蝶蛹金小蜂毒液免疫抑制小肽的鉴定、结构与功能研究

项目名称： 蝶蛹金小蜂毒液免疫抑制小肽的鉴定、结构与功能研究

项目编号： No.31272098

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 叶恭银

作者单位： 浙江大学

项目金额： 80万元

中文摘要： 采用小肽分离与鉴定、基因克隆与表达、生物活性测定及小肽结构解析等技术，在明确菜粉蝶重要寄生蜂蝶蛹金小蜂毒液中免疫抑制小肽组成的基础上，筛选并克隆免疫抑制小肽的编码基因，探明免疫抑制小肽编码基因转录与表达水平的时空变化；进而采用酵母表达系统对免疫抑制小肽进行表达（或采用化学法对其进行合成），测定免疫抑制小肽生物学功能，并解析其结构，研究结构与功能间关系；最后对免疫抑制小肽与凝集素GNA的融合蛋白进行表达，测定其对寄主与非寄主生理功能的影响。最终，明确该蜂毒液免疫抑制小肽的结构与功能，并探明其在害虫生防中的应用前景。一方面为深入揭示寄生蜂毒液组成与功能，及功能组分调控寄主先天免疫反应的机理提供新思路与新方法；另方面为建立以害虫免疫系统为靶标的新型杀虫剂－免疫抑制剂的筛选与研发平台，及基于抑制免疫反应的新型害虫生防策略研究奠定理论基础与技术支撑，进而为"绿色农业"可持续发展提供害虫控制的新途径。

中文关键词： 寄生蜂；毒液小肽；结构；功能；寄主免疫

英文摘要： For the experiments that will be carried out in this project, series of technological methods will be used, including peptide purification and identification, gene cloning and recombinant expression, biological activity assay, and protein structural analysis. Based on the composition of small immunosuppressive peptides from the venom of a pupal stage endoparasitoid, Pteromalus puparum, of its host Peris rapae, the genes encoding small immunosuppressive peptides will be first screened and cloned. Second, the transcripts and expression levels of different peptide genes in different tissues and developmental stages of the parasitoid will be investigated. Third, the selective small peptides will be expressed in yeast system (or be chemically synthesized) and the biological functions of their recombinant or synthesized products will be assayed. Forth, the structures of the small peptides will be analyzed and the relationship between structure and function will also be studied. Fifth, the small peptides will be recombinantly expressed with a fusion protein, Galanthus nivalis agglutinin (GNA), and the influences of the fusion peptides on the physiologicail aspects of host and no-host insects will be determined. Finally, the structural and functional compositions of small immunosuppressive peptides from parasitoid venom

英文关键词： Parasitoid wasps；Venom peptides；Structures；Functions；Host immunity