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Development of new defense systems in mosquito for West, Nile virus infection in Japan

日本开发出针对西尼罗河病毒感染的蚊子新防御系统

基本信息

批准号：
15580270
负责人：
MIZUTANI Tetsuya
金额：
$ 2.3万
依托单位：
National Institute of Infectious Diseases
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580270/
关键词：
siRNA Aedes albopictus West Nile virus chitinase JNK マイクロインジェクション蚊 2本鎖RNA RNAi法

项目摘要

The purpose of this research is establishment the method of controlling mosquito's growth by clarifying biological functions of JNK, and development of a new insecticide based on this fundamental research.The following were clarified from the research in 2003 and 2004.1.It was succeeded in the obstruction of growth by exposing a JNK inhibitor (SP600125) to first instar larvae of Aedes albopictus In mosquito cultured cell, JNK was confirmed to have a function of anti-apoptosis.2.JNK was observed the activation by adding Lipopolysaccharide(LPS) to mosquito cells, and a novel gene, which should code anti-bacterial peptide, was discovered from the cells.3.The transfection method of siRNA was examined for evaluating the function of JNK. Although inicroinjection siRNA against JNK to third instar larvae lead to decrease growth of mosquito, it was impossible to handle microinjection method to a lot of larvas. Then, we tried to improve the method of siRNA using reagents for transfection. However, we could not obtain superior to the microinjection method.4.First instar larvae was bred in the culture medium containing potent insect chitinase inhibitors of fungal origin, and the growth was obstructed by two kinds of culture medium. The gene that codes the inhibitors is now under identification.In this research, we clarified that JNK plays important roles in mosquito. Moreover, potent inhibitors of chitinase are useful for inhibition growth of mosquito.

本研究的目的是通过阐明JNK的生物学功能，2003年和2004年的研究结果表明：1.通过暴露JNK抑制剂，成功地抑制了植物的生长（SP600125）对白纹伊蚊（Aedesalbopictus）1龄幼虫的抑制作用，证实JNK在蚊培养细胞中具有抗细胞凋亡的作用：2.观察了脂多糖（Lipopolysaccharide，LPS）对蚊细胞JNK的激活作用，发现一个编码抗菌肽的新基因，3.探讨siRNA转染方法对JNK功能的影响。虽然针对JNK的siRNA对3龄幼虫的显微注射可以抑制蚊虫的生长，但不可能对大量幼虫进行显微注射。在此基础上，我们尝试改进siRNA转染试剂的制备方法。但不能获得比显微注射法更上级的效果。4.用含有真菌来源的几丁质酶抑制剂的培养基培养1龄幼虫，两种培养基均能抑制1龄幼虫的生长。编码JNK抑制剂的基因正在鉴定中，本研究阐明了JNK在蚊虫中的重要作用。此外，有效的几丁质酶抑制剂可用于抑制蚊子的生长。