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首页> 外文会议>The 6'th International Forum on Post-genome Technologies(6'IFPT)（第六届国际后基因组生命科学技术学术论坛） >CONSTRUCTION, EXPRESSION AND CHARACTERIZATION OF CHIMERA OLFACTORY RECEPTOR
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CONSTRUCTION, EXPRESSION AND CHARACTERIZATION OF CHIMERA OLFACTORY RECEPTOR

机译：希美拉嗅觉受体的构建，表达和鉴定

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摘要

Animals can recognize various chemical compounds by olfactory system. Olfactory cells are equipped with olfactory receptors (ORs), which locate on the cell membranes and bind to chemical compounds specifically. We have been trying to construct chemical sensors by expressing ORs in Saccharomyces cerevisiae. We constructed a chimera olfactory receptor of rat 17 olfactory receptor and mouse mOR226, which recognizes 2, 4-dinitrotoluene (DNT), analogue of trinitrotoluene. Alpha factor receptor, the authentic GPCR, was replaced by the chimera receptor, and GFP or firefly luciferase was introduced as a reporter gene. Introduction and expression of the chimera OR gene was confirmed by PCR and RT-PCR. Then, production of chimera OR was confirmed by western blotting. Finally, we examined the effect of addition of 2, 4-DNT. The transformed yeast cells exhibited dose dependent bioluminescence.
机译：动物可以通过嗅觉系统识别各种化合物。嗅觉细胞配备有嗅觉受体（OR），位于细胞膜上并与化学化合物特异性结合。我们一直在尝试通过在酿酒酵母中表达OR来构建化学传感器。我们构建了大鼠17嗅觉受体和小鼠mOR226的嵌合嗅觉受体，该受体识别2，4-二硝基甲苯（DNT），三硝基甲苯的类似物。真正的GPCRα因子受体被嵌合受体取代，并引入GFP或萤火虫荧光素酶作为报告基因。通过PCR和RT-PCR确认嵌合OR基因的导入和表达。然后，通过western印迹证实嵌合体OR的产生。最后，我们检查了添加2，4-DNT的效果。转化的酵母细胞表现出剂量依赖性的生物发光。

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