| 钟耿军，高旭渊，申建梅，何沐阳，梁康源，张金玟，潘月云，苏湘宁，章玉苹，胡黎明,2024,桔小实蝇BdorOBP2对甲基丁香酚及其结构类似物的结合分析[J].环境昆虫学报,(2):480-488 |
| 桔小实蝇BdorOBP2对甲基丁香酚及其结构类似物的结合分析 |
| Ligand binding properties of odorant binding protein BdorOBP2 to methyl eugenol analogues of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) |
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| DOI： |
| 中文关键词: 桔小实蝇 气味结合蛋白 荧光竞争结合试验 甲基丁香酚类似物 |
| 英文关键词:Bactrocera dorsalis odorant binding protein fluorescence competitive binding assays methyl eugenol analogue |
| 基金项目:广东省普通高校重点领域专项（2021ZDZX4020，2023ZDZX4019）；广东省“仲恺-广东省农业科学院植物保护研究所”联合培养研究生示范基地项目（粤教研函[2021]2号）；2022年广东省“仲恺-广东省科学院南繁种业研究所”联合培养研究生示范基地项目（粤教研函[2022]1号）；广西作物病虫害生物学重点实验室基金（22-035-31-22KF09）;河源市科技计划项目（河科2021002） |
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| 中文摘要: |
| 本文研究了桔小实蝇Bactrocera dorsalis气味结合蛋白BdorOBP2对甲基丁香酚（ME）及其结构类似物的结合特性，以期为基于蛋白结构的新型引诱剂的分子设计提供靶标蛋白。本研究扩增获得气味结合蛋白BdorOBP2基因并进行序列分析。三维结构预测显示，BdorOBP2具有6个α螺旋，同时含有6个保守的半胱氨酸残基。荧光定量结果表明BdorOBP2在雌雄虫触角中的表达水平最高，同时在足、翅等组织中表现出性别差异表达特性，推测BdorOBP2具有识别性信息素和寄主挥发物的双重作用。在对BdorOBP2蛋白表达和纯化的基础上，采用荧光竞争结合试验分析了BdorOBP2与甲基丁香酚及其类似物的结合能力，结果显示BdorOBP2蛋白对丁香酚、异丁香酚、甲基丁香酚、异丁香酚甲醚等4种化合物的结合能力相当。BdorOBP2在感受寄主挥发物及性信息素过程中挥发重要作用，为基于靶标蛋白的新型行为调控剂的分子设计提供了重要参考。 |
| 英文摘要: |
| To provide a basis for the molecular design of novel attractants based on protein structure, this study investigated the binding properties of BdorOBP2 of Bactrocera dorsalis to methyl eugenol (ME) and its structural analogues. The sequence of the BdorOBP2 gene was amplified and analysed. Three-dimensional structure prediction showed that BdorOBP2 has six α-helices and six conserved cysteines. The results of qPCR showed that BdorOBP2 had the highest expression level in antennae of both sexes and showed sex-differential expression characteristics in legs and wings, suggesting that BdorOBP2 plays a dual role in identifying sex pheromone and host volatiles. The recombinant BdorOBP2 was purified, and the binding properties of methyl eugenol and its analogues were studied by fluorescence competitive binding assay. The results showed that BdorOBP2 had a similar binding ability to eugenol, isoeugenol, methyl eugenol, and methyl isoeugenol. BdorOBP2 plays an important role in the volatilisation of host volatiles and pheromones, which provides an important reference for the molecular design of novel behavioral regulators based on target proteins. |
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