致倦库蚊唾液酸促进乙型脑炎病毒感染蚊虫的机制研究

乙型脑炎病毒（JEV）属于蚊媒黄病毒，主要传播媒介是库蚊属蚊虫，但是库蚊感染和传播该病毒的分子机制有待深入解析。我们前期研究发现，JEV感染可以显著上调致倦库蚊体内唾液酸合成相关的CMP-唾液酸合成酶（CSAS）和唾液酸转移酶（ST）的表达，而阻断致倦库蚊唾液酸能明显抑制JEV感染蚊虫。基于此本研究将重点研究致倦库蚊唾液酸促进JEV感染的分子机制。包括探究致倦库蚊唾液酸与JEV的相互作用，即通过qRT-PCR、IFA、ELISA、CO-IP等方法检测致倦库蚊唾液酸对JEV入侵过程的影响以及致倦库蚊唾液酸与JEV包膜蛋白的结合区域；同时探究致倦库蚊唾液酸对蚊虫免疫的潜在调控，即通过转录组学分析唾液酸对致倦库蚊相关免疫基因的调节和通过质谱鉴定致倦库蚊唾液酸修饰蛋白并探索其对JEV感染的功能。本研究为深入理解JEV感染与致倦库蚊的关系提供数据支持，也为阻断库蚊传播JEV提供理论依据。

Japanese encephalitis virus (JEV) belongs to arthropod-borne flavivirus and is principally transmitted by Culex mosquitoes, but the molecular mechanisms by which the mosquito vector participates in infection and transmission are unclear. Our previous research demonstrated that the expression of cytidine monophosphate sialic acid synthetase (CSAS) and sialyltransferase (ST) were significantly upregulated by JEV in culex quinquefasciatus. JEV infection in culex quinquefasciatus was reduced by blocking the expression of sialic acid. Therefore, we will perform a comprehensive study to explore the molecular mechanisms of how sialic acid in culex quinquefasciatus assists JEV infection. In order to investigate the interaction between sialic acid in culex quinquefasciatus and JEV, we will analyze the function of sialic acid during JEV entry and check the binding domain of JEV E protein with sialic acid by qRT-PCR、IFA ELISA and CO-IP. On the other hand, to explore the potential regulation in mosquito immune response by sialic acid, we will carry out transcriptome analysis by RNA-seq to find out the immune effectors regulated by sialic acid in culex quinquefasciatus and use mass spectrometry analysis to identify the target proteins modified by sialic acid. This study will offer further data for thorough comprehension of the relationship between JEV and Culex mosquitoes and provide new clues for blocking JEV transmission.