BmNPV与宿主家蚕中肠及胞内蛋白互作的分子机制研究

家蚕核型多角体病毒病（BmNPV）是蚕业生产上危害最为严重的一种蚕病，也是研究病毒与宿主互作的良好模型。项目组前期研究发现了病毒入侵家蚕中肠细胞过程中与胞内膜蛋白存在互作，并且在不同抗性水平的家蚕材料中表现出差异表达。本研究拟开展BmNPV 病毒与家蚕中肠细胞表面膜蛋白的互作以及入侵家蚕中肠细胞后如何利用家蚕细胞内膜系统进行体内转运的蛋白的互作过程。项目拟采用2-D电泳结合Far-western技术筛选与病毒互作的家蚕蛋白，采用BN PAGE技术鉴定病毒与宿主互作的蛋白复合体，再以转基因和RNAi在细胞验证候选蛋白的功能，同时在抗性家蚕品系和感性家蚕品系中验证候选蛋白与家蚕抗病性的可能关系。希望通过亚细胞结构蛋白互作的探索，获得NPV入侵家蚕以及细胞内转运的完整信息，寻找NPV结合的靶点，为进一步研究BmNPV防治新策略、研究媒介昆虫病原与宿主的互作、控制媒介昆虫的病原传播提供理论依据。

BmNPV is the most seriously disease of sericulture, it is also a mode of study on interaction virus with host. We have identified some interactional protein between BmNPV with silkworm midgut and these also showed difference expression in difference resistant level silkworm strains to BmNPV. Our project is aimed to study on the interaction protein between BmNPV with the membrane protein of midgut and subcellular combined protein of the subcellular membrane system. 2-D elctrophoresis combined Far-western will used to identify interaction protein, BN PAGE will used to identify interaction protein complex and transgenic gene and RNAi technologies will be used to verification the function of the candidate protein in BmN cell lines. Meanwhile, we will identify the expression level at different resistant materials to find clues of the relationship between the candidate proteins with resistant mechanism to BmNPV. Our perspective will reveal molecular mechanism of the BmNPV infection the silkworm midgut and inner transfer of midgut cell, and will find BmNPV combined targets to get a new strategy of control BmNPV diseases. The results will also benefit to support for studying on interaction pathogen between insect hosts and control the transmission of the pathogen.