项目名称: 中华蜜蜂气味受体基因Or1、Or2的表达、定位与功能研究
项目编号: No.31272513
项目类型: 面上项目
立项/批准年度: 2013
项目学科: 农业科学
项目作者: 姜玉锁
作者单位: 山西农业大学
项目金额: 75万元
中文摘要: 本项目基于我们对中华蜜蜂工蜂气味受体基因AcOr1、AcOr2的前期研究工作(AcOr1为传统气味受体基因,AcOr2为DOr83b直向同源基因),进一步对这两个基因的表达特征进行深入研究,并对其功能进行探索。用qRT-PCR研究雄蜂不同发育阶段两目的基因的表达谱,并与已研究的工蜂的表达特征进行对比。用原位杂交和免疫组化方法对两个目的基因进行定位,分析其mRNA及其蛋白在不同性别中蜂中的表达差异,推测其潜在功能。通过RNAi技术分别抑制AcOr1、AcOr2在活体中的表达,并用qRT-PCR及原位杂交等方法对抑制后的效果进行检测,同时分析两基因间的互作效应。用触角电位仪对干扰组和对照组雌/雄成虫触角对不同气味物质的电生理反应进行测定,分析两气味受体的气味结合特点。本项目可为进一步研究中华蜜蜂特异性气味受体的功能提供分子依据,可望在蜜蜂嗅觉识别蜜粉源机理的研究方面取得新的突破。
中文关键词: 中华蜜蜂;气味受体;基因表达;免疫定位;RNA干扰
英文摘要: Based on our previous research in odorant receptors AcOr1,AcOr2 in Apis cerana cerana, we intend to study the expression characteristics of the two genes in depth and investigate their function in olfactory recognition. Analysing mRNA expresion pattern of drones on different development stages by qRT-PCR and contrasting to that of workers obtained in our previous study. Locating of AcOr1 and AcOr2 in antenna by in siu hybridization and immunohistochemistry to invitagate their expression differences and the potential functions of mRNA and proteins between workers and drones. Silencing the two genes in vivo using RNA interference to detect the inhibition effect and to analyze their interactions. Measuring the electro- phsiological responses of antennae to diverse odors between test and control groups uning electroantennogram (EAG) to analyze the odorant binding characteristics of the two odorant receptors. This project can provide the molecular bases for further study on the specific odorant receptors in A. cerana cerana, and a breakthrough on olfactory recongnition mechanism of honeybees can be expected.
英文关键词: Apis cerana cerana;ordorant receptors;gene expression;immunolocalization;RNAi