中华蜜蜂气味受体AcerOr2的体外表达特性研究

Expression characteristics of olfactory receptor AcerOr2 of Apis cerana cerana in vitro

本研究旨在对中华蜜蜂Apis cerana cerana气味受体基因AcerOr2体外表达特性进行分析验证。本研究利用qRT-PCR和Western blotting检测体外RNA干扰(RNAi)的效果,并通过钙离子成像检测该受体在RNAi前后对气味配体的结合特性。结果显示:体外AcerOr2的最佳干扰浓度为5μg的dsAcerOr2,其能有效抑制目的基因在Sf9细胞中mRNA和蛋白的表达,且AcerOr2本身缺乏对植物气味物质的敏感性,只对其激动剂VUAA1敏感,RNAi后其对VUAA1敏感也有所降低。AcerOr2体外表达特性的验证,可以为进一步研究AcerOr2在嗅觉过程中的功能机制提供基础。

The purpose of this study was to analyze and verify the in vitro expression characteristics of AcerOr2,an olfactory receptor gene of Apis cerana cerana.In this study,qRT-PCR and Western blotting were used to detect the effect of RNA interference(RNAi)in vitro,and the electroantennogram(EAG)was used to detect the binding characteristics of the receptor to odorant ligands before and after RNAi.The results showed that the best interference concentration of AcerOr2 is 5μg ds AcerOr2 in vitro,which can effectively inhibit the mRNA and protein expression of target gene in Sf9 cells.Moreover AcerOr2 itself was not sensitive to plant odorants,but only sensitive to its agonist VUAA1,and its sensitivity to VUAA1 was also reduced after RNAi.The verification of the expression characteristics of AcerOr2 in vitro can provide a basis for further studying the potential functional mechanism of AcerOr2 in the olfactory process.