柞蚕核型多角体病毒内源性非编码小RNA成熟体MD5作用的宿主靶基因及功能预测  

Target-gene Prediction and Functional Analysis of Endogenous Non-coding MicroRNA MD5 from Antheraea pernyi Nucleopolyhedrovirus

作　　者：宋丽新[1] 刘丹梅[2] 李文利[3] 

机构地区：[1]大连理工大学生命科学与技术学院,辽宁大连116024 [2]辽东学院农学院,辽宁丹东118001 [3]大连理工大学生命与医药学院,辽宁盘锦124221

出　　处：《蚕业科学》2015年第1期72-79,共8页ACTA SERICOLOGICA SINICA

基　　金：瑞典研究理事会项目

摘　　要：研究病毒内源性非编码小RNA(VmiRNA)的靶标基因,有助于阐释病毒和宿主的互作关系及制定对病毒的防控策略。利用Solexa高通量测序技术及生物信息学方法,从柞蚕核型多角体病毒(Ap NPV)基因组中筛选出VmiRNA成熟体MD5,在预测其作用的宿主靶基因基础上,通过GO数据库分析靶基因的功能。结果显示由Ap NPV的VmiRNA MD5调控的宿主靶基因涉及细胞组成、分子功能、生物学过程等生物学途径,主要参与细胞内的分子结合、酶催化和免疫反应,其中与柞蚕免疫相关的靶基因共有4个。实时荧光定量PCR检测柞蚕蛹经Ap NPV诱导处理后VmiRNA MD5在脂肪体内明显上调表达(与对照相比提高了8倍),而与宿主免疫相关的3个靶基因的表达水平明显下调。对VmiRNA MD5 5'端上游启动子序列结构的分析显示,其含有免疫相关转录因子GMCSF、p53等的结合位点。推测VmiRNA MD5可能参与调控宿主柞蚕免疫相关基因的表达。To study the target genes of viral endogenous non-coding microRNA（ VmiRNA） will help understand the interactive relationship between viruses and their hosts and formulate prevention and control strategy against virus. In this paper,a VmiRNA mature sequence MD5 was screened from Antheraea pernyi nucleopolyhedrovirus（ Ap NPV） genome by using Solexa high-throughput sequencing techniques combined with bioinformatic method. Based on target gene prediction,GO analysis was performed to analyze functions of the target genes. The result showed that the target genes of VmiRNA MD5 are mainly involved in cell composition,molecular function,and biological process pathway,and are related to molecular binding,catalytic and immune system,in which there are 4 immunity related genes from Antheraea pernyi. Real-time quantitative PCR detection indicated that the expression level of VmiRNA MD5 was obviously upregulated in pupal fat body of A. pernyi after infection by Ap NPV（ increased by 8 folds compared to the control group）,while 3 target genes related to host immunity were obviously down-regulated. Structural analysis of promoter elements showed that 5＇-terminal upstream sequence of VmiRNA MD5 contains several binding sitesfor transcription factors such as GMCSF and p53,suggesting that MD5 is probably involved in regulation of immune-related gene expression in A. pernyi.

关 键 词：柞蚕核型多角体病毒 内源性非编码小RNA 生物信息学 靶基因 启动子 

分 类 号：S884.51[农业科学—特种经济动物饲养]