在采用转录组及蛋白组学方法分别对蝶蛹金小蜂毒腺mRNA特异性表达谱及其毒液蛋白质组成进行分析并鉴定获得毒液富半胱氨酸结构域蛋白（CDPs）基础上，克隆获得编码CDPs的cDNA全序列。分别对其成熟肽编码序列进行原核表达，制备抗体，分析其编码基因的转录与表达的组织特异性及其随寄生蜂发育的动态变化。采用昆虫或酵母表达系统，对CDP编码序列进行表达，研究重组CDPs的功能，及其对寄主菜粉蝶免疫反应及生长发育等重要生理过程的影响。采用免疫共沉淀或酵母双杂交方法、RNA-seq方法、药理学及RNA干扰等系列方法，分别寻找CDPs作用于寄主体内的靶蛋白，分析CDPs处理后寄主相关基因的转录水平变化，明确CDPs作用于寄主体内G蛋白偶联受体类型及其对寄主体内钙离子信号转导途径的调控作用，进而揭示其作用的生理与分子机理。最终，为揭示寄生蜂毒液蛋白组成与其功能，及其调控寄主生理反应的机理提供理论与方法支撑。

In this proposal, we willstudy on the functions and the action modes of cysteine-rich domain-containing proteins (CDPs) isolated from the venom of Pteromalus puparum. Post analyses of the tissue specific expression profile of the mRNA in P. puparum venom gland and the proteins composition of the venom, respectively, the venom CDPs has been identified. For our recent researches, we will first clone the full length cDNAs encoding these CDPs mentioned above. After that, we will recombinantly express the cDNA fragments encoding the mature peptides of these CDPs, respectively, in prokaryotic system, following the preparations of the antiserums against these recombinant proteins. We will investigate the mRNA and protein expression profiles of the CDPs, related to the tissue distribution and the developmental dynamics, by real-time PCR and immunoblotting. Additionally, we will express the sequences encoding the mature peptides of the CDPs utilizing the insect cell or yeast system, and study the functions of the recombinant CDPs and their influences on the importantly physiological processes, including immunity, growth and development of host pest. We will use the methods of Co-IP or yeast two-hybrid, RNA-seq, pharmacological investigation, and RNA interference to search the target proteins of CDPs in the host, to analyze alterations of the transcripts levels of the host related genes, to discover the types of the G protein-coupled receptors in the host, and to illustrate the regulating effects of CDPs on Ca 2+ signaling transduction pathways of the host. Following these researches, we are able to explore the mechanisms at physiological and molecular levels. Finally, these results probably obtained here may provide the new theories and methods to further reveal the compositions and functions of the parasitoid venoms and also the new mechanisms of the venom components regulating the host physiological processes.