家蚕hippo信号通路相关基因表达与功能鉴定-expression and functional identification of genes related to hippo signaling pathway in silkworm, bombyx mori.docxVIP

RNAi 后前后翅原基重量变化不大。直观照片显示注射 Yki、Hippo 和 USP 基因 的dsRNA 后家蚕 的前翅原基均显著减小，后翅原基变化不是很明显，其中 Yki 基 因的dsRNA 后效果比 Hippo 和 USP 干扰对翅原基的发育影响更大。 利用病毒介导的超表达方法，调查 Yki 对 家 蚕表型的影响。 转染 pFast-Yki 质粒后，家蚕 Bm12 细胞 Yki 的 mRNA 表达水平上升了 83 倍。转染 Bac-Yki 重组杆状 病毒载体 4 天后，脂肪体中 Yki 转录水平较对照上升了约 3700 倍，翅原基中 Yki 较 对照上升了 283 倍。转染 Bac-Yki 重组杆状病毒载体 4 天后，家蚕前翅原基和后翅原 基并没有发生较大幅度的变化，重量也没有明显变化。这可能与 Yki 必须与其他蛋白 因子共同作用才能调控家蚕翅原基的发育有关。总之，本研究结果证明 Hippo 信号通路调控着家蚕器官的生长发育，其作用受蜕 皮激素信号途径的影响，Yki 基因可以调控家蚕翅原基的发育，但还需要和其他蛋白 因子协同作用。关键词：家蚕；翅原基；Hippo 信号通路； RNAi；过表达Expression and function identification of the genes relat- ed to the Hippo pathway in silkworm, Bombyx moriABSTRACTThe genes of Hippo signaling pathway play important functions in regulation the de- velopment of cell, cell proliferation and cell apoptosis, which also are involved in tumor- igenesis and cancer development. The silkworm, Bombyx mori is a typical economic insect model, with the clear genetic background, which was wildly used as model to analysis the function of the components from agricultural products recently. However, the research about the Hippo singaling pathway in the silkworm has no been reported to now. In the present research, the expression and function identification of the genes related to the Hip- po pathway in silkworm were performed. Our results supply the basis for further eluciatting the mechanism of Hippo pathway in silkworm, and exploitation of silkworm as model for cancer research. In addition, it also constructs the foundation for animal model for screen- ing of the functive componts for cancer therapy.Using Real time-PCR method, the tissues distribution of 16 genes related to hippo pathway in silkworm were performed. Among the 16 genes, about 13 genes were highly expressed in the ovary and wingdisc of Bombyx mori, including Hippo, Sav, Wats, Mats, Kibra, Ex, Mer, Fat, Scribble, Dlg, aPKC, Par6 andYki genes. The Lgl and SD expressed mainly in Prothoracic gland. The Crb gene was highly expressed in the trachea and Hemo- lymph in silkwor