家蚕中肠特异启动子的构建及在抗病毒遗传改良中的应用

项目名称： 家蚕中肠特异启动子的构建及在抗病毒遗传改良中的应用

项目编号： No.31501875

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 动物学

项目作者： 蒋亮

作者单位： 西南大学

项目金额： 20万元

中文摘要： 家蚕质型多角体病毒（BmCPV）和家蚕浓核病毒（BmDNV）只侵染家蚕中肠，给蚕业生产造成了严重经济损失。由于有关BmCPV和BmDNV侵染家蚕的机制以及蚕体的免疫防御机制都不甚清楚，干涉病毒基因是目前抑制这两种病毒最有效的策略。我们前期研究克隆了家蚕中肠特异表达基因BmAPN4的启动子序列，命名为P3启动子（P3P），检测结果显示BmAPN4在家蚕整个中肠表达而P3P只在中肠前部具有活性。生物信息学分析发现在P3P和BmAPN4的翻译起始位点ATG之间含有一个5′UTR内含子（5UI），推测其可能影响了BmAPN4的表达。本项目将克隆该5UI，对其功能进行研究并构建在家蚕整个中肠都有活性的特异启动子，进而利用该启动子介导BmCPV多个基因的串联干涉及BmDNV多个基因的串联干涉，制备抗BmCPV和抗BmDNV的转基因家蚕品系。本课题的实施将在基础研究和应用研究两方面都具有重要参考价值。

中文关键词： 功能基因组；生物信息；基因结构与调控；转基因昆虫；RNA干扰

英文摘要： BmCPV and BmDNV only infect the midgut of silkworms, which cause severe economic losses to sericulture. The infection mechanisms of BmCPV and BmDNV and the immune defense mechanisms of host are unclear, RNAi of viral genes are the most effective strategy to inhibit the two viruses. BmAPN4 is a midgut-specific gene in silkworm, of which the promoter sequence was cloned and named the P3 promoter (P3P) in our previous study. Detection results showed that BmAPN4 was expressed in the entire midgut while P3P only had activity in the anterior midgut. Bioinformatics analysis revealed that there was a 5′UTR intron (5UI) between P3P and the translation start site ATG of BmAPN4, this 5UI may affect the expression of BmAPN4. In this project, we will clone the 5UI of BmAPN4 and study its function, construction of a specific promoter with activity in the entire silkworm midgut. And then, the dsRNA for multiple tandem genes of BmCPV would be controlled by this specific promoter to generate transgenic silkworms with resistance to BmCPV, and anti-BmDNV silkworms would be also generated by RNAi of viral multiple genes using this promoter. In conclusion, the implementation of this project will have an important reference value in basic research and applied research.

英文关键词： functional genomics;bioinformatics;gene structure and regulation;transgenic insects;RNAi