柞蚕核型多角体病毒基因组编码VmiRNA的预测与功能分析  

Prediction and Functional Analysis of VmiRNA Encoded by Genome of Antheraea pernyi Nucleopolyhedrovirus

作　　者：李文利[1] 林澜[2] 王乃红 栾雨时[1] 

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]沈阳药科大学生命科学与生物制药学院,沈阳110016 [3]山西省蚕业科学研究院,山西运城044000

出　　处：《蚕业科学》2013年第2期324-332,共9页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31171971)

摘　　要：柞蚕核型多角体病毒(ApNPV)是危害柞蚕的主要病原物之一。病毒microRNAs(VmiRNA)是由病毒基因组编码的小RNA分子,在病毒繁殖及与宿主的相互作用中发挥作用。采用高通量测序Solexa技术测定ApNPV感染柞蚕后的VmiRNA,并通过vMir软件预测ApNPV基因组可能编码的VmiRNA前体序列。在预测得到的16条前体序列中,有13条能够从病毒感染的柞蚕蛹脂肪体中扩增出条带,并有10条能在病毒基因组中找到同源序列。在ApNPV MicroRNA Solexa测序数据库中搜索到与这10条前体VmiRNAs相匹配的序列,用RT-PCR方法证明有9条能扩增出大小相应的片段,确定其为病毒基因组编码的VmiRNA的成熟序列。利用靶基因在线预测程序RNAhybrid预测到这些VmiRNA作用的135个可能的靶基因,其中有与免疫应答过程相关的基因36个,与免疫系统进程相关的基因19个,与免疫调节相关的基因8个,与免疫系统发育相关的基因9个。推测这些ApNPV基因组编码的VmiRNA在病毒与宿主之间的相互作用中扮演重要角色。Antheraea pernyi nucleopolyhedrovirus(ApNPV) is one of the major pathogens infectious to Antheraea pernyi.Viral VmiRNAs are small RNA molecules encoded by virus genome,which play important roles in virus replication and virus-host interaction.In this study,the solexa high-throughput sequencing technology was employed to determine VmiRNAs in Antheraea pernyi being infected by ApNPV and vMir software was used to predict potential VmiRNA precursor sequences encoded by ApNPV genome.Among the 16 predicted precursor sequences,13 could be amplified from Antheraea pernyi pupal fat body with ApNPV infection,and 10 of them had homologous sequences in the viral genome.Sequences matching these 10 precursor VmiRNAs were obtained from searches in the database of solexa sequencing result,and 9 of them were verified by RT-PCR to yield fragments of correspondent sizes,displaying that they were mature VmiRNA sequences encoded by ApNPV genome.These mature VmiRNAs were predicted to have 135 potential target genes by online RNAhybrid software,among which 36 genes were involved in immune response and process,19 genes in immune system process,8 genes in immune regulation,and 9 genes in immune system development.These data indicate that ApNPV VmiRNAs play important roles in virus-host interaction.

关 键 词：柞蚕核型多角体病毒 Solexa测序 VmiRNA 靶基因 

分 类 号：S885.1[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]