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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。
The intricate olfactory system of insects plays a crucial role in the search for hosts, mating, spawning, and evasion by insects. Therefore, the study of insect olfactory perception mechanism will help to reveal the secretions of insects and identify the odor substances in the environment and cause the related behavioral responses. In order to understand more about the interaction mechanism of olotin-related protein in Holotrichia oblita Faldermann in North China, the DUALhunter Starter Kits Yeast Two-Hybrid Screening Kit was used to construct OBP2 decoy odorant binding protein in North China. A homozygous cDNA library of the Megachile giganticolata two - hybrid system. Through the detection of β-galactosidase activity and Blast alignment in GenBank, 6 positive interaction products were identified with OBP2 as bait. We speculate that one of the strong positive interaction clotting zygotes may be the related proteins in the process of olfactory recognition in the genus Coleus.