基于mRNA测序的家蚕中肠免疫转录组分析

项目名称： 基于mRNA测序的家蚕中肠免疫转录组分析

项目编号： No.31272497

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 吕志强

作者单位： 西北农林科技大学

项目金额： 80万元

中文摘要： 肠道作为阻止病原物入侵的第一道防线在昆虫天生免疫中起着重要的作用。以果蝇为模式，利用分子遗传学手段研究发现活性氧反应和通过IMD途径产生抗菌肽是肠道中的主要防御反应机制。但对于其它昆虫的肠道免疫机制，则所知很少。对于是否存在除活性氧和抗菌肽之外的其它免疫机制也不清楚。本项目以家蚕为研究对象，利用RNA-Seq技术，分析家蚕在不同类型细菌感染后的中肠转录组变化，鉴定差异表达基因，利用qRT-PCR对抗菌肽诱导信号途径的关键基因进行验证，测定活性氧反应机制的关键酶活性，从而明确活性氧和抗菌肽在家蚕肠道免疫中的作用，揭示家蚕肠道防御反应的机制，并确立与果蝇肠道免疫机制的进化关系，为肠道免疫系统关键分子的功能分析及机理研究提供线索。

中文关键词： 家蚕；肠道；细菌；免疫；RNA-Seq

英文摘要： Intestine plays critical roles in insect immune systems as the front line to confront the invading pathogens. Using fruit fly Drosophila melanogaster as a model, researchers have found that the generation of reactive oxygen species (ROS) and the production of antimicrobial peptides (AMP) through IMD pathway are the main mechanisms in gut immune responses. Little is known about the gut immunity in other insects than fruit fly. It is also unclear so far whether other responses contribute to the defense except AMP and ROS. Taking the advantage of RNA sequencing, we try to analyze the transcriptome of silkworm Bombyx mori midgut after oral infection by feeding of different pathogenic bacteria to identify the differential expressed genes. We will verify the key genes in AMP regulation pathways by quantitative RT-PCR and measure the enzyme activity involved in ROS metabolism. This project will not only define the functions of ROS and AMP in the silkworm gut defense, also will reveal currently unknown pathways or mechanisms in gut immune responses. The silkworm gut immune transcriptome will provide phylogenetic comparison of gut defense mechanisms between silkworm and fruit fly. It will give us some clues for the functional study of key molecules in the insect intestine immunity as well.

英文关键词： Bombyx mori；intestine；bacteria；immunity；RNA-Seq