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嗅觉受体是一种表达在嗅上皮嗅觉受体神经元上的G蛋白偶联受体,可以选择性地与气味分子相结合激活细胞内信号转导通路,使嗅觉受体神经元产生动作电位,在气味识别过程中起重要作用.为了研究嗅觉受体蛋白与气味分子间的相互作用,本研究构建了线虫嗅觉受体ODR-10真核表达载体并将其异源表达于人胚胎肾细胞HEK-293中,通过细胞免疫荧光和Western blot方法对表达效果进行了鉴定.利用细胞内钙离子浓度分析的方法检测表达有嗅觉受体ODR-10的细胞对其天然配体丁二酮的响应,结果证明异源表达的受体依然具有气味识别的功能.将嗅觉受体ODR-10作为传感器的敏感元件固定于石英晶体微天平传感器上,构成质量敏感型气体传感器,该传感器可以检测丁二酮气体,具有很高的灵敏度和特异性,在一定浓度范围内具有线性响应特性.所有结果表明,在异源细胞系统中表达的ODR-10受体蛋白仍具有特异性识别其天然配体的功能,适合作为敏感元件应用于仿生嗅觉受体传感器的构建.
Olfactory receptor is a G protein-coupled receptor expressed on the olfactory epithelium olfactory receptor neurons, which can selectively activate intracellular signal transduction pathway in combination with odorant molecules to make action potential of olfactory receptor neurons , Which plays an important role in odor recognition.In order to study the interaction between olfactory receptor proteins and odorant molecules, we constructed the eukaryotic expression vector of nematode olfactory receptor ODR-10 and expressed heterologous in human embryonic kidney cells The expression of HEK-293 was evaluated by immunofluorescence and Western blot, and the response of cells expressing olfactory receptor ODR-10 to its natural ligand, diacetyl was detected by intracellular calcium ion concentration analysis , The result proved that the heterologously expressed receptor still has the function of odor recognition.The olfactory receptor ODR-10 as the sensor’s sensitive element is fixed on the quartz crystal microbalance sensor to form a mass-sensitive gas sensor which can detect butane di Ketone gas, with a high degree of sensitivity and specificity, has a linear response over a range of concentrations All results indicate that OD expressed in a heterologous cell system R-10 receptor protein still has a specific recognition of its natural ligand function, suitable as a sensing element used in the construction of biomimetic olfactory receptor sensor.