柞蚕免疫系统相关基因ApTOLL1的克隆及两种蚕微孢子虫感染下的诱导表达

目的]鉴定柞蚕的免疫相关TOLL样受体家族基因,以期为进一步探讨柞蚕的免疫机制奠定基础。方法]克隆了柞蚕免疫系统相关TOLL样受体家族基因,并进行序列和系统进化分析,同时,利用两种蚕微孢子虫侵染柞蚕,检测TOLL样受体相关基因的表达差异,分析不同病原微孢子虫感染柞蚕后所引起的免疫应激差异。结果]通过cDNA克隆测序获得一个可能与柞蚕免疫相关的基因片段,序列及系统进化分析显示它与家蚕Toll信号通路中的Toll1基因最为同源,将其命名ApTOLL1基因。进一步对柞蚕蛹分别注射家蚕微孢子虫和柞蚕微孢子虫后,通过实时荧光定量PCR技术分析显示,注射家蚕微孢子虫2 h后蚕蛹内ApTOLL1大量表达,而注射柞蚕微孢子虫11 h后蛹内该基因才开始表达,这表明柞蚕Toll信号通路针对不同病原微孢子所诱导产生的免疫响应时间有所差异。结论]该研究结果首次克隆了柞蚕ApTOLL1基因,并为进一步研究柞蚕的免疫机制提供了帮助。

Cloning of Immune System-related Gene ApTOLL1 and Its Expression in Nosema-infected Antheraea pernyi

Objective] This study aimed to identify the immune system-related TOLL-like receptor family gene of Antherea pernyi,to lay the foundation for further investigating the immune mechanism of Antherea pernyi.Method] Immune system-related TOLL-like receptor family gene of Antherea pernyi was cloned for sequencing and phylogenetic analysis;in addition,expression variations of TOLL-like receptor family gene in Antherea pernyi infected with Nosema pernyi and Nosema bombycis were detected to analyze the differences in immunological reactivity of Antherea pernyi to Nosema infection.Result] Based on cDNA cloning and sequencing,an immune system-related gene fragment was obtained from Antherea pernyi,which is the most homologous to the Toll1 gene in Toll signaling pathway of Bombyx mori according to the sequencing result and phylogenetic analysis,and is named ApTOLL1 gene.Subsequently,Antherea pernyi pupae were injected respectively with Nosema bombycis and Nosema pernyi,fluorescent real-time quantitative PCR analysis showed that ApTOLL1 was abundantly expressed in Antherea pernyi pupa 2 h after Nosema bombycis injection,which began to express in Antherea pernyi pupa 11 h after Nosema pernyi injection,indicating that the immune response time induced by various Nosema strains varies in Toll signaling pathway of Antherea pernyi.Conclusion] ApTOLL1 gene of Antherea pernyi was first cloned in this study,which provides reference for further investigating the immune mechanism of Antherea pernyi.