛋白的表达情况,并证实其在宿主大肠杆菌的膜部分表达。用含有表达的受体蛋白的粗膜提取物包被石英晶体微天平(QCM)的表面,并检测嗅觉受体与各种气味分子之间的相互作用。与其他气味剂相比,作为ODR-10受体天然配体的双乙酰(2,3-丁二酮)与表达的蛋白相互作用最强。将不同浓度的双乙酰应用于表达的ODR-10受体,QCM的响应与气味剂浓度的对数值呈线性关系。这种利用在大肠杆菌中表达的嗅觉受体蛋白的压电生物传感器系统可用于诊断、有毒化学物质检测和食品质量控制。

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