中文摘要：

【目的】研究中华蜜蜂Apis cerana cerana信息素结合蛋白ASP1与蜜蜂信息素及某些植物挥发物分子的结合功能。【方法】构建中蜂ASP1的原核表达载体，对其进行重组蛋白的诱导表达和分离纯化，并得到具有生化活性的中蜂ASP1重组蛋白，最后以1-NPN作为荧光报告探针，通过荧光竞争结合实验研究中蜂重组ASP1蛋白与蜜蜂信息素及其他气味分子的结合功能。【结果】在22种潜在信息气味物质中，有7种与中蜂ASP1有较强的结合能力，能将1-NPN的相对荧光强度降至50％以下。其中发现蜂王信息素两种成分对-羟基苯甲酸甲酯和香草醇的竞争能力最强，可分别引起1-NPN相对荧光值下降99．31％和95．50％，解离常数KD分别为13．39和98．44μmol／L；而与除蜂王信息素外的其他信息素如幼虫信息素和工蜂信息素等分子均不结合。此外中蜂ASP1对于水杨酸甲酯、苯乙醛、3，4-二甲基苯甲醛4-烯丙基藜芦醚和β-紫罗兰酮等5种植物挥发物质能产生强度不一的结合。【结论】中蜂信息素结合蛋白ASP1对蜂王信息素具有非常强的特异性，同时也能结合某些植物挥发性气味分子，暗示中蜂ASP1是一种以蜂王信息素识别为主要功能、植物挥发物识别为次要功能的多功能信息素结合蛋白。

英文摘要：

[Aim] To study the binding function of Acer-ASP1, a pheromone binding protein （PBP）, in the Chinese honeybee （Apis cerana cerana ） with pheromone and other plant volatiles. [ Methods ] In order to obtain the recombinant protein （Acer-ASP1）, we successfully constructed the cloning and prokaryotic expression vector of Aeer-ASP1, which was expressed in the optimized conditions. After the recombinant protein with biochemical activities was purified, the binding capability of Acer-ASPI with pheromone and other odors was measured using competitive fluorescence assay where 1-NPN was applied as fluorescence probe. [ Results ] Seven of the 22 ligands tested showed stronger binding capability with Aeer-ASP1 and were able to decrease the relative fluorescence intensity of 1-NPN by more than 50%. Among them, methyl 4-hydroxylbenzoate, a queen pheromone, showed the strongest capability to compete with 1-NPN, causing 99.31% reduction in the relative fluorescence intensity, and Ko = 13.39 μmol/L; vanillyl alcohol, another queen pheromone, caused 95.5% decline in the relative fluorescence intensity, and KD = 98.44 μmol/L. Nevertheless, Acer-ASP1 did not bind with other kinds of pheromone at all except queen pheromone. In addition, five plant volatiles, i. e., methyl salicylate, phenylacetaldehyde, 3,4-dimethyl-benzaldehyde, 4-allylveratrole and β-ionone, showed capabilities to bind with ASP1 in various degrees. [ Conclusion ] The results indicate that Acer-ASP1 exhibitsremarkable specificity with queen pheromone, and it can bind with some plant volatiles to some extent. This implies that Acer-ASP1 is probably a protein having complex physiological function, in which recognizing queen pheromone is deemed as its main function while recognizing plant volatiles its secondary function.