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当前位置: 首页 > 文章 > 酵母双杂交筛选华北大黑鳃金龟触角均一化cDNA文库 昆虫学报 2013,56 (10) 1127-1134
Position: Home > Articles > Screening of the normalized cDNA library from Holotrichia oblita (Coleoptera: Scarabaeida) antennae using yeast two-hybrid system Acta Entomologica Sinica 2013,56 (10) 1127-1134

酵母双杂交筛选华北大黑鳃金龟触角均一化cDNA文库

作  者：
王冰;尹姣;李克斌;曹雅忠
单  位：
中国农业科学院植物保护研究所
关键词：
华北大黑鳃金龟;气味结合蛋白;蛋白互作;酵母双杂交;文库筛选
摘  要：
昆虫错综复杂的嗅觉系统在昆虫寻找寄主、交配、产卵以及逃避行为中起到了至关重要的作用。因此,对昆虫嗅觉感受机理的研究将有助于揭开昆虫感受和识别环境中的气味物质并引起相关行为反应的秘密。为了更多地了解华北大黑鳃金龟Holotrichia oblita Faldermann嗅觉相关蛋白互作机制,本实验利用DUALhunter Starter Kits酵母双杂交筛选试剂盒,构建了华北大黑鳃金龟气味结合蛋白OBP2诱饵载体,筛选了华北大黑鳃金龟触角酵母双杂交系统均一化的cDNA文库。通过β-半乳糖苷酶活性检测以及GenBank中Blast比对分析,以OBP2为诱饵鉴定出6个阳性互作物。我们推测其中的一种较强的阳性互作物凝固酶原可能是华北大黑鳃金龟在嗅觉识别过程中的相关蛋白。
译  名：
Screening of the normalized cDNA library from Holotrichia oblita (Coleoptera: Scarabaeida) antennae using yeast two-hybrid system
作  者：
WANG Bing;YIN Jiao;LI Ke-Bin;CAO Ya-Zhong;State key laboratory for Biology of Plant Disease and Insect Pest,Institute of Plant Protection, Chinese Academy of Agricultural Sciences;
关键词：
Holotrichia oblita;;odorant binding proteins;;protein interaction;;yeast two-hybrid system;;library screening
摘  要：
The sophisticated olfactory system of insects plays a crucial role in their behaviors,such as host seeking,mating,oviposition,as well as escaping behaviors. Therefore,the mystery of arousing a series of insect behavior reactions by perceiving and distinguishing odorants in the environment will be unveiled by investigating olfactory receptive mechanism. In order to clarify the mechanism of interaction between olfactory associated proteins in Holotrichia oblita Faldermann, the HoblOBP2 bait was constructed using DUALhunter Starter Kits. Then,the normalized cDNA library from H. oblita antennae was screened. By detecting β-galactosidase activity and the Blast analysis in GenBank,six interactors were identified using HoblOBP2 bait. We inferred that the proclotting enzyme is the strongest positive interactor,which might play a role in the process of olfactory recognition.

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