烟草粉螟性信息素结合蛋白EeluPBP的表达及结合特性  

Expression and binding properties of pheromone binding protein EeluPBP in Ephestia elutella

作　　者：王桂瑶 刘丽[2] 许强[3] 李秋英[4] 朱晋 杨萌萌 王永胜 周汉平[1] 刘英杰[3] 王建伟[1] WANG Guiyao;LIU Li;XU Qiang;LI Qiuying;ZHU Jin;YANG Mengmeng;WANG Yongsheng;ZHOU Hanping;LIU Yingjie;WANG Jianwei(Zhengzhou Tobacco Research Institute of CNTC,Zhengzhou 450001,China;China Tobacco Guangdong Industrial Co.,Ltd.,Guangzhou 510000,China;China Tobacco Jiangsu Industrial Co.,Ltd.,Nanjing 210000,China;Nanping Branch of Fujian Provincial Tobacco Company,Shaowu 354000,Fujian,China)

机构地区：[1]中国烟草总公司郑州烟草研究院,郑州450001 [2]广东中烟工业有限责任公司,广州市510000 [3]江苏中烟工业有限责任公司,南京市210000 [4]福建省烟草公司南平市公司,福建省南平市354000

出　　处：《烟草科技》2024年第5期35-40,共6页Tobacco Science & Technology

基　　金：中国烟草总公司重点研发项目“基于替代磷化铝熏蒸的仓储害虫生态防控技术体系构建及应用”(110202202029);郑州烟草研究院青年人才托举工程项目“烟草粉螟识别性信息素的分子机制研究”(122020CR0240);江苏中烟工业有限责任公司项目“S-烯虫酯控制烟草仓贮害虫综合配套技术研究及应用”(H202223);河南省科技攻关项目“基于知识图谱的烟草病虫害专家系统构建技术研究”(232102210073);广东中烟工业有限责任公司项目“库存偏紧核心产区原料醇化周期研究”(粤烟工科任[2021]-028号)。

摘　　要：为明确烟草粉螟对性信息素的识别机制,在大肠杆菌中表达了烟草粉螟性信息素结合蛋白EeluPBP,通过荧光竞争结合试验分析EeluPBP蛋白与烟草粉螟2种性信息素组分[(顺,反)-9,12-十四碳二烯-1-醇和(顺,反)-9,12-十四碳二烯-1-醇乙酸酯]的结合能力。结果表明:①EeluPBP蛋白在大肠杆菌中成功表达,蛋白质分子量大小约为15 kDa。②EeluPBP蛋白与荧光探针1-NPN的结合解离常数Kd值为0.83μmol/L,表明体外表达的EeluPBP蛋白具有较好的活性。③EeluPBP蛋白与烟草粉螟性信息素组分(顺,反)-9,12-十四碳二烯-1-醇具有较强的结合能力(竞争解离常数Ki值为3.26μmol/L),表明性信息素结合蛋白EeluPBP在烟草粉螟感受性信息素的过程中发挥重要作用。To understand the sex pheromone perception mechanisms in Ephestia elutella,a pheromone binding protein(EeluPBP)from Ephestia elutella was expressed in Escherichia coli in vitro.The binding capacity of EeluPBP protein with the two sex pheromones of E.elutella,(Z,E)-9,12-tetradecadien-1-ol and(Z,E)-9,12-tetradecadien-1-ol acetate,was evaluated by a fluorescence competitive binding assay.The results showed that:1)EeluPBP protein was successfully expressed in E.coli with a molecular weight of approximately 15 kDa.2)The binding dissociation constant Kd value of EeluPBP protein with fluorescent probe 1-NPN was 0.83μmol/L,indicating that the EeluPBP protein expressed and purified in vitro had good binding activities.3)The recombinant EeluPBP protein had a strong binding capacity with(Z,E)-9,12-tetradecadien-1-ol with a Ki value of 3.26μmol/L,indicating that the pheromone binding protein EeluPBP played an important role in the perception of sex pheromone in E.elutella.

关 键 词：烟草粉螟 性信息素结合蛋白 荧光竞争结合试验 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]