白纹伊蚊细胞凋亡通路及其对蚊媒病毒感染的影响

Interrupting vector competence for pathogens by genetic manipulation of mosquito vectors is a potential approach to control mosquito transmitted viral diseases. This approach requires understanding of mosquito-borne virus infection at molecular level. Apotosis plays a role in defense against virus in insects. However, apotosis pathway in mosquito is still not very well elucidated. Research on the effect of apotosis on mosquito-borne virus infection in mosquitoes at molecular level has not being reported yet. Aedes albopictus is the most harmful Aedes mosquito species in China. It is the most important vector for dengue virus in China now. Also it is resistant to all the insecticides used nowadays. Aedes albopictus mosquito is not the focus in the field of mosquito research and thus has not well studied yet. We have identified 17 apoptosis related genes of Aedes albopictus. Also, we finished the expression in E. coli, the assay of activities, the identification of the cleavage sites and active sites, the functions in cells of some of the genes indentified. This proposal plans to complete the biochemical activities and the functions in cells of all the genes identified, define the apoptosis pathway of Aedes albopictus at cell culture level, study the effect of apoptosis on mosquito-borne virus infection in cells, and finally elucidate the apoptosis pathway, the effect and molecular mechanism of apoptosis on mosquito-borne virus infection in Aedes albopictus mosquitos. The success of the work proposed above would provide new targets for the development of engineered-mosquitoes which are unable to transmit viral diseases and also would fill the gap in the research field of mosquito vector.

通过遗传操作破坏蚊子媒介效能是控制病毒性蚊媒传染病的新途径，该途径要求在分子水平理解蚊子抗病毒机制。细胞凋亡在昆虫抗病毒防御中起重要作用，目前，在个体水平研究蚊媒细胞凋亡通路及其对蚊媒病毒感染的影响仍属空白。白纹伊蚊是我国危害最大、分布最广的伊蚊蚊种，是我国目前登革热的主要传播媒介；我国境内白纹伊蚊对常用杀虫剂均产生不同程度抗性；国际上对白纹伊蚊研究很少。我们完成了17个白纹伊蚊细胞凋亡相关基因的鉴定、完成了其中部分基因的原核表达和生化活性研究、确定了其中个别caspase的切割位点和活性位点、初步观察了其中个别基因在细胞水平的功能。本项目拟在此基础上，完成白纹伊蚊细胞凋亡基因生化功能和在细胞中功能的研究；在细胞水平鉴定白纹伊蚊细胞凋亡通路；在个体水平确定白纹伊蚊细胞凋亡通路；在个体水平阐明细胞凋亡对蚊媒病毒感染的影响及其分子机制，为构建丧失媒介效能的蚊子提供新靶标，填补蚊媒研究领域空白。

蚊媒病毒是人类疾病重要病毒来源，严重威胁人类健康。白纹伊蚊在我国分布广，是重要传染病媒介，在我国引起多次传染病流行或爆发，控制白纹伊蚊对我国人民健康非常重要。细胞凋亡是细胞对病毒感染的固有反应，细胞凋亡与病毒感染机体的最终结果密切相关。病毒感染机体时，机体中被感染细胞在病毒感染早期启动细胞凋亡，可极大限制病毒复制，从而保护机体免受病毒感染导致的损伤。因此，鉴定白纹伊蚊细胞凋亡通路并研究其与病毒感染的关系，对控制白纹伊蚊及其传播的疾病具重要意义。. 本项目以白纹伊蚊为研究对象，在鉴定了白纹伊蚊17个凋亡相关基因，研究了其中部分基因表达蛋白的生化活性的前期基础上，完成了所有17个凋亡相关蛋白的原核表达及生化活性研究、确定了caspase的切割位点和活性位点；研究了其在细胞水平的功能；鉴定了白纹伊蚊细胞凋亡通路；在细胞水平、个体水平研究了其中部分蛋白与病毒感染的关系。白纹伊蚊细胞凋亡研究尚无报道，本项目研究结果有一定创新性，并为从调控细胞凋亡的角度控制白纹伊蚊及其传播的疾病提供了依据。