棉铃虫Fox-uc蛋白的功能及其调控CYP6B6基因的表达

在寄主植物-昆虫-杀虫剂的相互作用网络中，细胞色素P450酶系参与棉铃虫对多种植物次生物质和杀虫剂的解毒代谢过程。在前期研究2-十三烷酮(2-TD)诱导下，参与棉铃虫CYP6B6过量表达的调控蛋白时，发现一个能与CYP6B6启动子结合的新转录因子，即叉头框未分类蛋白(Fox-uc)。在此基础上，本项目将研究棉铃虫Fox-uc的功能，及其在2-十三烷酮胁迫后调控CYP6B6表达的过程。拟用RNA干扰技术检测Fox-uc在棉铃虫生长和解毒过程中的作用，用凝胶阻滞技术测定Fox-uc在CYP6B6启动子上的结合位点，用染色质免疫共沉淀技术确定在2-TD胁迫下Fox-uc是调控CYP6B6表达的抑制因子。本项目将进一步完善CYP6B6响应植物次生物质胁迫的调控机制，这对掌握棉铃虫P450的解毒机制具有重要意义，并为在基因调控水平防治棉铃虫提供新的研究靶点。

In the interactive network of plant-insect-insecticide, the cytochrome P450 enzymes are involved in the detoxification and metabolism responded to various plant secondary substances and insecticide in Helicoverpa armigera. In previously study of the CYP6B6 overexpression by 2-tridecanone (2-TD) induced, a new transcription factor that is forkhead box unclassified protein (Fox-uc) could bind the CYP6B6 promoter. On the basis, this project aims to confirm the Fox-uc function, and its regulation in the expression of CYP6B6 under 2-TD stress. The study will utilize RNA interference technology to identify the role of Fox-uc protein in the growth and detoxification, then through electrophoretic mobility shift assay to determine the binding sites of Fox-uc on the CYP6B6 promoter. Finally, using chromatin immunoprecipitation assay confirm Fox-uc regulates the transcription of CYP6B6 after 2-tridecanone treating. This project will further elucidate the regulatory mechanism of CYP6B6 expression and improve the metabolic pathway of cotton bollworm in response to plant secondary substance stress. It has great significance for mastering the detoxification mechanism of cytochrome P450 and providing new control target in cotton bollworm.