家蚕核型多角体病毒lef-11基因对病毒增殖的调控研究

项目名称： 家蚕核型多角体病毒lef-11基因对病毒增殖的调控研究

项目编号： No.31472152

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 畜牧学与草地科学

项目作者： 潘敏慧

作者单位： 西南大学

项目金额： 86万元

中文摘要： 家蚕是重要经济昆虫, 同时也是鳞翅目模式昆虫。家蚕核型多角体病毒是家蚕重要病害家蚕核型多角体病的病原，由于其侵染增殖的分子机制未完全解析，阻碍了家蚕抗BmNPV病毒的抗性机制解析和家蚕抗病毒育种进程。本项目在前期研究中通过对BmNPV编码基因进行逐一沉默，筛选到了病毒复制增殖的关键基因lef-11。本项目拟采用分子生物学和生物信息学等方法，鉴定LEF-11核定位信号，揭示其核转运机理；构建lef-11基因敲除病毒，解析其对病毒增殖的分子调控机制；通过免疫共沉淀技术筛选并鉴定与LEF-11互作的宿主蛋白，分析过表达lef-11的家蚕细胞中的差异表达基因，阐明LEF-11调控宿主细胞的分子机理。通过本项目研究，可完善BmNPV侵染增殖的分子机制和揭示BmNPV与家蚕的互作机制；可为解析家蚕抗BmNPV病毒的抗性机制、加快家蚕抗病毒育种进程以及开发高效的生防制剂奠定基础。

中文关键词： 基因功能；转录调控；分子机制

英文摘要： Silkworm is an important economic insect, but also the mode insect of Lepidoptera. Bombyx mori nuclear polyhedrosis virus（BmNPV） is pathogen of the Bombyx mori nuclear polyhedrosis disease. Because the molecular mechanisms of BmNPV infection and proliferation are not fully resolved, the anti-viral mechanisms resolving and antiviral breeding process of silkworm were hampered. In the early studies this project, more than 100 genes of BmNPV were silent one by one by RNAi tech, the gene silencing effects on proliferation and replication of BmNPV were compared under these same conditions, the lef -11,proliferation and replication key gene of BmNPV,were selected. We will use molecular biology and bioinformatics methods, idenfify nuclear localization signal and reveal the nuclear transport mechanism of LEF-11;analyze viral DNA replication and viral gene expression regulation, reveal the regulating molecular mechanisms of the virus proliferation;screen and identify the interaction proteins between the LEF-11and host cells,detect the transcriptome and proteome changes in BmN-SWU1 cells with lef-11 gene overexpression;establish the LEF-11 protein regulatory networks and elucidate the molecular mechanism of LEF-11 regulation of host cells. The project may lay the foundation for revealing the anti-viral mechanisms and antiviral breeding process of silkworm, improving the efficiency of baculovirus expression system and developing effective biocontrol agents.

英文关键词： gene function;transcriptional regulation;molecular mechanisma