褐飞虱R2D2基因的克隆及表达分析  

Cloning and Expression Analysis of R2D2 Gene of Nilaparvata lugens

作　　者：闸雯俊[1] 李三和[1] 周雷[1] 刘凯[1] 陈志军[1] 杨国才[1] 徐华山[1] 李培德[1] 游艾青[1] 

机构地区：[1]粮食作物种质创新与品种改良湖北省重点实验室/湖北省农业科学院粮食作物研究所,武汉430064

出　　处：《湖北农业科学》2016年第16期4294-4297,共4页Hubei Agricultural Sciences

基　　金：转基因生物新品种培育重大专项(2014ZX0800101B);国家青年科学基金项目(31501654);农业部华中作物有害生物综合治理重点实验室(2015ZTSJJ10);湖北省农业科技创新中心项目(2015-620-003-001)

摘　　要：R2D2蛋白广泛分布于各种生物体中,R2D2是si RNA介导的RNA干扰途径中的关键组分。R2D2含有一个串联的ds RNA结合功能域,与Dcr-2在体内形成杂合二聚体,在si RNA装载入RISC时发挥作用。本研究克隆了褐飞虱(Nilaparvata lugens St覽l)R2D2基因(NLR2D2),结果表明,NLR2D2基因全长1 673 bp,含有一个长1 005 bp的开放阅读框,编码335个氨基酸。NLR2D2蛋白与蚂蚁亲缘关系最近。利用q RT-PCR检测NLR2D2基因在褐飞虱不同发育阶段的表达,发现NLR2D2转录本在褐飞虱的所有发育阶段都有表达,在1龄若虫中表达量最低,随着生长发育表达量逐渐升高。NLR2D2基因的序列特征和表达谱非常保守,为其功能的分析提供了信息。R2D2 protein is the key component of RNAi-mediated pathway and is present in all organisms from bacteria to human. The R2D2 / Dcr-2 heterodimer is the key constituent of the RISC loading complex（RLC）. R2D2,also binds Dcr-2 and is required to load si RNAs into RISC. In this study,the Nilaparvata lugens（Hemiptera：Delphacidae） R2D2（NLR2D2） gene was dentified and characterized. The complete m RNA sequence of NLR2D2 was 1 673 bp,with an open reading frame of1 005 bp encoding a protein of 335 amino acids. The amino acid is identified with the closely related species A. echinatior.The developmental expression analysis using quantitative real-time reverse transcriptase PCR suggested that the NLR2D2 transcript was expressed in all developmental stages of N. lugens. The relative expression level of NLR2D2 was the lowest in 1st instar and increased with larval growth. These results suggest that the sequence character and expression profile of NLR2D2 were well conserved and provided the basic information for its functional analysis.

关 键 词：褐飞虱(Nilaparvata LUGENS St覽l) R2D2基因 克隆 表达 

分 类 号：Q78[生物学—分子生物学]